65 2006 PDF

PL ISSN 1506-9427
Vegetable Crops Research Bulletin

Vol. 65/2006
RESEARCH INSTITUTE OF VEGETABLE CROPS INSTYTUT WARZYWNICTWA SKIERNIEWICE, POLAND
Vegetable Crops Research Bulletin starting from vol. 49/1998 replaces Biuletyn Warzywniczy Bulletin of Vegetable Crops Research Work founded in 1953
Editorial Board A. Dobrzaski (Editor - in - Chief), F. Adamicki (Associate Editor) Editors: R. Kosson, J. Szwejda, K. Grecka, Managing Editor: B. Nowak
Editorial Advisory Board S. Kaniszewski (Poland) - Chairman I. Babik (Poland), C. Bobinas (Lithuania), J. Czapski (Poland), J. Dyduch (Poland), K. Elkner (Poland), M. Horbowicz (Poland), M. Knaflewski (Poland), E. Koota (Poland), A. Libik (Poland), K. Niemirowicz-Szczytt (Poland), D. OConnor (United Kingdom), A. Polyakov (Russia), R.K. Prange (Canada), S. Pruszyski (Poland), J. Robak (Poland), H. Skpski (Poland), K. Szudyga (Poland), F. Tei (Italy), M. Valikov (Slovak Republic), C.L.M. de Visser (Netherlands)
Partially subsidised by: POLISH SOCIETY FOR HORTICULTURAL SCIENCE MINISTRY OF EDUCATION AND SCIENCE POLISH ACADEMY OF SCIENCES
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Editorial Office Research Institute of Vegetable Crops (RIVC) Instytut Warzywnictwa 96-100 Skierniewice, Konstytucji 3 Maja 1/3, Poland phone: +48(46) 833 22 11, fax: +48(46) 833 31 86 www.inwarz.skierniewice.pl e-mail: bulletin@inwarz.skierniewice.pl
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Instructions for authors of publications in Vegetable Crops Research Bulletin

The journal, Vegetable Crops Research Bulletin covers fundamental and applied research in vegetable crops and publishes original papers as well as reviews and reports from symposia and conferences in English. Submission of a manuscript implies that the paper has not been published or submitted for publication elsewhere. All papers which have been qualified as relevant with the scope of our journal are reviewed. The main text should not exceed 12 pages of normalized type script (A-4 size) including illustrations and tables (not exceeding the A-4 size). Each table and figure should be placed on separate page. Original prints (obtained from inkjet or laser printer) on white paper should be provided. It is important that lines an symbols should be large enough to enable reduction to the desired size. Paper prepared in computer text editor Word for Windows should be delivered on diskette. The diskette must be accompanied by copy of computer printout. Label the diskette with your name, the file name and the program used. Papers should be sent to the address: Vegetable Crops Research Bulletin Research Institute of Vegetable Crops 96-100 Skierniewice, Konstytucji 3 Maja 1/3, Poland tel. +48 (046) 833 22 11, fax: +48 (046) 833 31 86 e-mail: bulletin@inwarz.skierniewice.pl The name(s) of author(s) in full and the name of institution where the work was done should be written below the title. Every paper should be divided under the following headings in this order: Summary, Introduction, Materials and Methods, Results, Discussion, Conclusions, References. The key words (up to 4-6 words) should follow the Summary. Metric SI units should be used in paper. All Latin names should be written in italics, or text fragment to be printed in italics should be clearly indicated on the paper copy. References should include the names of authors in alphabetical order according to example: Author A., Author B. 1999. Title of article. Journal Title 20: 35-42. Quoted Russian papers should be transliterated according to the standard international system ISO. The abbreviated titles of periodicals according to Word list of scientific periodicals should be used in References. The periodicals titles not included in this list should be written in full name as a not abbreviated. When book is quoted the title of book, editors and place of publication should be given. Author A., Author B. 2000. Chapter Title. pp. 21-47. In: Book Title (A.B. Editor ed.) Publisher, Place, Country. Author A., Author B., Author C. 2000. Book Title (A.B. Editor) Publisher, Place. The Polish Summary (up to 300 words) should be added after References. Summary for foreign authors will be translated into Polish by the editor. The papers are published in the order of acceptation for publication. The payment for publication is free of charge, till now. The authors will receive 10 offprints free of charge.
From Editor The Vegetable Crop Research Bulletin publishes mainly research papers and occasionally reports from conferences on topics of science and technology related to vegetable crops. The current issue presents research papers and Proceedings of the Third International Onion Network Meeting Accelerating technology transfer in East European onion production organised by Prof. Franciszek Adamicki, Research Institute of Vegetable Crops, Skierniewice, Poland and Prof. Grigoriy I. Yarovoy - Director of Institute of Vegetables and Melons, Ukrainian Academy of Agricultural Science, Kharkov, Ukraine and held on July 25-26 July in Kharkov (Ukraine). The International Onion Network was established with the aim of collecting and disseminating information and experience with technology, plant protection, storage and marketing of onions, identifying gaps in knowledge and defining new research projects. This International Network is financed by the Polish Ministry of Education and Science. On behalf of the Editorial Board I would like to thank all contributors of reports presented in this volume. I greatly appreciate the help of organising committee for the International Onion Network in preparing the Proceedings for publication I hope that the research papers and Proceedings presented in this volume will prove useful and advance the knowledge in science and vegetable crops technology. Editor-in-Chief A. Dobrzaski
2006 VEGETABLE CROPS RESEARCH BULLETIN 65

RESEARCH INSTITUTE OF VEGETABLE CROPS SKIERNIEWICE ________________________________________________________________________________________
FUSARIUM BASAL ROT IN THE NETHERLANDS

Chris de VISSER, Rob van den BROEK, Lubbert van den BRINK Applied Plant Research Lelystad, Wageningen UR PO Box 430 NL-8200 AK Lelystad, The Netherlands
Summary Fusarium basal rot is an increasing threat to onion production in temperate regions. Research in The Netherlands made clear that tolerant varieties are available. However the yields of these varieties are still low and they are not suitable for long storage. Tolerant varieties can only be part of a total disease control strategy. Other potential tools are: avoidance of fields with basal rot incidence, destruction of infected plants, crop rotation, farm hygiene, planting materials and possibly organic soil amendments. To manage Fusarium basal rot it is needed to optimize the disease control strategy. This is only possible when a reliable and cost-effective detection procedure for soil samples and plant samples becomes available. key words: onion, Fusarium oxysporum, Sclerotium cepivorum, onion diseases INTRODUCTION In The Netherlands Fusarium basal rot is a steadily increasing problem. Fusarium basal rot is caused by Fusarium oxysporum f. sp. cepa. The disease is common in countries with warmer climates, like the USA, France and Italy. The last few decades the disease has extended into more temperate regions. This may be caused by the global warming, but also mutatation of the fungus could be a reason. The financial loss for Dutch farmers confronted with Fusarium basal rot is substantial, in the first place because of the yield reduction and the higher storage costs. In the second place because of the fact that the disease level can ultimately result in fields unsuitable for onion production. In this article the disease development will be described and some results of the research done in The Netherlands will be shown. Disease development Fusarium basal rot can attack onions at different stages of growth. Roots can be infected already in a young stage leading to early plant death. Infections in later growth stages will result in basal rot, either at harvest or in store. Infection in the field can be recognized by wilting of the onion plant following attack of the root system. This symptom becomes visible earlier under dry conditions.
VEGETABLE CROPS RESEARCH BULLETIN 65 6 _____________________________________________________________________________________________________
Early infected plants are dying. Plants which are infected later show wilting symptoms and dying leafs. In the bulbing stage the basal plate is showing discoloration and rot is extending into the bulb. Under wet conditions the fungus can produce white mycelium on the basal plate. This symptom can easily be confused with onion white rot, caused by Sclerotium cepivorum. However the difference is that with an attack of white rot sclerotia must be found in the soil and on the diseased bulbs in storage under wet conditions. Figure 1 shows wilt symptoms in the field, while Figure 2 shows bulbs with fusarium basal rot symptoms.
Fig. 1. A field with Fusarium basal rot
Fig. 2. Onion bulbs with Fusarium basal rot symptoms
Ch. de VISSER et al. FUSARIUM BASAL ROT ... 7 _____________________________________________________________________________________________________
The severity of infection is to a large degree dependent on the time of infection and plant stress factors. Under stress conditions plants are highly susceptible. This was shown in a pot trial in 1995 in which seeds, transplants and sets were sown or planted in artificial infected soil. Figure 3 shows that plants that are weakened and growing under stress conditions (transplants) were highly susceptible whereas onions from sets showed no wilt at all.
60 50
% wilt or rot
not infected low infected high infected
29.2 48.3
40 30 20 10 0
2.3
13.7 14.4
seeds
transplants
sets
infection level of the soil
Fig. 3. Influence of plant condition on infection
Also plants without symptoms but with infection can show a retarded growth. In a pot trial in 1995 it was shown that the fresh weight of healthy plants grown on infected soil was 10-30% lower in comparison with healthy plants grown on not-infected soil. Life cycle of Fusarium basal rot Fusarium basal rot is a soil borne disease. The fungus can survive in the soil either in the form of mycelium on organic matter in the soil or as resting spores called chlamydospores and microconidia. The fungus can survive in the soil in absence of the onion plant, but it has to compete with other fungi. The duration of survival is not known. Blok & Bollen (1996) found that Fusarium oxysporum can survive asparagus-free periods for at least 20 years in The Netherlands. Fusarium basal rot can easily spread with planting material on which soil is attached or by not cleaned agricultural machines. In literature it is known that the fungus can be detected in the seed but it is not known if spreading by seed is significant for current day practical conditions. The optimum temperature for growth is rather high: between 24 and 27C. However in The Netherlands Fusarium basal rot damage was also visible in mid June when soil temperature had not reached this optimum temperature range. During storage Fusarium basal rot is not spread, unless the bulbs are damaged.
Tolerant varieties Worldwide tolerance in onion varieties is considered the best way to fight Fusarium basal rot. In The Netherlands the tolerance of many varieties have been investigated. The tolerant varieties Daytona, Sundance, Takstar and Takmark were suffering less from basal rot, but the yields of these varieties were still low. Moreover, under Dutch conditions Takmark and Takstar are not suitable for long storage. The tolerance level is very variable from field to field. This can be caused by variability in the infection process, including factors such as soil and weather conditions and Fusarium oxysporum population density and virulence. The last reason seems to be important in The Netherlands. In Table 1 an example of differences in tolerance levels between varieties is shown.
Table 1. Results of cultivar testing on an infected field in the Flevopolder area in 2003 Cultivar Summit Spirit Takmark Recorra Mundial Yield (tha-1) 4 21 30 40 28 % plant loss on the field 89 58 17 26 32 % basal rot at harvest 63 30 10 11 12 % rot at harvest + in store 82 46 23 25 27
The difference between tolerant and susceptible varieties is not caused by differences in infection. Both are infected, but in tolerant varieties the fungus does not have the capacity to grow beyond the basal plate and enter the fleshy bulb material. In The Netherlands we think that tolerant varieties can only be part of a total disease control strategy. Tolerant varieties loose probably some yield potential on infested soil and also with tolerant varieties some rot still occurs at harvest and during storage. In 2002 and 2003 Applied Plant Research has investigated if there is a difference between tolerant and non-tolerant varieties in stimulating the disease. Soil has been sampled before and after growth of several varieties in a field experiment. The sampled soil was tested in a bio-assay with onion seeds. In Table 2 the results are shown.
Table 2. Bio-assay results on infection level increase after onion growing Cultivar Summit Spirit Takmark Recorra Mundial Bio-assay result Before sowing After harvest 22 31 26 30 20 22 17 25 22 29 % increase 41 15 10 47 32
It appeared that there are differences between the tolerant varieties: for example the increase in the infection level is much lower with Takmark than with Recorra or Mundial. This indicates that the resistance level of Takmark is higher. It must be stated however that the results were highly variable and that the bio-assay does not have a high level of reliability. Bio-assay In The Netherlands efforts were made to develop a bio-assay. The objective was to develop a reliable and cost-effective bio-assay to determine the disease potential of commercial production fields. It was tested if bio-assays with onion transplants, sets or seeds were giving the best results. Bio-assays with seeds were the most promising. However the reliability of this bio-assay was not good enough to predict Fusarium basal rot to occur. In Figure 4 the relation between the bio-assay result and disease severity in the field is shown.
100
Fusarium at harvest (%)
90 80 70 60 50 40 30 20 10 0 0 10 20 30 40 50 60 70 80 90 100
Bio-assay result (%)

Fig. 4. Established relation between bio-assay result and Fusarium incidence until harvest (continuous line) and the 95% confidence limits (dotted lines).
Disease prevention and control As stated above tolerant varieties can only be part of a total disease control strategy. There are different other methods to manage the disease: Avoidance of fields with a recent basal rot incidence. As a reliable and costeffective detection procedure is not yet available, it is not possible to analyze soil samples in order to determine an infection level. This leaves past onion growing history as the only reliable method to determine whether or not disease could be expected. If infected plants are found, the destruction of these plants together with healthy looking plants surrounding them could limit dissemination of the fungus through the field as mentioned by Pataky (1988) in a Fusarium control
manual for ornamentals. The removal of these plants could slow down disease development in the field. Steam sterilizing the soil and realizing a temperature of over 80oC for at least 30 minutes, or 70C for more than 1 hour (Pataky 1988). This procedure is expensive and not cost-effective for onion production. In combination with other diseases to control, this could result in some control of Fusarium basal rot. However, no data on the effect on Fusarium basal rot have been found. Chemical control with fungicides. Tebuconazole gives a reduction of fusarium when the soil is treated before onions are sown. This treatment is not allowed in the Netherlands. In China, carbendazim, thiophanate and thiophanate-methyl are fungicides that have been reported to control Fusarium wilt on watermelons. Not growing host plants for a certain number of years. The question is to what extent this control measure is effective in the case of Fusarium basal rot. Fusarium oxysporum species can mostly survive in soil by chlamydospores in plant debris (Nelson 1981).These are thick-walled spores that are assumed to be able to survive for many years, although no data were found to give insight into the survival rate of chlamydospores of Fusarium oxysporum. Furthermore, the fungus could also be able to survive on plant debris as mycelium. Data are lacking to indicate the required number of non-host years that are effective for the population level to decline to sufficiently low numbers. A rough indication in practice is a period of 3-4 years but no data were found to back up this advice. In The Netherlands we have done research on soils with a Fusarium basal rot history of 0-8 years old. The bio-assay earlier was used to find an indication of disease level. Figure 5 shows that the results were variable and no disease decline could be detected.
Fig. 5. The effect of the number of years without onion growing (X axis) on the % diseased onions plants in the bio-assay as caused by Fusarium infection (Y axis) in 2003
Different management tools can be identified for prevention of disease spread, namely: - The best possible prevention of soil infection by Fusarium oxysporum f.sp. cepae would be to only use planting materials that are free from inoculum. It is likely that the disease can spread by attachment to or infection of planting material such as sets. In the Netherlands, a system is in place with NAK-T where sets are certified free from disease. Sets from fields with disease incidence must not be used. However, a detection procedure on sets to ascertain the absence of pathogenic Fusarium (for instance PCR-based genetic techniques) could further limit the risk of disease spread. Moreover, it cannot be ruled out that the disease could spread with other planting material as well as onion sets. Theoretically, soil attached to potato seed, tulip bulbs or other planting material could also produce a potential risk. - In years with very low onion prices it can happen that produce cannot be sold. This could result in higher volumes of onion waste. It would be wise to prevent this waste from ending up on non-infected soils. - Furthermore, prevention of disease spread can be based on the principle of good farm hygiene. The fungus can spread with soil attached to machinery and tools, boots etc. when moving from an infected field to a not infected one (Mace et al. 1981).
- It is known that some Fusarium oxysporum fungi can spread with irrigation water (Kommedahl et al. 1970). In the case of Fusarium oxysporum f. sp. cepae no records can be found to support this disease dissemination, but it cannot be ruled out. Soil solarization alone or in combination with fumigants. In countries with a warm summer climate this is effective in controlling pathogenic soil fungi. In France, soil solarization has not proven to be effective enough for completely eliminating the inoculum from soil in the case of Fusarium oxysporum f. sp. dianthi. The addition of small quantities of fumigants (metham sodium or methyl bromide) significantly improves its efficacy. Satisfactory crop growth up to the second year of culture was possible (Cebolla et al. 1993). However, this method is not cost-effective for onion growing and not applicable in temperate regions. Organic soil amendments could possibly be effective in controlling diseases caused by Fusarium oxysporum. Amendments such as mustard (Brassica juncea) residues, rice husks, oyster shell powder, urea, KNO3, Ca superphosphate, mineral ash, wheat straw, clover straw, onion residues, coffee hulls, oilcakes from groundnut, mustard, sesame, cotton seeds, stalks of sunflower, alfalfa and Hungarian vetch (Vicia pannonica) were found very effective for the control of Fusarium wilt disease of bean, celery, melon, peas, radish and tomato caused by Fusarium oxysporum. In Figure 6 the effect of sunflower, alfalfa and vetch on basal rot in a crop following soil amendment, are shown (Ozer et al. 2002).
Fig. 6. Onion bulb rot incidence (%) in the soils amended with stalks of different plants in two experimental areas. Columns with the same letter do not differ significantly (P=0.05) (figure copied from Ozer et al. 2002).
The phenomenon of soil suppresiveness regarding Fusarium wilt in several crop species has been known for 70 years. Knowledge of the underlying principle could open perspectives for fusarium wilt control. In some soils the disease did not occur although Fusarium oxysporum f.sp. cepae was present together with a susceptible host and suitable climatic conditions. In France, research has pointed out that non-pathogenic Fusarium oxysporum isolates play an important role in the mechanism underlying soil suppresiveness. Suppressive soils are low in iron availability (Alabouvette et al. 1993). In The Netherlands, Blok et al. (1997) have shown that the pre-colonization of soil with non-pathogenic isolates of Fusarium oxysporum reduced root rot severity of asparagus plants by more than 50%. Although soil suppresiveness has been the basis of much research, it has so far not produced practically usable control measures. However, it seems promising for improvement of understanding the phenomenon as it can extend our knowledge on biological and nonbiological interactions in disease outbreaks.
Detection of Fusarium oxysporum f.sp. cepae In developing effective management strategies, prevention is a key issue. To prevent infestation or unexpected crop loss due to basal rot, it is essential to have a practical and cost efficient detection procedure. The most commonly used method is the detection of F. oxysporum with the aid of selective Komada medium (Komada 1975). This method can be used in combination with a statistical technique called soil dilution plate count. On Komada medium only F. oxysporum is able to grow at a significant rate. However, also F. avenaceum can form colonies, but this fungus grows more slowly on this medium and can easily be recognized by its appearance. Although the Komada medium is useful to determine the number of colony forming units of F. oxysporum in the soil, the result has no direct bearing on soil infestation with F. oxysporum f.sp. cepae. If this selective method is used on soil with a known history of basal rot in onions, it can give an indication of the degree of infestation (Abawi & Lorbeer 1971a). However, if the objective is to determine the infestation level on an arbitrary soil, false interpretations are likely, because the pathogenicity of a colony is not determining its growth pattern on a Komada or other selective medium. Until now an effective and reliable pathogenity test for isolates of Fusarium oxysporum f.sp. cepae is not available. Much work has been done on developing bio-assays. A bio-assay using seeds or sets has not been developed until now. It is doubtful if bio-assays with transplants are useful. In our research we have found that some isolates of F. avenaceum were pathogenic using onion transplants but were not when using seeds or sets. Inoculation of mature bulbs as a pathogenity test is another possibility. This has been tried at Lelystad but good infections using different isolates were not produced. The creation of artificial wounds influenced the test results. Inoculation of natural or sterile soil is also not promising. In sterile soil inoculation would lead to an enormous in-
crease of the fungus in the soil and hence create an abnormal pathogen population, leading to questionable results. In natural soil there will be an interference of the natural soil flora. As an alternative we have tried to develop a pathogenity test. The procedure consists of a sterilized tube ( 25 mm with a volume of 55 ml sealed with a plastic cap, schematized in Figure 12) filled with 18 ml of a growth medium. Onion seeds treated with thiram/carbendazim were germinated under sterile conditions and transferred to the tubes after germination. As soon as the growing plants have one true leaf of 3-4 cm, an agar piece with the fungus was added to the tube. After 20 days the result was observed (onion plants dead or alive). Per isolate 7-10 tubes were used. Until now the test was giving unstable results, but it seems worthwhile to further develop this test procedure as it is quick and cheap. Another possibility is to determine the pathogenity of isolates of Fusarium oxysporum f.sp. cepae with the PCR (Polymerase Chain Reaction) technique and DNA fingerprinting. The ability of Fusarium oxysporum isolates to cause basal rot is based on genetic characteristics. Individuals that have a certain behavior in common (such as pathogenicity) also have some part of their genetic material in common. If this part is known then the DNA of a certain isolate can be screened for similarity and thus for its pathogenic ability. This procedure is called DNA fingerprinting. The PCR technique is used to multiply the DNA part that needs to be detected so that small amounts of DNA amidst other material can be detected. This technique has been used widely on plant pathogens including Fusarium oxysporum. For example a testing procedure was developed by Chiocchetti et al. (1999) for detection of Fusarium oxysporum f.sp. dianthi, causing wilt in carnation. Six out of eight known races were detected on carnation cuttings. Identification of 98% of isolates of Fusarium oxysporum canariensis (palm wilt) in plant tissue was reached by Plyler et al. (1999) while the PCR technique in combination with DNA fingerprinting was used to detect Fusarium oxysporum f.sp.basilici on basil (Chiocchetti et al. 2001). AlvesSantos et al. (2002) used the PCR technique to distinguish pathogenic from non pathogenic Fusarium oxysporum on common bean (Phaseolus vulgaris) and reached a 100% identification in roots and stems of bean plants. De Haan et al. (2000) developed a multiplex PCR assay that gave good discriminating between field isolates of Fusarium oxysporum f.s. gladiolus of races 1 and 2 detected on Gladiolus corm material. This PCR assay is currently used by the Bulb Inspection Service when suspected corms are found in The Netherlands. There are no reports of this technique being applied to Fusarium oxysporum f. sp. cepae. Nevertheless, the PCR procedure has huge promise to develop detection procedures for Fusarium oxysporum f.sp. cepae in planting material as well as soils, because of its reliability and cost-effectiveness.
Research which is needed - To manage fusarium basal rot properly in the future several tools should be available: - Varieties with good tolerance level and at the same time good yield and quality potential. The search for good varieties should continue together with the breeding companies. For infected fields, these tolerant varieties would be of great help. - A reliable and cost-effective detection procedure for both soil samples and plant samples (sets). The PCR plus DNA fingerprinting technique has good promise to fill this gap as witnessed by successes with other formae speciales of Fusarium oxysporum. The further spread of the disease could then be reduced and sets could be certified as being safe in terms of infection. - As crop rotation is considered an important tool in basal rot disease management, more should be known about the population dynamics of the fungus. So far research in this direction has not been carried out, probably resulting from the lack of a reliable detection method. Development of a PCR plus DNA fingerprinting technique seems a prerequisite to these studies. Related items such as host range, survival rate in the absence of host plants and reproduction capacity on host plants, could be addressed to produce management tools on field and farm level. - Measures such as organic soil amendments have shown good promise of providing the onion industry with effective control tools. It is recommended that more detailed research is done on this direction.
REFERENCES Abawi G.S., Lorbeer J.W. 1971a. Populations of Fusarium oxysporum f.sp. cepae in Organic Soils in New York. Phytopathology 61: 1042-1048. Alabouvette C., Lemanceau P., Steinberg C. 1993. Recent Advances in the Biological Control of Fusarium Wilts. Pestic. Sci. 37: 365-373. Alves-Santos F.M., Ramos B., Asuncin Garca-Snchez M., Eslava A.P., DazMnguez J-M. 2002. A DNA-based procedure for in planta detection of Fusarium oxysporum f.sp. phaseoli. Phytopathology 92(3): 237-244. Blok W.J., Bollen G.J. 1996. Etiology of asparagus replant-bound early decline. European Journal of Plant Pathology 102: 87-98. Blok W.J., Zwankhuizen M.J., Bollen G.J. 1997. Biological Control of Fusarium oxysporum f. sp. asparagi by applying non-pathogenic isolates of F. oxysporum. Biocontrol Science and Technology 7: 527-541. Cebolla V., Martinez P.F., Dusto del A., Cases B. 1993. Control de Fusarium oxysporum f. sp. dianthi mediante solarizacio combinada con fumigantes a bajas dosis. Actas Horticultura 9: 552-557. Chiocchetti A., Sciaudoe L., Durand F., Garibaldi A., Migheli Q. 2001. PCR detection of Fusarium oxysporum f.sp. basilica on basil. Plant Dis. 85: 607-611. Haan de L.A.M., Numansen A., Robroeck E.J.A., Doorn van J. 2000. PCR detection of Fusarium oxysporum f.sp. gladioli race 1, causal agent of Gladiolus yellows disease, from infected corms. Plant Pathol. 49, 89-100. Komada H. 1975. Development of a Selective Medium for Quantitative Isolation of Fusarium oxysporum from Natural Soil. Rev. Plant Protec. Res. 8: 114-125.
Kommedahl, T., Christensen J.J., Frederiksen R.A. 1970. A Half Century of Research in Minnesota in Flax Wilt Caused by Fusarium oxysporum. Agricultural Experiment Station, Technical Bulletin 273, University of Minnesota. Mace M.E., Bell A.A., Beckman C.H. 1981. Fungal wilt disease of plants. Academic Press: 51-80. Nelson P.E. 1981. Life Cycle and Epidemiology of Fusarium oxysporum. In: Mace, M.E., A.A. Bell & C.H. Beckman (eds). Fungal Wilt Diseases of Plants. Academic Press, New York etc. pp. 51-80. Ozer N., Koycu N.D., Mirik M., Soran H., Boyraz D. 2002. Effect of some organic amendments on onion bulb rot. Phytoparasitica 30(4): 429-433. Pataky N.R. 1988. Fusarium wilt diseases of herbaceous ornamentals. Report on Plant Disease. University of Illinois extension RPD No 650, 1-9. Plyler T.R., Simone G.W., Fernandez D., Kistler H.C. 1999. Rapid detection of Fuarium oxysporum lineage containing the Canary Island date palm wilt pathogen. Phytopathology 89(5): 407-413.
FUZARYJNE GNICIE CEBULI W HOLANDII Streszczenie Fuzaryjne gnicie cebuli staje si coraz groniejsz chorob wystpujc w praktyce nawet w krajach o umiarkowanych warunkach klimatycznych. Przeprowadzone w Holandii badania wykazay, e s odmiany cebuli tolerancyjne na t chorob, aczkolwiek uzyskuje si nisze plony charakteryzujce si mniejsz przydatnoci do dugotrwaego przechowywania. Odmiany te mog stanowi jeden z elementw strategii walki z t chorob. Innymi sposobami zapobiegajcymi poraeniu plantacji cebuli przez Fusarium s: unikanie uprawy cebuli na zakaonych polach, stosowanie odpowiedniego podozmianu, zachowanie wysokiej higieny produkcyjnej w gospodarstwie, wykorzystywanie tylko zdrowego materiau (nasion i dymki), usuwanie z plantacji rolin poraonych, stosowanie nawoenia organicznego. Dla zmniejszenia strat spowodowanych przez Fusarium konieczne jest zastosowanie odpowiedniej strategii walki z t chorob. Bdzie to moliwe po opracowaniu niezawodnych i stosunkowo tanich metod i procedur oznaczania zakaenia gleby i rolin tym patogenem.

RESEARCH INSTITUTE OF VEGETABLE CROPS SKIERNIEWICE
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EFFECTS OF WATER SUPPLY METHODS AND INCUBATION DURING PRIMING ON GERMINATION OF TOMATO (LYCOPERSICON ESCULENTUM MILL.) SEEDS
Bogumia BADEK1, Bert van DUIJN2, Mieczysaw GRZESIK1 1 Research Institute of Pomology and Floriculture, ul. Pomologiczna 18, 96-100 Skierniewice, Poland e-mail: bbadek@insad.pl 2 TNO Applied Plant Sciences, P.O. Box 2215, 2301CE Leiden, The Netherlands
Summary
The aim of the study was to examine the influence and importance of water supply methods by seeds and their incubation during priming on germination, in order to elaborate the most effective priming technology. The experiment was carried out during 2002-2005 on a commercial tomato (Lycopersicon esculentum Mill.) seeds cv. Janosik. Tomato seeds were moistened up to 30-40% m.c. by means of the following methods: (i) soaking in excessive amount of water; (ii) soaking in limited amount of water; and (iii) matriconditioning. After moistening they were incubated at 20C for 1-12 days. Thereafter, the seeds were dried down to the initial moisture content and subject to germination. The germination percentage, time for the first seed to germinate (T1), time to reach 50% of germination (T50), its uniformity demonstrated by time between 10 and 90% of Gmax (T90-T10) and dynamics of germination at 15, 20 and 35C, were evaluated. All the priming methods improved germination parameters, as compared with the control. However, seed moistening in an excessive amount of water resulted in the most profitable results. While the effectiveness of the seed moistening in a limited amount of water and matriconditioning was slightly worse. The research showed that, irrespective of the method, moistening of tomato seeds up to 35% and then their incubation for 10 days were the most profitable for dynamics of germination and its uniformity. Moistening to the lower or higher moisture content and a shorter or longer time of incubation resulted in slower acceleration of seed germination and in its lower homogeneity. Our results showed that the level of the seed moisture content and time of their incubation proved to be a most important conditioning factor for germination. key words: tomato, seed priming, seed germination, incubation, water supply method
INTRODUCTION Preplant physiological seed conditioning has been used to improve seed performance and seedling emergence. The fact being important for growers as they are interested in applying of these methods in their plant production. Up till now several priming methods have been known, but more recently, matriconditioning with solid carriers of water and especially hydropriming have been attempting to use in practice (Woodstock 1988, Khan 1992, Copeland & McDonald 1995, McDonald 1999, 2000). All the mentioned methods involve water uptake by seeds, and then their incubation for initiation of metabolic processes determining germination. However, effectiveness of these priming methods depends on different factors, such us: the method of water supply to seeds, rates of water uptake, the level of the seed moisture content, temperature, availability of oxygen in the course of conditioning, time of incubation of the moistened seeds and several others. Influences of these factors of germination depend on the seed lot or species and, in spite of a lot of research in this field showing different results of priming, the literature concerning influence and importance of particular factors in this technology is still very scant. Owing to a lot of results which are difficult to be compared, the comparison of priming methods efficacy and optimization of decisive parameters are very important for production on a large scale. Recapitulation of research and comparison of efficiency of the chosen methods of conditioning of tomato seeds, in aspect of seed moistening up to 9.0-46.9% m.c. and incubation for 1 day, were described by Badek, et al. (2006). The research showed that, irrespective of the method, moistening of tomato seeds up to 30-40% m.c. and then incubation for 1 day were the most profitable for dynamics of germination and its uniformity. Still, the influence of prolonged time of incubation on germinability of these seeds which are moistened to the particular level of moisture content is not known. The aim of the present research was to establish the most effective method of priming and elaboration of the most important factors determining effectiveness of priming of tomato Janosik seeds, with the special attention paid to optimization of moisture content, incubation time and water supply methods to seeds. MATERIAL AND METHODS Experiments were conducted on commercial seeds of tomato (Lycopersicon esculentum Mill.) seeds Janosik harvested in Central Poland in 2001. Three methods of seed conditioning were used in the experiments. Untreated seeds served as a control group. Based on the earlier results concerning the dynamics of water supply and the level of their imbibition, the seeds were imbibed up to 30, 35 and 40% m.c. by means of the following methods: (i) Soaking in an excessive amount of water. Seeds were soaked in an excessive amount of water at 20C, at the ratio of seeds to water (v:v) of 1:3. The seeds were placed in glass bottles filled with distilled water for 30, 60 and 180 min. to obtain moisture contents of 30, 35 and 40%, respectively.
B. BADEK EFFECTS OF WATER SUPPLY METHODS ... 19 _____________________________________________________________________________________________________
In the course of soaking, the seeds were aerated by air bubbling through the water. After soaking, the seeds were surface dried between filter paper for 5 seconds and subsequently incubated for 1; 2; 4; 6; 8; 10 and 12 days in air-tight glass bottles at 20C. The bottles were opened every day for a few seconds for seed aeration. (ii) Soaking in limited amounts of water. Seeds were soaked in 320, 400 and 560 ml of water per kg of seeds to reach moisture contents of 30, 35 and 40%, respectively. Than the seeds were incubated for 1, 2, 4, 6, 8; 10 and 12 days in air-tight glass bottles at 20 C. The bottles were opened every day for a few seconds for seed aeration. (iii) Seeds were matriconditioned by mixing of seeds with Calflo (Celite Corporation P.O., Lompoc, USA) and water, and then incubated for 1, 2, 4, 6, 8; 10 and 12 days at 20 C in air tight glass bottles. The ratio of seeds to Calflo (w:w) was 1.0:0.4, while the ratio of seeds to water (w:w) was 1.0:0.6; 1.0:1.0 and 1.0:1.4, which enabled to obtain seed moisture contents of 30, 32.5, 35, 37.5 and 40%, respectively. The matriconditioned seeds were aerated every day for a few seconds. After the incubation, the seeds were washed for 5 seconds with distilled water to remove Calflo residues from their surface. Directly after conditioning, all the seeds were dried for 2 days, lying on filter paper at 20C and 40% air relative humidity (RH). Seeds conditioned and then dried, were subject to evaluation of their moisture content and germination properties expressed by means of germination percentage, mean germination time and dynamics of germination. The moisture content of seeds, conditioned by all the methods, was determined in two replications according to the commonly applied ISTA procedure (ISTA 2004). The seed samples were weighed, dried for 1 hour at 130C and then reweighed. The seed moisture content was expressed on a fresh weight basis. Seed germination was evaluated at temperatures of 15, 20 or 35C. Three replicates of 50 seeds each were sown in 9.0 cm-diameter Petri-dishes on cotton wool moistened with 6.0 ml of distilled water. Each seed was counted as germinated when a radical protruded through the seed coat. Germination (radical protrusion) was scored on a daily basis. Time for the first seed to germinate (T1), time to reach 50% germination (T50) and time between 10 and 90% of G max (T90-T10) were calculated by means of SeedCalculator version 3.0, a computer program for seed industry developed by Plant Research International, Wageningen, The Netherlands. The obtained results concerning germination percentage were statistically analyzed by means of variance analysis. The data of seed germination percentage were transformed according to y=arc sin(x) as outlined by Szczepaski & Rejman (1987) (a significance level of 5%). Standard errors (SE) for the results presented in table and figures were calculated by means of SeedCalculator 3.0.
RESULTS Germination of the primed seeds with the applied methods was evaluated at 15, 20 and 35C. Since the most distinct influence of seed priming on germination at the lowest temperature was observed, the presented results in tables and figures concern germination of seeds at 15C. The obtained results showed that the used three methods of conditioning of tomato Janosik seeds, as well as moisture content of seeds and time of their incubation had a crucial role in improvements germination properties demonstrated by time for the first seed to germinate (T1), time to reach 50% germination (T50) and uniformity - time between 10 and 90% of G max (T90-T10), as well as dynamics of germination (Table 1, Figs 1, 2, 3 & 4). These methods positively affected germination, although priming based on soaking in excessive amount of water was the most advantageous. Soaking in limited amounts of water and matriconditioning improved germination in a slightly lesser degree. Irrespective of the applied method, gradual moistening of seeds up to 35% m.c. resulted in the highest germination percentage. These seeds started to germinate earliest (T1) (Fig. 1) and the time to reach 50% of germination (T50) (Fig. 2) or the time between 10 and 90% of G max (T90-T10) (Fig. 3) were also shorter. Seeds having more water than 35% m.c. germinated to a lesser degree, later and less uniformly (Figs 3 & 4). Incubation time of the moistened seeds was the other important factor affecting the effectiveness of priming. Extended incubation time of seeds, moistened up to 35% m.c. from 1 to 10 days, positively and progressively affected dynamics and uniformity of germination. The longer incubation time than 10 days, irrespective of the moistening method, lowered germination percentage and dynamics of germination. Such parameters as moisture content of seeds and their time of incubation during priming exerted the highest influence on the speed of germination, while the method of priming affected it to the smallest degree (Figs 1, 2, 3 & 4). Using the mentioned parameters caused that germination was fastest when seeds were soaked in excessive amounts of water and slightly slower in seeds soaked in limited amounts of water or matriconditioned.
Table 1. Germination percentage at 15C of the tomato seeds imbibed up to 40% by soaking in excessive amount of water, soaking in limited amount of water or matriconditioning and then incubated for 1-12 days at 20C. Means of 3 replications S.E. Moisture content (%) of conditioned seeds Days of incubation None (Control) 1 2 4 6 8 1 12 1 2 4 6 8 10 12 1 2 4 6 8 10 12 9.9 92.0 1.15 Soaking in excessive water amount 96.2 1.15 96.0 0.00 96.2 2.00 94.7 1.76 96.2 1.15 96.2 1.15 94.1 0.67 96.0 0.00 96.2 3.71 96.0 0.00 96.2 1.15 95.4 0.67 95.7 1.76 96.7 0.67 94.7 0.67 94.7 0.67 97.6 1.76 89.7 2.91 92.0 0.67 92.3 2.31 88.1 1.15 Soaking in limited amount of water 92.1 1.15 93.6 1.76 93.8 2.67 92.1 1.15 94.0 0.00 94.1 1.15 94.1 1.15 94.1 1.15 94.1 1.15 94.7 0.67 94.7 0.67 92.2 2.00 94.8 1.33 95.4 0.67 94.1 1.15 91.4 2.00 92.1 0.00 85.2 2.40 91.3 0.67 92.0 1.15 83.3 1.19 Matriconditioning 92.0 0.00 92.0 0.00 94.9 1.29 94.1 1.15 94.1 1.15 93.4 0.67 96.4 2.91 96.0 0.00 95.1 1.12 95.0 1.76 96.7 2.31 92.9 1.76 96.7 1.76 96.2 1.33 93.4 1.91 94.0 0.00 96.9 1.33 91.8 1.91 90.8 1.33 92.0 0.00 88.2 1.31 30 35 40
Soaking in excessive water amount

1 days of incubation
100 100 80 60 40 20 0 0 2 4 6 8 10 12 14 0 2 4 6 8 10 12 14
100 80 60 40 20 0 0
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Soaking in limited water amount

100 100 80 60 40 20 0 0 2 4 6 8 10 12 14 0 2 4 6 8 10 12 14 100 80 60 40 20 0 0 2 4 6 8 10 12 14
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100 100 Gremination (%)
Gremination (%) 100 80 60 40 20 0
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80 60 40 20 0 0 2 4 6 8 10 12 14
Control 30% m.c. 35% m.c. 40% m.c.

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Days of germination
Days of germination
Fig. 4. Dynamics of germination at 15C of the tomato seeds, as affected by imbibition up to 3040 % m.c. and incubation for 1, 10 and 12 days, at 20 C. Means of 3 replicates. Vertical bars correspond to SE. Where no bars are shown the spread of SE is less than size of the symbol
DISCUSSION It is well known that the presowing priming of seeds may accelerate their germination and increase their uniformity or, in some cases germination percentage (Copeland & McDonald 1995, Jett et al. 1996, Bewley 1997, Warren & Bennett 1997, McDonald 1999, 2000, Nascimento & West 2000). In spite of the wide information in this field, a little attention has been paid to evaluation and importance of the factors which play a crucial role in efficiency of priming, as well as to elaboration on the most effective method of this treatment (Badek et al. 2006). The obtained results show that a great extent of efficiency of seeds
conditioning depends on proper conditions of seeds conditioning and a suitable selection of methods. They confirm our earliest findings (Badek et al. in press) that the level of the seed moisture content plays an important role in the process of ennobling seeds. They show that a very important factor is also the time of incubation of properly moistened seeds, in which metabolic processes, determining radical protrusion, are initiated. Effectiveness of priming also depends on the method of the seed moistening, although its influence seems to be lower than both factors mentioned above. The examined dependencies are in agreement with our earlier experiments in which tomato seeds were differently moistened to reach different moisture contents and then conditioned for one day or with seeds of China aster similarly moistened and conditioned for different periods, even up to over 10 days (Badek et al. in press). Research showed that for tomato Janosik, irrespective of priming methods, moistening of seeds up to 35.0% and then their incubation for 10 days at 20oC was the most profitable. It increased germination percentage from 92.0% (in the control) up to 97.6% and greatly accelerated radical protrusion, demonstrated by T1, T50 and uniformity - T90-T10. A gradual increase in moisture content, due to the moistening up to 35% progressively accelerated germination and increased its percentage, whereas increased water content over 35% delayed and lowered its radical protrusion. Similar independencies were observed in seeds moistened up to 35%, when incubation time was gradually prolonged from 1 to 10 days (gradually accelerated germination) and then over 10 days (worse germination). The same relationships, concerning moisture content, time of incubation and germination, were observed in China aster seeds moistened up to 37% and incubated for 8 days (Badek et al. in press). The obtained results suggest that the mentioned moisture content and incubation time were most optimal for initiation of repair processes in cell structures, which could be broken down by oxidative processes and initiation of metabolic processes which determine germination. Simultaneously higher moisture content and longer conditioning period of moistened seeds could also initiate ageing process, which results in decreased germination (Doijode 2001). Due to the factors, which have been explored in both tomato and China aster, belonging to Solanaceae and Asteraceae family, respectively, we assume that a similar relationship can be found in other plants. However, the value of the priming parameters (moisture content and incubation time) can be slightly modified by the structure of seed coat and morphology or composition of seeds, which depend on the plant species (Khan 1992, McDonald 1999, 2000). The obtained results show that effectiveness of priming can be slightly modified also by the method of seed moistening. For tomato Janosik, all the applied methods of priming affected positively seed germination; however, seed moistening in excessive amount of water was more beneficial than moistening in limited amounts of water or matriconditioning. It is exhibited by faster and more uniform germination. This improvement in germination is probably as the result of the faster water entering into seeds and uniform moistening of them at the same time (McDonald 1999, 2000).
Both the present research and investigation which have been made before on China aster and tomato (Badek et al. 2006) show that proper priming of seeds greatly improves their germination. Effectiveness of this treatment slightly depends on methods of water supply to seeds, but first of all, the most important factors influencing effectiveness of priming seem to be the level of moisture content to which seeds are imbibed and the time of their incubation. For tomato Janosik, moistening of seeds up to 35.0% and their incubation for 10 days at 20oC seem to be the most beneficial. Utilization of this method can be useful in practice and it is easy to apply, cheap and ecological. CONCLUSIONS The obtained results showed that the level of the seed moisture content and time of their incubation proved to be a most important conditioning factor for germination. Irrespective of the method of priming, moistening of tomato seeds cv. Janosik up to 35% and then their incubation for 10 days were the most profitable for dynamics of germination and its uniformity.
REFERENCES Badek B., Van Duijn B., Grzesik M. 2006. Effects of water supply methods and seed moisture content on germination of China aster (Callistephus chinensis) and tomato (Lycopersicon esculentum Mill.) seeds. European Journal of Agronomy 24: 45-51. Badek B., Van Duijn B., Grzesik M. (in press). Effects of water supply methods and incubation on germination of China aster (Callistephus chinensis) seeds. Seed Sci. Technol. Bewley J.D. 1997. Seed germination and dormancy. The Plant Cell 9: 1055-1066. Copeland L.O., McDonald M.B. 1995. Seed enhancements. pp.: 258-276. In: Seed Sci. Technol. New York: Chapman & Hall. Doijode S.D. 2001. Seed storage of horticultural crops. Food Products Press. An Imprint of The Haworth Press, Inc. New York. London. Oxford. ISTA 2004. International Rules for Seed Testing. Edition 2004. The International Seed Testing Association. Basserdorf, CH-Switzerland. Jett L.W., Welbaum G.E., Morse R.D. 1996. Effect of matric and osmotic priming treatments on Broccoli seed germination. J.Amer. Soc. Hort. Sci. 121: 423-429. Khan A.A. 1992. Preplant Physiological Seed Conditioning. Hort. Reviews 13: 131181. McDonald M.B. 1999. Seed deterioration: physiology, repair and assessment. Seed Sci. Technol. 27: 177-237. McDonald M.B. 2000. Seed priming. pp.: 257-286. In: Seed Technology and Its Biological Basis. (eds. M.Black & J.D.Bewley), Sheffield Academic Press, England. Nascimento W.M., West S.H. 2000. Drying during muskmelon (Cucumis melo L.) seeds priming and its effect on seed germination and deterioration. Seed Sci. Technol. 28: 211-215. Szczepaski K., Rejman S. 1987. Statystyka bada sadowniczych. (Statistical Analysis in Pomology). PWRiL, Warsaw, Poland. [in Polish]
Warren J.E., Bennett M.A. 1997. Seed hydration using the drum priming system. Hort. Sci. 32: 1220-1221. Woodstock L.W. 1988. Seed imbibition: a critical period for successful germination. J. Seed Technol. 12: 1-15.
WPYW METODY POBIERANIA WODY I INKUBACJI PODCZAS KONDYCJONOWANIA NA KIEKOWANIE NASION POMIDORA (LYCOPERSICON ESCULENTUM MILL.) Streszczenie Celem bada prowadzonych w latach 2002-2005 byo okrelenie wpywu metody pobierania wody przez nasiona pomidora odm. Janosik oraz czasu ich inkubacji podczas kondycjonowania na zdolno, dynamik i rwnomierno kiekowania, a nastpnie wybranie najbardziej efektywnej technologii kondycjonowania tych nasion. Nasiona uwilgotniono do 30-40% przez: (i) moczenie w nadmiarze wody; (ii) moczenie w ograniczonych dawkach wody i (iii) matrykondycjonowanie. Nastpnie, nasiona inkubowano przez 1-12 dni, w 20C. Po inkubacji wysuszono je do wilgotnoci pocztkowej (9,9%). Jako kondycjonowanych nasion oceniano pod wzgldem procentu liczbowego skiekowanych nasion w temperaturach: 15, 20 i 35C, dynamiki kiekowania (czas, po ktrym skiekowao pierwsze nasiono (T1) i czas po ktrym skiekowao 50% nasion (T50)), rwnomierno kiekowania (czas pomidzy skiekowaniem 10-90% nasion (T90-T10)). Wszystkie metody kondycjonowania korzystnie wpyny na kiekowanie nasion, w porwnaniu z kombinacj kontroln. Najbardziej efektywne byo moczenie ich w nadmiarze wody, natomiast moczenie w ograniczonych dawkach wody, a take matrykondycjonowanie, oddziaywao w nieco sabszym stopniu. Niezalenie od stosowanej metody, uwilgatnianie nasion do 35%, a nastpnie inkubowanie ich przez 10 dni w temperaturze 20C, najkorzystniej wpyno na dynamik i rwnomierno kiekowania nasion pomidora. Nisza lub wysza wilgotno nasion oraz krtszy lub duszy czas inkubacji poprawia kiekowanie nasion i jego rwnomierno w mniejszym stopniu.

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GENETIC VARIABILITY AND INTERRELATIONSHIPS BETWEEN TRAITS FOR AGRONOMIC PERFORMANCE AND SEEDS CULINARY QUALITY OF DRY BEAN LINES AND THEIR PARENTS
Lech BOROS, Anna WAWER Plant Breeding and Acclimatization Institute Radzikw, 05-870 Bonie, Poland
Summary The characteristics of major agronomic traits and seed culinary quality properties for two populations of breeding lines originated from crosses between fast-cooking and slow-cooking cultivars are presented in this paper. Population 1 consisted of 21 lines derived from Prosna x Nida while the second population with 67 lines derived from Isles x Narew. The average values of agronomic traits of both populations were not significantly different from that of respective parental forms. Similarly, average values of culinary quality traits of tested populations were in between parental genotypes. Significant variability for most of traits among tested bean lines was found and a few individual lines outperformed both parents of respective populations. The higher traits variability was observed for lines of second population. Presented data showed that dry bean cooking time may be shortened through selection. Some lines possessing desired combination of earliness, high yield potential, short cooking time with colored seed coats were isolated and will go into yield trials and, if they show satisfactory agricultural performance, will possibly be released as varieties; such lines would go also into recombination-cross program as parents. key words: dry bean, genetic variability, agronomic traits, water absorption, cooking time, traits associations INTRODUCTION Beans (Phaseolus ssp L.) are extremely diverse crop that is adapted to many niches, both agronomic and consumer preferences terms. Analysts estimate global production of dry beans to be 18 million metric tons annually. Dry beans account for 57 percent of the worlds food legume production, having twice the production and market value of chickpeas, the next leading food pulse. Another 6 million metric tons of snap beans are also produced annually. Nearly 80 percent of dry bean production occurs in the developing countries on small-scale farms. Poland with 16000 ha of cultivation area is one of the leading
producers of bean in European Union beside Italy, Spain, Portugal and Greece (FAOSTAT Clasic 2005). The long tradition of bean cultivation in Poland has an influence on relatively stable production for direct consumption and for processing. Beans are nutritionally rich, especially in protein and iron, along with being a good source of dietary fibre and complex carbohydrates. Given their nutritional quality and high consumption levels, beans make an important contribution to human nutrition. Regular consumption is now promoted by health organizations because it helps reduce the risks for diseases such as cancer, diabetes or coronary heart disease (Leterme 2002). Major efforts of dry bean breeders are concentrated on the increase of yield and its stability over a wide range of environments (Kelly et al. 1998, Boros 2003). However, consumer preferences and processors requirements need equal consideration to yield. Seed size, shape, color, and culinary traits related to preparation for consumption, such as water uptake and cooking time, are important parameters which influence the acceptability of cultivars by consumers. A cultivar of poor quality is likely be rejected by both consumers and processors, regardless of how agronomically superior it may be. Several studies have indicated genetic variability in dry bean for culinary traits (Castellanos et al. 1995, Shellie & Hosfield 1991, Elia et al. 1997, Boros 2003, Boros & Wawer 2000, Boros & Wawer 2005). The significant variation found for the evaluated traits indicated that they can be changed by selection. The objective of this study was to evaluate the agronomic performance and to assess the seed quality parameters existing among dry bean lines derived from different crosses in comparison to respective parental forms. MATERIALS AND METHODS Two populations of recombinant lines, twenty-one lines from cross Prosna x Nida and 67 lines originated from cross Isles x Narew together with the parental forms were chosen among different cross-combinations we dealt with in our program for further evaluation. The parental forms showed contrasting cooking time among the registered high yielding cultivars or lines from working collection (Boros & Wawer 2005). They differed in earliness and susceptibility to bean bacteriosis, with Nida and Narew cultivars being early maturing and tolerant to halo blight. Both Nida and Isles are slow-cooking cultivar with golden brown and dark red seed coats respectively. The agronomic performance of lines and parents were evaluated in two separate, 3 years field experiments seeded on May 10, May 9 and May 7 in 2001-2003 respectively at the Institute of Plant Breeding and Acclimatization in Radzikw, following standard agronomic practices and recommended procedures for fertility and weed control. The experiments were arranged in one-factor design with systematic check pattern. Experimental units consisted of four rows plot, 5 m2 in size for harvest, planting 50 viable seeds per m2. Seed yields were adjusted to 15% moisture. The determination of seed physical traits, water absorption and cooking time were done in the laboratory on seeds originated from the 2002 harvest.
L. BOROS, A. WAWER GENETIC VARIABILITY AND INTERRELATIONSHIPS ... 31 _____________________________________________________________________________________________________
After harvesting and air dried, beans were hand-cleaned to remove cull beans and foreign materials put into bags and stored in storage room at 4oC. The seed physical traits and the percentage of testa in seed are an average of 10 seeds in 3 replications. The percentage of water absorption of the entries was determined on the basis of replicated samples of 50 seeds. Seeds were soaked in distilled water for 18 hours at 25oC temperature. Bean cooking time was estimated with a 25 - seed Mattson pin-drop cooker (Jackson & Varriano-Marston 1981). The cooking time was calculated as the time from initial cooking until the time when 50% and 80% of pins penetrated seeds in the cooker. The Food and Feed 1250 apparatus was used for seed protein content determination. Ash content measurement was based on the AOAC Official Method of Analysis (1990). Data sets were evaluated by using the one way analysis of variance (ANOVA). T-test was used to evaluate the difference between genotypes for various attributes. Pearson correlation was used to evaluation the relationship between cooking time and physicochemical properties. RESULTS AND DISCUSSION Analysis of variance showed significant genotypic difference for all analyzed parameters in both populations of lines. The average values of agronomic traits of both populations were not significantly different from that of respective parental forms. Similarly, average values of culinary quality traits of tested populations were in between parental genotypes (Table 1 & 2). Significant variability of agronomic traits among tested bean lines was found and a few individual lines outperformed both parents among both populations. The higher traits variability was observed for lines of second population.
Table1. Results of preliminary evaluation of agronomic traits of parental cultivars and bean lines (three-year average) Entry Prosna Nida Lines avg Min- Max CV LSD P=0.05 Narew Isles Lines avg Min-Max CV LSD P=0.05 Vegetation days 104.8 99.1 101.2 98.8-104.6 1.5 1.5 97.3 110.3 101.1 93.7-112.7 8.7 6.8 Plant height cm Population 1 38.0 36.0 36.6 34-40 10.5 5 Population 2 41.2 40.3 39.5 30-49 14.7 8.3 Yield dt ha-1 29.9 31.4 30,0 24.3-33.7 8.1 2.4 32.2 27.1 30.1 23.2-39.5 16.9 6.3 Protein yield dt ha-1 6.3 6.5 6.2 5.1-7.2 9.7 0.6 7.3 6.2 6.8 5.2-8.9 17.6 1.4
Table 2. Seed quality traits of two populations of recombinant lines and respective parental genotypes TSW g Testa % 7.6 7.2 7.6 6.8-8.2 5.7 0.3 Ash % d.m. Protein % Water absorption % 103.9 98.0 99.9 92.5-106.1 3.3 0.5 112.6 111.7 103.9 87.1-117.4 7.2 8.3 Cooking time min 18.7 27.2 22.9 16.7-28.1 15.9 1.6 29.8 19.2 23.1 16.2-34.4 15.5 3.6
Entry
Prosna 401 Nida 383 Lines avg 404 Min-Max 333-479 CV 9.2 LSD P=0.05 10.5 Isles 522 Narew 388 Lines avg 436 Min-Max 363-557 CV 13.5 LSD P=0.05 40
Population 1 4.3 21.7 4.0 21.4 4.3 21.4 4.0-4.7 20.0-22.3 5.1 4.7 0.2 1.4 Population 2 8.1 5.0 22.3 7.3 4.4 22.9 7.3 4.4 22.5 6.0-8.6 3.8-5.0 20.7-25.8 9.6 6.2 4.9 0.9 0.2 1.2
Earliness is one of the traits of primary importance to dry bean production. Parental cultivars differed in respect of earliness. Shortening of vegetation period of lines was observed, such that the average number of days to maturity was lower than that of late parents. Early maturity exhibited by lines was more pronounced in population 2 than in population 1. Similarly, genotypic variation for seed and protein yield for population 2 was more pronounced and larger and smaller transgerssive segregant were present. Variation for seed yield observed in both populations were similar to that reported by others (Welsh et al. 1995, Wassim et al. 1990) for intraracial populations of inbred lines. Intergene pool recombinant inbred populations represented higher range of variation but recovering useful recombinations was less successful (Welsh et al. 1995, Johnson & Gepts 1998). However, due to undesirable phenotypes for other traits, for short term breeding program parents of good adaptation should be chosen besides the possibility of obtaining segregating population with high average and large variability for the trait of interest should be considered as already been reported in other studies (Abreu et al. 1999, Kelly et al. 1998). In our populations a few lines outyielded the highest yielding parent were identified. Seed quality attributes included grain weight, seed coat and ash content, protein content, water absorption, conductivity readings of soaking water and finally cooking time assessed for lines and parental forms were in the range of that reported earlier (Escribano et al. 1997, Onder & Babaoglu 2001, Boros & Wawer 2000). Wide variation among lines in both population was observed (Table 2), suggesting possible transgressive segregation. The higher variability of culinary traits, with the exception of cooking time, was observed for lines of second population.
The results of this study indicated that there were significant differences between genotypes in their abilities to absorb the water. The differences in water uptake influenced seeds hydration properties, texture, palatability, and cooking time were partially associated with structure of the epicuticular wax layer of seed coat related to the chemistry of the Asp and asp allele as it was demonstrated in the studies by Bushey et al. (2000, 2001 & 2002). Lines with opaque seed coats tended to imbibe more water, at faster rate and color testa lines were more prone to pigment leaching than those with the shiny seed coat. Our observations agreed with the finding of Bushey et al. (2000 & 2004). Cooking time exhibited a wide range from 17 to 28 min and form 16 to 34 min between lines in population 1 and population 2 respectively. In general, it is rated as medium to good cooking quality compared to dry bean cultivars cultivated in Poland (Boros & Wawer 2000, 2005). Reduction in cooking time is important from the point of view of convenience and nutritional value. Prolonged cooking of pulses has been reported to increase the percentage of leached solids, to destroy heat-labile vitamins and to decrease of protein quality. The results from the present study suggest that seed mass, thickness of seeds coat and others properties not measured here, such as composition of cotyledons may play a role in influencing cooking time. Phenotypic correlation analyses were performed to determine association among seed size, testa and ash content and culinary traits related to preparation for consumption, such as water uptake and cooking time (Table 3). Correlation coefficients among pairs of characters indicated, with few exceptions, that there were low associations among quality traits also reported by Ghaderi et al. (1984), Lu & Chang (1996), Escribano et al. (1997), Onder & Babaoglu (2001), Asensio-S.-Manzanera et al. (2005). The cooking time in both populations of lines was negatively associated with water absorption, conductivity reading and with TSW only in population 1. However, significant positive association was demonstrated between cooking time and testa and ash content as well as between testa and ash content. The relation between cooking time and water absorption (from 0.29 to 0.54) has indicated that, to some degree, the recombinant lines, especially form population 1, with higher water absorption were faster to cook. Our data, similar to the earlier reports (Castellanos et al. 1995, Shellie & Hosfield 1991, Elia & Hosfield 1995, Elia et al. 1997, Boros & Wawer 2000), also showed that the accessions with white seed coats and higher water absorption cooked comparatively faster than parental genotypes and lines that with color seed coats. The longest cooking time may be caused by tannin content in the testa. The data on cooking time were similar to those of Elia et al. (1997), who found that low tannin beans cooked faster than beans with high tannin.
Table 3. Correlation coefficients indicating relationships among culinary quality traits of population 1 (upper triangle) and population 2 (lower triangle) of dry bean lines Traits* 1. TSW (g) 2. Water absorption (%) 3. Conductivity (S cm-1g-1) 4. Cooking time 1 (min) 5. Cooking time 2 (min) 6. Testa content (%) 7. Ash (% of d.m.) 8. Protein content (%)
x
1 0.238 xx
2 3 4 5 6 7 8 xx xx xx xx 0.417 -0.444 -0.362 -0.361 -0.042 -0.083 0.322 xx 0.155 -0.536 xx -0.535 xx 0.049 -0.130 0.351 xx n.a. n.a. n.a. n.a. n.a.
-0.427 xx -0.027 0.018
-0.242xx -0.288 xx
0.984xx 0.447 xx 0.457 xx -0.233 0.447 xx 0.457 xx -0.233 0.692xx -0.054 -0.095 -
-0.092 -0.360 xx -0.186 xx 0.904xx 0.085 -0.032 0.120 0.107 0.093 0.016 -0.087 0.082 -0.159
x
0.239xx 0.244xx
0.185 xx 0.186 xx 0.408 xx -0.127
-0.111 -0.056 -0.121
and xx significant at 5% and 1% levels respectively * - traits numbers in column correspond to those in the row n.a. - not available
The intermating late maturing high yielding and fast-cooking cv. Prosna with early maturing, high yielding, golden brown seeded cv. Nida and late maturing, slow-cooking, dark red kidney cv. Isles with early maturing comparatively fast cooking cv. Narew provided recombinants with improved yield, earliness and fast-cooking. Three lines from recombinant lines population 1 (FS 5348, F 5322 and F 5404) and several lines from population 2 (FS-3124, FS 3205, FS 3171, FS 3070, FS 3165, FS 3210) were identified that have shorter or equal cooking time, that were also earlier and had slightly improved yield comparing to the maternal parent. The most of them exhibit colored seeds, which because of presence and amounts of flavonol glycosides, anthocyanins and condensed tannins in seed coats as reported by Beninger & Hosfield (2003) and Druyska (2002), may be an important source of dietary antioxidants. Presented data showed that dry bean cooking time may be shortened through selection. High productive, early maturing, color seeded and fast cooking recombinant lines were isolated. CONCLUSION Progress in a breeding program depends on the choice of parents to be involved in hybridization. The present work shows that average values of agronomic and culinary quality traits of both populations of bean line were not significantly different from that of respective parental forms. On the other hand, significant variability for most of traits among tested bean lines was found and a few individual lines outperformed both parents of respective populations.
The cooking time in both populations of lines was negatively associated with water absorption, conductivity reading and with TSW only in one population. Moreover, significant positive association was demonstrated between cooking time and testa and ash content. The relation between cooking time and water absorption indicated that recombinant lines with higher water absorption were faster to cook. The white seeded lines showed the shortest cooking time among lines in both populations. However, different market classes colored seeded lines with shorter cooking time in comparison with colored seeded parent were found. This indicates that despite of unfavourable association between cooking time and tannin content in seed it is possible to select recombinant line with no evidence of that type of association. These lines will be further evaluated and if they show satisfactory performance, will possibly be released as varieties; such lines would go also into recombination-cross program as parents.
REFERENCES Abreu A.F.B., Ramahlo M.A.P., Ferreira D.F. 1999. Selection potential for seed yield from intra- and inter-racial populations in common bean. Euphytica 108: 121-127. Asensio-S.-Manzanera M.C., Ibeas A., Fernndez S., Lpez R., Asensio C. 2005. Environmental influence on cooking time. Ann. Rep. Bean Improv. Coop. 48: 44-45. AOAC 1990. Official methods of analysis, (15th ed.) Washington, DC: Association of Official Analytic Chemists. Beninger C.W., Hosfield G.L. 2003. Antioxidant activity of extracts, condensed tannin fractions, and pure flavonoids from Phaseolus vulgaris L. seed coat color genotypes. J. Agric. Food Chem. 51(27): 7879-7883. Boros L., Wawer A. 2000. Quality characteristic of the seed of dry bean (Ph.vulgaris L.) cultivars grown in Poland. Ann. Rep. Bean Improv. Coop. 43: 93-94. Boros L. 2003. Variation in physical and chemical properties and cooking time of dry bean seeds. Veget. Crops Res. Bull. 58: 63-68. Boros L., Wawer A. 2005. Assessment of seasonal and genotypic effects on yield, quality parameters of seed and cooking time of dry edible beans (Phaseolus vulgaris L.). Veget. Crops Res. Bull. 63: 45-53. Bushey S.M., Harris L.S., Hosfield G.L. 2004. Development of an early generation test for predicting color loss of black beans. Ann. Rep. Bean Improv. Coop. 47: 139-140. Bushey S.M., Hosfield G.L., Beninger C.W. 2000. Water uptake and its relationship to pigment leaching in black beans (Phaseolus vulgaris L.). Ann. Rep. Bean Improv. Coop. 43: 104-105. Bushey S.M., Hosfield G.L., Owens S. 2002. The role of the epicuticular waxy layer in water movement across bean seed coat. Ann. Rep. Bean Improv. Coop. 45: 12-13. Bushey S.M., Owens S., Hosfield G.L.. 2001. The epicuticular waxy layer and water uptake in black beans. Ann. Rep. Bean Improv. Coop. 45: 159-160. Castellanos J.Z., Guzman-Malando H, Acosta-Gallegos J.A, Kelly J.D. 1995. Effects of hardshell character on cooking of common beans grown in semiarid highlands of Mexico. J.Sci. Food Agri. 69: 437-443. Druyska B. 2002. Polyphenolic compounds of bean seed coats (Phaseolus vyulgaris L.) and their antioxidant properties. Pol. J. Food Nutr. Sci. 11/52, 4: 35-39.
Elia F.M., Hosfield G.L. 1995. Genetic variability for cooking time of dry seeds of common bean and its relation to water absorption. Ann. Rep. Bean Improv. Coop. 38: 91-92. Elia F.M., Hosfield G.L, Uebersax M.A. 1997. Genetic analysis and interrelationships between traits for cooking time, water absorption, and protein and tannin content of Anden dry beans. J. Amer. Soc. Hort. Sci 122(4): 512-518. Escribano M.R., Santalla M., Ron A.M. de 1997. Genetic diversity in pod and seed quality traits of common bean populations from northwester Spain. Euphytica 93: 71-81. FAOSTAT Clasic 2005. http://faostat.fao.org/ Ghaderi A., Hosfield G.L., Adams M.W., Uebersax M.A. 1984. Variability in culinary quality, component interrelationships, and breeding implications in navy and pinto beans. Amer. Soc. Hort. Sci. 109(1): 85-89. Jackson G.M., Varriano-Marston E. 1981. Hard to cook phenomenon in beans: Effect of accelerated storage on water absorption and cooking time. J. Food Sci. 46: 799-803. Johnson W.C., Gepts P. 1999. Segregation for performance in recombinant inbred populations resulting from inter-gene pool crosses of common bean (Phaseolus vulgaris L.). Euphytica 106: 55-56. Kelly J.D., Kolkman J.M., Schneider K. 1998. Breeding for yield in dry bean (Phaseolus vulgaris L.). Euphytica 102: 343-356. Leterme P. 2002. Recommendation by health organizations for pulse consumption. British J. Nutr. 88, Suppl. 3: 239-242. Lu W., Chang K.C. 1996. Correlations between chemical compositions and canning quality attributes of navy bean (Phaseolus vulgaris L.). Cereal Chemistry 73(6): 758-787. Onder M., Babaoglu M. 2001. Interactions amongst grain variables in various dwarf dry bean (Phaseolus vulgaris L.) cultivars. J. Agron. Crop Sci. 187: 19-23. Shellie K.C., Hosfield G.L. 1991. Genotype x environmental effects on food quality of common bean: Resource-efficient testing procedures. J. Amer. Soc. Hort. Sci. 116(4): 732-736. Wassim N.N., Hosfield G.L., Uerbersax M.A. 1990. Inheritance of physico-chemical seed characters related to culinary quality in dry bean. J. Amer. Soc. Hort. Sci. 115: 492-499. Welsh W., Bushuk W., Roca W., Singh S.P. 1995. Characterization of agronomic traits and markers of recombinant inbred lines from intra- and interracial populations of Phaseolus vulgaris L. Theor. Appl. Genet. 91: 169-177.
ZRNICOWANIE GENETYCZNE ORAZ WSPZALENOCI CECH ROLNICZYCH I PARAMETRW KULINARNYCH NASION FORM RODZICIELSKICH I LINII REKOMBINACYJNYCH FASOLI KAROWEJ NA SUCHE NASIONA Streszczenie Porwnano dwie populacjie linii rekombinacyjnych fasoli oraz formy rodzicielskie pod wzgldem cech rolniczych i parametrw technologicznych nasion. Populacja 1 to linie wyprowadzone z krzywki Prosna x Nida, n=21, oraz populacja 2 to linie wyprowadzone z krzywki Isles x Narew, n=67. Obie populacje charakteryzoway si wartociami rednimi cech rolniczych nie rnicymi si istotnie od odpowiednich
wartoci form rodzicielskich. Podobnie, rednie wartoci parametrw jakociowych nasion badanych populacji linii fasoli byy w przedziale wartoci form rodzicielskich. W szeregu przypadkach badane linie przewyszay pod wzgldem okrelonej cechy lepszego rodzica. Wystpiy rnice w zalenociach pomidzy cechami badanych populacji linii fasoli. Stwierdzono znaczny zakres zmiennoci badanych cech, pozwalajcy na wybr linii o korzystnych cechach. Wysz zmienno badanych cech stwierdzono w populacji 2 linii fasoli. Wytypowano linie z obu populacji, czce wysoki poziom plonowania, wczenie dojrzewajce, o krtszym czasie gotowania i kolorowej okrywie nasiennej.

VEGETABLES SEEDS NUTRITIONAL ASPECTS IN RESPONSE TO GERMINATION TIME

Honorata DANILCENKO 1, Zivile TARASEVICIENE 1, Elvyra JARIENE 1, Marek GAJEWSKI 2, Pawe SZYMCZAK 2, Anna SEROCZYSKA 2 1 Lithuanian University of Agriculture, Kaunas, Lithuania Studentu g. 11, Kaunas, Lithuania, e-mail: hd@lzuu.lt 2 Warsaw Agricultural University Nowoursynowska 166, 02-787 Warszawa, Poland
Summary In this work the chemical composition, mineral constituents, amino acids profile, fatty acids composition of radish and alfalfa seeds were studied in order to evaluate their nutrition value in response to germination time. Radish and alfalfa seeds were germinated for 1, 3 and 5 days in the dark, at the temperature of 25C. Seeds germination caused the decrease in dry matter content. The moisture content of raw radish seeds was 6.45%, alfalfa seeds 8.20%, while in five days germinated radish seeds 85.34% and in alfalfa sprouts 90.36%. During germination crude proteins content showed the tendency to increase, while crude fats to decrease. After germination, content of eight amino acids decreases in radish sprouts, as well as in alfalfa sprouts. After germination of radish seeds threonine, serine, glutamic acid, glycine, alanine, leucine, lysine and arginine contents decreased while in alfalfa tyrosine, threonine, serine, glutamic acid, glycine, leucine, lysine and arginine decreased. Fatty acids composition during seeds germination changed slightly. The biggest decreasing tendency during 5 germination days of alfalfa seeds was observed in case of linolenic fatty acid (C 18:3) by 5.15%, and oleic (C 18:1) by 0.51%. In radish seeds, amount of linolenic fatty acid (C 18:3) increased by 1.56%, and of linoleic acids (C 18:2) by 1.67%. Total carotenoids and -carotene content in radish sprouts after 3 days of germination was very high (2.31 and 0.61 mg100g-1, respectively) and several times higher than in alfalfa sprouts. key words: biochemical changes, germination, nutrition value, seeds, sprouts INTRODUCTION In recent years human health has assumed a very important status. Some foods have now assumed the status of functional foods, which should be capable of providing additional physiological benefit, such as preventing or delaying onset of chronic diseases, as well as meeting basic nutritional requirements (Charanjit et al. 2001). Sprouting of seeds is one of the processing methods to
increase the nutritive value and the health qualities of foods in a natural way (Bau et al. 1997). This method has been known for a very long time, mainly in the Eastern countries (China and Japan). Germination process is simple and inexpensive, and different seeds have been germinated for human consumption, such as legumes (soybean, lentils, beans, chickpeas, peas), cereals (wheat, rye, barley, oats), and recently, seeds of some other vegetables (alfalfa, radish, mustards, cabbage) (Frias et al. 2005). Alfalfa and radish seeds are the most popular vegetable seeds used for germination, because of the good sensory features as well as they are rich source in antioxidants. During germination process seed storage compounds extensively break down and new cell components arrive (Kuo et al. 2004). In comparison to seeds and seed products, sprouts have a greater nutritive value due to a better quality of proteins, a more favorable distribution of amino acids, a higher content of poly-unsaturated fatty acids, an increased bio-availability of essential minerals and trace elements, and also higher content of vitamins (Meier-Ploeger 1990). Changes in nutrients occurring during seed germination depend on the species and on sprouting conditions (i.e. time, temperature, light cycle) (Sierra & Vidal-Valverde 1999). The aim of this work was to study the effect of different germination time on the nutritional quality changes in germinating seeds of radish and alfalfa. Seeds of these species are commonly used for germination and sold on a market as vegetable sprouts. MATERIALS AND METHODS Alfalfa (Medicago sativa cv. Europa) and radish (Raphanus sativus cv. Cherry Belle) seeds were purchased in Lithuania and used for germination experiments. Germination process was carried out in special sprouters Bionatura. Seeds were washed and soaked in tap water for 12 hours. Imbibed seeds were sprouted for 1, 3 and 5 days in the dark at 25C. Every 24 hours seeds were irrigated with water. Germinated seeds were dried in an electric air drought oven at 65C for 24 hours and milled. Dry matter content of the dry and sprouted seeds was determined gravimetrically by drying to constant weight at 105C. Crude proteins, crude fats, ash, crude fibre was determined in raw and germinated 1, 3 and 5 days seeds (Nauman & Bassler 1993). Amino acids were determined using a Mikrotechna AAA 339 automatic amino acids analyzer according to the normative act compendium (Anonymous 2003). Hydrolysis of the sample was performed in the presence of 6 M HCl at 105C for 24 h. Fatty acids were extracted by method described by Folch. Extracted composite of fatty acids and methyl ester analysed using gas chromatography (GC-2010 Shimadzu), with hydrogen flame detector (Folch 1957, Christopherson & Glass 1969). -carotene and total carotenoids content was determined by the spectrophotometer UV-1201V Shimadzu, using the wavelength 450 nm (Lichtenthaler & Wellburn 1983, Rutkowska 1981). Results are expressed as the mean values of four replicates (two plates of germinated alfalfa and radish seeds). Data were statistically analyzed using
H. DANILCENKO et al. VEGETABLES SEEDS ... 41 _____________________________________________________________________________________________________
analysis of variance (ANOVA) and least significant difference, using SigmaStat 2.03. Significant differences were determined at the P<0.05. RESULTS AND DISCUSSION During alfalfa and radish seeds germination there was detected a noticeable decrease in dry matter content. After 5 germination days dry matter content in germinated alfalfa seeds was equal to 9.93% and in radish seeds 17.88% (Fig. 1). The most intensive decrease in dry matter content was detected after first 24 germination hours in alfalfa and in radish sprouts. At different germination stages there was observed higher amount of dry matter in radish seeds, compared with alfalfa seeds (Fig. 1). Significant decrease in dry matter content is related to water absorption during seeds soaking and degradation or oxidation of starch and sugars during respiration, to provide energy for the increased metabolic functions in the sprouting seed. The respiration process produces carbon dioxide, which escapes from the seed (Mubarak 2005). In can be concluded that alfalfa seeds absorbed more water than radish seeds.
100 90 80 70 60 % 50 40 30 20 10 0 Raw 1 3 5 Radish Alfalfa
Germination time (days)
Fig. 1. Dry matter content during alfalfa and radish seeds germination. Vertical lines mean standard deviation for each combination
Chemical composition of raw and germinated alfalfa and radish seeds are presented in Fig. 2 & 3. Germination of alfalfa and radish seeds resulted in increase of crude protein content compared with the raw seeds. This increase was mainly due to the use of seed components during the germination process. Germination resulted in decreasing of fats and nitrogen free extractives content (Fig. 2 & 3). These decrease can be attributed to their use as an energy source for faster germination. These results are in agreement with those reported by Mubarak (2005) for germinated mung-bean seeds.
100%
80% Ash 60% Nitrogen free extractives Crude fibre 40% Crude fat Crude protein 20%
0% Raw 1 3 5 Germination time (days)
Fig. 2. Chemical composition of raw and germinated alfalfa seeds
100%
80% Ash 60% Nitrogen free extractives Crude fibre 40% Crude fat Crude protein 20%
0%
Raw
Germination time (days)
Fig. 3. Chemical composition of raw and germinated radish seeds
Ash content during germination process showed only unsignificant changes in alfalfa and radish seeds. These results are in agreement with those reported by Oloyo (2004) for germinated pigeon pea and in opposition to Mubarak (2005) for germinated mung-bean seeds, and Colmenares & Bressani (1990) for germinated amaranthus seeds. The reason of this phenomenon may be taxonomical characteristics of these species. In raw alfalfa and radish seeds the major protein amino acids found were glutamic acid, aspartic acid and arginine (Table 1 & 2). Of all amino acids detected in alfalfa seeds the smallest was the amount of methionine, while in rad-
ish seeds of histidine, but the amount of amino acids fluctuated during germination. However, a scale of these changes depended on amino acid. The biggest amount of methionine in alfalfa seeds was found after third germination day and in radish seeds after one day. Amino acid tyrosine content in alfalfa seeds did not change during germination, while that of threonine increased after five germination days, valine after three germination days, compared with raw seeds. Tyrosine, threonine and valine contents did not change during five germination days in radish seeds compared to raw seeds (Table 2). Germination of alfalfa seeds resulted in significant (P<0.05) increasing of amino acids leucine, isoleucine and lysine content after three germination days, compared with the raw seeds (Table 1). The biggest content of amino acids leucine and isoleucine in germinating radish seeds was observed after one germination day, while lysine in raw seeds (Table 2). Among non-essential amino acids presented in alfalfa seeds, arginine content decreased after five germination days by 0.89%, glycine by 0.57%, serine by 0.40%, and glutamic acid by 1.61%. Aspartic acid increased by 1.72% after five days of alfalfa seeds germination (Table 1). These changes were estimated as significant.
Table 1. Effect of germination time on amino acids composition in alfalfa seeds Amino acids composition (g100 g -1 d.w.) Germination time (days) Raw seeds 1 3 5 0.81a 0.77a 0.84a 0.73a 0.86a 1.02a 1.10a 0.93a 1.20a 0.86ab 1.02ab 0.77b 0.28a 0.32ab 0.47b 0.39ab 1.88ab 1.89ab 2.01a 1.45b 0.87ab 0.91ab 1.01a 0.78b 1.03ab 1.15ab 1.16a 0.82b 0.99a 1.02ab 1.17b 1.08ab 2.95ab 2.66a 2.76ab 4.67b 4.58a 4.08bc 3.98c 2.97d 1.29a 0.96ab 1.24ab 0.89b 1.30a 0.12ab 1.22ab 0.73b 1.08ab 1.02a 1.75b 1.25ab 0.71a 0.46b 0.64ab 0.69ab 3.02a 2.76ab 2.75ab 2.13b 2.16 : 1 2.08 : 1 1.99 : 1 1.86 : 1
Amino acid Tyrosine Phenylalanine Threonine Methionine Leucine Isoleucine Lysine Valine Aspartic acid Glutamic acid Serine Glycine Alanine Histidine Arginine Leucine/isoleucine ratio
Note: means in the same row marked with different letters differ significantly (P=0.05)
After five days of radish seeds germination, content of non-essential amino acid glycine decreased by 0.42%, serine by 0.19%, and glutamic acid by 1.17%. The content of aspartic acid showed small tendency to increasing (by 0.04%), compared to raw seeds (Table 2).
Table 2. Effect of germination time on amino acids composition in radish seeds Amino acid Tyrosine Phenylalanine Threonine Methionine Leucine Isoleucine Lysine Valine Aspartic acid Glutamic acid Serine Glycine Alanine Histidine Arginine Leucine/isoleucine ratio Note: see Table 1 Amino acids composition (g100 g -1 d.w.) Germination time (days) Raw seeds 1 3 5 0.66ab 0.68a 0.66ab 0.64b 0.93ab 1.22a 0.93ab 0.82b 0.98a 0.95a 1.00a 0.79a 0.68a 0.95b 0.58ac 0.51c 1.74ab 1.81a 1.71ab 1.37b 0.85ac 1.09b 0.92a 0.76c 1.12a 0.87ab 1.03ab 0.77b 1.08ab 1.25a 1.17ab 1.07b 1.80a 2.52b 1.87ab 1.84ab 4.66ab 6.41a9 3.98ab 3.49b 0.85ab 1.02a 0.87ab 0.66b 1.13ab 1.30a 0.96ab 0.71b 1.09a 1.38ab 1.57b 1.44ab 0.65ab 0.61a 0.67b 0.62ab 1.82ab 2.46a 1.51ab 1.03b 2.05 :1 1.66 : 1 1.86 : 1 1.80 :1
During the germination of alfalfa seeds leucine / isoleucine ratio decreased. Vidyanath-Jha et al. (1991) found that the high leucine / isoleucine ratio was responsible for low biological value of aquatic crops. Excess leucine content in food interferes with the utilization of isoleucine and lysine (Mbitki et al. 2000). When taken as a major source of protein in the diet, any protein with a leucine / lysine ratio less than 4.6 is considered nutritionally safe, because higher levels of leucine in a diet may induce pellagra and the leucine / lysine ratio is used as an indicator of the pellagragenic character of a food protein (Harmut-Hoene et al. 1987). In germinated alfalfa seeds this ratio ranged from 1.82 to 1.65, and in radish seeds from 1.55 to 2.08, depended on germination period (Table 1 & 2). Fatty acids composition during alfalfa germination changed slightly (Table 3). The biggest decrease during seeds germination was observed in case of linolenic acid (C 18:3) 5.15% after five seeds germination days, and oleic (C 18:1) 0.70% after three germination days (Table 3). In case of radish seeds, linolenic acids content (C 18:3) increased during germination by 1.56%, and linoleic acid (C 18:2) by 1.67%. Palmitic acid and oleic acid (C 18:1) content increased after one germination day compared with raw seeds and then decreased after five germination days (Table 4). Total carotenoids and -carotene content was high in radish seeds (2.31 and 0.61 mg100 g-1, respectively), and higher then in alfalfa seeds (Fig. 4). Carotenoids content showed the tendency to decrease during germination process.
This work is a part of a bigger research program, so experiments will be continued with more species included, and other nutritional aspects will be investigated.
Table 3. Effect of germination time on fatty acids composition in alfalfa seeds Fatty acids composition (% of total fats content) Germination time (days) Raw seeds 1 3 5 0.16a 0.17ab 0.16ab 0.37b 0.08a 0.00a 0.00a 0.57a 10.68a 11.50a 10.68a 11.81a 0.07a 0.00a 0.11a 0.00a 0.06a 0.00a 0.00a 0.47a 0.00a 0.00a 0.00a 0.41a 2.29a 2.51ab 2.51ab 2.76b 12.01a 12.53ab 12.71b 12.52ab 42.09a 42.04a 42.02a 43.17a 28.59a 26.81ab 27.24ab 23.44b 0.68a 0.67a 0.76a 0.75a 0.20ab 0.19a 0.21ab 0.25b 0.00a 0.37a 0.23a 0.00a 0.12a 0.12b 0.14b 0.00c 0.00a 0.00a 0.00a 0.94a 0.60a 0.56a 0.83a 0.89a 0.00a 0.18a 0.11a 0.00a 0.16a 0.55a 0.49a 0.00a 0.94a 1.27a 1.32a 0.00a 0.11a 0.12ab 0.22ab 0.37b 1.18a 0.41b 0.26c 0.00d 0.00a 0.00a 0.00a 1.28a
Fatty acid C 14:0 C 15:0 C 16:0 C 16:1 C 17:0 C 17:1 C 18:0 C 18:1 C 18:2 C 18:3 C 20:0 C 20:1 C 21:0 C 20:4 C 20:5 C 22:0 C 22:1 C 23:0 C 22:5 C 24:0 C 24:1 C 22:6 Note: see Table 1
Table 4. Effect of germination time on fatty acids composition in radish seeds
Fatty acid C 14:0 C 15:0 C 16:0 C 16:1 C 17:0 C 17:1 C 18:0 C 18:1 C 18:2 C 18:3 C 20:0 C 20:1 C 21:0 C 20:4 C 20:5 C 22:0 C 22:1 C 23:0 C 22:5 C 24:0 C 24:1
Fatty acids composition (% of total fats content) Germination time (days) Raw 1 3 5 0.06a 0.07a 0.00a 0.00a 0.00a 0.00a 0.00a 0.00a 5.23a 5.63b 5.40ab 5.43ab 0.23a 0.19ab 0.16ab 0.40b 0.00a 0.10a 0.00a 0.00a 0.00a 0.00a 0.00a 0.00a 1.34a 1.32ab 1.29ab 1.20b 26.83ab 27.85a 26.67ab 25.62b 10.48a 10.69ab 11.44ab 12.15b 6.99a 7.34ab 7.68ab 8.55b 0.86a 0.80b 0.81b 0.76c 10.24a 9.85b 9.97b 9.15c 0.27ab 0.25a 0.28ab 0.28b 0.00a 0.00a 0.00a 0.28a 0.06a 0.07a 0.00a 0.00a 0.81ab 0.72a 0.75ab 0.92b 33.65a 32.45a 32.72a 32.30a 0.23a 0.27a 0.28a 0.00a 0.00a 0.00a 0.00a 0.00a 0.79ab 0.69a 0.74ab 0.81b 1.92a 1.71b 1.83ab 1.91ab
Note: see Table 1
3 2.5 2 Radish 1.5 1 0.5 0 3 days 5 days 3 days 5 days total carotenoids beta-carotene Alfalfa
Fig. 4. Total carotenoids and -carotene content in alfalfa and radish seeds germinated for 3 and 5 days (mg100 g-1). Vertical lines show standard deviation for each combination
CONCLUSIONS 1. Germination process affects the chemical composition and nutritional quality of alfalfa and radish seeds and may slightly improve their nutritional quality. 2. Germination causes changes in content of a few amino acids and fatty acids. This lead to slight changes in overall amino acids and fatty acids profile in investigated seeds. 3. Prolonged germination time causes decreasing of total carotenoids content, as well as -carotene content both in radish and alfalfa seeds.
Acknowledgements: This study was financially supported by Lithuanian State Science and Studies Foundation, Grant Nr. C- 22/2005. REFERENCES Anonymous 2003. Aminorgi ir olakvindokso kiekio paaruose nustatymo techninis reglamentas. Paar tyrimo metodai/Normatyvini akt rinkinys. 66-77. [in Lithuanian] Bau H.M., Villaume C., Nicolas J.P., Mejean L. 1997. Effect of germination on chemical composition, biochemical constituents and antinutritional factors of soya bean (Glycine max) seeds. J. Sci. Food Agric. 73: 1-9. Charanjit K., Haris C. Kapoor. 2001. Antioxidants in fruits and vegetables the millenniums health. Intern. J. Food Sci. Technol. 36: 703-725. Christopherson S.W., Glass R.L. 1969. Preparation of milk fat methylesters by alcoholysis in an essentially nonalcoholic solution. J. Dairy Sci. 52: 1289-1290. Colmenares de Ruiz A.S., Bressani R. 1990. Effect of germination on the chemical composition and nutritive value of amaranthus grain. Cereal Chem. 67: 519-522. Folch J., Less M., Sloanc-Stanley G.H. 1957. A simple method for isolation and purification of total lipids from animal tissues. J. Biol. Chem. 226: 497-509. Frias J., Martha L. Miranda, Doblado R., Vidal-Valverde C. 2005. Effect of germination and fermentation on the antioxidant vitamin content and antioxidant capacity of Lupinus albus L. var. Multolupa. Food Chem. 92: 211-220. Harmut-Hoene A.E., Bognar A.E., Kornemann U., Diehl J.F. 1987. The influence of germination on the nutritional value of wheat, mung bean and chickpeas. Z. Lebensm. Unters. Forsch. 185: 386-393. Kuo Yu-Haey, Rozan P., Lambein F., Frias J., Vidal-Valverde C. 2004. Effect of different germination conditions on the contents of free protein and non-protein amino acids of commercial legumes. Food Chem.. 86: 537-545. Lichtenthaler H.K., Wellburn A.R. 1983. Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem. Soc. Transact. 603: 591-592. Mbitki-Mwikya S., Van Camp J., Yiru Y., Huyghebaert a. 2000. Nutrient and antinutrient changes in finger millet (Eleucine coracan) during sprouting. Lebens. Wiss. Technol. 33: 9-14. Meier-Ploeger A. 1990. Nutritional value of seeds sprouts. Ern/Nutr. 14: 317-323. Mubarak A.E. 2005. Nutritional composition and antinutritional factors of mung bean seeds (Phaseolus aureus) as affected by some home traditional processes. Food Chem. 89: 489-495.
Nauman C., Bassler R. 1993. Die chemische Untersuchung von Futtermitteln, Methodenbuch Band III, VDLUFA, Verlag Darmstadt. [in German] Oloyo R.A. 2004. Chemical and nutritional quality changes during germinating seeds of Cajanus cajan L. Food Chem. 85: 497-502. Rutkowska U. 1981. Witamina i prowitamina A. In: Wybrane metody analizy skadnikw zywnoci i wartosci odzywczej. PZWL Warszawa: 302-331. [in Polish] Sierra I., Vidal-Valverde C. 1999. Kinetics of free and glycosylated B6 vitamers, thiamine and riboflavin during germination of pea seeds. J. Sci. Food Agric. 79: 307-310. Vidyanath-Jha, Barat G.K., Jha U.N. 1991. Nutritional evaluation of Euryale ferox Salisb. (Makhana). J. Food Sci. Technol. 28 (5): 326-328.
NASIONA WARZYW - ASPEKTY YWIENIOWE W ZALENOCI OD DUGOCI OKRESU KIEKOWANIA Streszczenie Badano skad chemiczny, skadniki mineralne, skad aminokwasw i kwasw tuszczowych w nasionach rzodkiewki i lucerny, w celu okrelenia zmian wartoci odywczej w zalenoci od dugoci okresu podkiekowywania. Nasiona podkiekowywano przez 1, 3 i 5 dni w ciemnoci, w temperaturze 25C. Podkiekowywanie powodowao zmniejszenie suchej masy nasion. Zawarto wody w wieych nasionach rzodkiewki wynosia 6,45%, a w nasionach lucerny 8,20%, natomiast po 5 dniach podkiekowywania odpowiednio 85,34% i 90,36%. Podczas kiekowania zawarto protein wykazywaa tendencj wzrostow, natomiast tuszczw malejc. Zawarto omiu aminokwasw zmniejszya si w kiekach rzodkiewki, podobnie jak w kiekach lucerny. W wyniku kiekowania nasion rzodkiewki zawarto treoniny, seryny, kwasu glutaminowego, glicyny, alaniny, leucyny, lizyny i argininy zmniejszya si, natomiast w nasionach lucerny dotyczyo to tyrozyny, treoniny, seryny, kwasu glutaminowego, glicyny, leucyny, lizyny i argininy. Skad kwasw tuszczowych zmienia si niewiele podczas kiekowania. Najwikszy wzrost obserwowano dla kwasu linolenowego (C 18:3) o 5,15% i kwasu oleinowego (C 18:1) o 0,51% po 5 dniach podkiekowywania. W nasionach rzodkiewki zawarto kwasu linolenowego (C 18:3) wzrastaa podczas podkiekowywania o 1,56 %, a zawarto kwasu linolowego (C 18:2) o 1,67%. Zawarto karotenoidw ogem oraz -karotenu bya wysoka w kiekach rzodkiewki po 3 dniach podkiekowywania (odpowiednio 2,31 i 0,61 mg100 g-1) i kilkakrotnie wiksza ni w kiekach lucerny.

THERMAL PROCESSING EFFECTS ON ANTIOXIDANT CONSTITUENTS AND PROPERTIES OF TOMATOES

Janusz CZAPSKI, Justyna SZWEJDA Research Institute of Vegetable Crops Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland
Summary The effects of cold and hot brake processing of tomatoes, heating of pulp, freeze drying and baking of raw tomato slices on antioxidant properties and antioxidant constituents were determined. Thermal processing significantly decreased ascorbic acid content and markedly contributed to the increase of lycopene and total soluble phenolics content. Hot break processing considerably diminished antioxidant activity (AOA) (measured by coupled oxidation of carotene and linoleic acid) of tomato pulp by 63% and significantly increased antioxidant power (EC50)-1 (measured by scavenging of stable 2,2-diphenyl-1picrylhydrazyl (DPPH) free radical) by 47% in comparison with pulp from the cold break processing. High negative correlation between phenolics content and AOA (r = -0.851) and between total flavonoids content and AOA (r = -0.74) was found. High positive correlation between phenolics content and antioxidant power (EC50)-1 (r = 0.816) and between total flavonoids content and (EC50)-1 (r = 0.935) was also found. Freeze drying and baking of tomato slices contributed to the reduction of content of all determined components and values of antioxidant parameters in comparison with raw tomato slices. key words: tomatoes, cold and hot break processing, freeze drying, baking, antioxidant constituents and properties INTRODUCTION Preservation methods are generally believed to be responsible for a depletion of naturally occuring antioxidants in food. Processed fruits and vegetables are expected to have a lower health protecting capacity than fresh ones. This is due to the fact that most of compounds are thermal unstable. A large amount of literature deals with ascorbic acid chemical oxidation and/or thermal degradation as a consequence of blanching, cooking, pasteurization, sterilization, dehydration and freezing (Murcia et al. 2000, Zanoni et al.1999). Operation such as peeling, cutting and slicing are expected to induce a rapid enzymatic depletion of several natural antioxidants as ascorbic acid and polyphenols (Mc Carthy & Mattheus 1994).
From the other side it was reported that food processing as cooking and grinding of tomatoes might improve lycopene bioavailability by breaking down cell walls. Thermal processing disrupts the cell membranes and cell walls and releases lycopene from the insoluble portion of the tomatoes, which increases the pool of bioaccessible lycopene and improves its absorption (Gartner et al. 1997, Shi & Le Maguer 2000). In humans, lycopene bioavailability was greater from heat processed tomatoes compared to fresh ones (Stahl & Sies 1992). Heat induces isomerization of the all-trans to cis forms of lycopene and bioavailability of cis isomers in food is higher than that of all-trans isomers, which are the main configuration of lycopene of fresh tomato fruits (Shi & Le Maguer 2000). The conjugated double bound system of lycopene confers strong antioxidant activity including the ability to quench singlet oxygen and peroxyl radicals. The quenching constant of lycopene was found to be more than twice that of carotene and 10 times more than that of -tocopherol, thus making its presence in the diet important (Shi & Le Maguer 2000). According to Toor et al.(2005) ascorbic acid and phenolic compounds are the main contributors to the hydrophilic antioxidant activity of tomatoes. Declines in both total phenolics and ascorbic acid contents during processing were probably responsible for the decrease in antioxidant activity in the semi-dried tomatoes (Toor & Savage 2006). Despite a decrease in ascorbic acid an increase in the antioxidant activity has been reported in the tomato products processed at higher temperatures (Dewanto et al. 2002, Giovanelli et al. 2002, Gahler et al. 2003). Heating can induce the formation of compounds such as melanoidins in the Maillard reaction and these compounds can have some antioxidant or prooxidant effects (Nicoli et al. 1999). Changes in the antioxidant activity of tomato products are complex and depend on specific compounds present. Loss in antioxidant activity associated with decrease in lycopene and ascorbic acid contents during processing may be accompanied by increase in antioxidant activity of other components particularly phenolic and flavonoid compounds. Therefore studies characterizing changes in antioxidant constituents and antioxidant activity during thermal processing are important to understand the role that fresh and processed tomatoes may play in the human diet. MATERIALS AND METHODS Tomato fruits of Polned cultivar were obtained from a commercial grower. For cold break and hot break preparation of tomato pulp, fully ripe but not spoilt fruits were used. Tomatoes after washing were drained and sorted. Then they were cut into halves to facilitate crushing and to detect any possible disease and decay inside. A manual juice extractor was used to separate the pulp from seeds and the skin and then pulp was de-aerated under vacuum. Hot break preparation of the pulp involved processing of tomato halves at temperature approximately 90C. After separation of seeds and skin, the pulp was cooled to room temperature. Eight kg of tomato fruits for cold break preparation and three kg for hot break were divided into four replicates and tomatoes from each replicate was
J. CZAPSKI, J. SZWEJDA THERMAL PROCESSING EFFECTS ... 51 _____________________________________________________________________________________________________
processed separately. Cold break tomato pulp was subjected to four different treatments: raw, heated at 90C for 2 min, 10 min and 20 min. The raw treatment consisted of tomato pulp not thermally treated. Samples of tomato pulps (about 200 g) from each of four replicates of each treatment (including hot break preparation) were poured into polyethylene bags, sealed tightly and stored at -25C until analysis. To prepare freeze dried and baked tomato slices, raw tomato fruits after washing and draining were peeled and cut on slices 4-5 mm thick. Randomly chosen slices were freeze dried using Alpha 1 2/LD Freeze Dry system (Martin Chris GmbH) for 46 h until ice condenser temperature reached about -52C (0.04 mbar vacuum). To bake the tomatoes, randomly chosen slices were baked in a pre-heated oven at two oven temperatures: 200C for 1 h or 250C for 40 min. The raw tomato slices prepared without any thermal treatment were used as a control. Each treatment consisted of 4 replicates. All samples of each treatment were weighed and then they were packaged in polyethylene bags, sealed tightly and stored at -25C until use. Phenolic and flavonoid compounds were extracted from tomato pulp and slices using 80% (v/v) acetone. Fifty grams of pulp or raw slices of tomatoes or 3 g of freeze dried or 12 g of baked slices were homogenised with 100 ml of 80% cold acetone using Ultra-Turrax T-25 blender for 2 min (pulp) or 3 min (slices). The homogenate was filtered through Whatman No.2 filter paper on a Bhner funnel under vacuum. The residue was washed using small volume of 80% acetone and allowed it to dry for 30 min in the dark at room temperature. The dry powder was weighed, ground with mortar and pestle and then it was stored in tightly stoppered polypropylene vial at 0C in the dark until use for lycopene extraction. The acetone from filtrate was evaporated by vacuum rotary evaporator at 45C. The tomato extract was centrifuged at 12000g for 10 min and supernatant was then transferred to 100 ml volumetric flask and filled to the mark with water. Such prepared tomato extract was analysed for the content of total soluble phenolics, total flavonoids and antioxidant properties. Total soluble phenolics content was determined colorimetrically using the Folin - Ciocalteau reagent (Ragazzi & Veronese 1973) an using catechin as a standard. All values were expressed as miligrams catechin equivalents per kilogram fresh weight (mgkg-1). Total flavonoids content was measured using colorimetric method described previously by Zhishen et al. (1999) and Eberhardt et al. (2000) with small modifications. Tomato extract (0.5 ml) was mixed with 1.25 ml of distilled water in a test tube. At zero time 0.075 ml of 5% (w/v) sodium nitrite was added. After 6 min 0.15 ml of 10% (w/v) AlCl3 solution was added and allowed to stand for 5 min before 0.5 ml of 1 M NaOH was added. The mixture was brought to 2.5 ml with distilled water. The absorbance was measured at 510 nm against blank using Semco S/EC spectrophotometer. The amount of flavonoids was determined from the standard curve of catechin and expressed as miligrams catechin equivalent per kilogram fresh weight (mgkg-1). Lycopene was extracted from stored dry powder. One gram of powder was blended for 2 min with 25 ml mixture of hexane acetone (4:1 v/v) using Ultra-
Turrax T-25 blender. The homogenate was centrifuged at 2000g and organic fraction was collected. The residue was further extracted two times with 10 ml of hexane acetone mixture until it was devoid of red pigment indicating that lycopene had been completly extracted. All hexane acetone fractions were combined and washed several times with water until acetone was completely removed. Lycopene was separated from -carotene by column chromatography on aluminium oxide, using method described previously (Czapski & Saniewski 1985). The eluted lycopene fraction was evaporated to dryness in the dark at room temperature under stream of nitrogen and re-dissolved in appropriate volume of hexane for spectral measurement at 472 nm on a Semco S/EC spectrophotometer. Quantity of lycopene was calculated from published extinction coefficient (E1%1cm) being 3450 (Davies 1965) and the results were expressed as miligrams per kilogram fresh weight (mgkg-1). Ascorbic acid content was determined using 2,6-dichloroindophenol titrimetric method as described in Official Methods of Analysis (AOAC 1984). Ascorbic acid content was calculated from the standard curve and the results were expressed as miligrams per kilogram fresh weight (mgkg-1 ). Two different methods were selected for evaluation of the antioxidant activity: -carotene bleaching method based on coupled oxidation of -carotene and linoleic acid (Emmons et al. 1999), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay (Brand-Williams et al. 1995, SanchezMoreno et al. 1998, 1999). In -carotene bleaching method, aliquots (3 mL) of the -carotene and linoleic acid emulsion were mixed with tomato extract (0.1 mL) and incubated in a water bath at 50C for 60 min. Oxidation of the emulsion was monitored spectrophotometrically by measuring absorbance at 470 nm. Control sample contained 0.1 mL water. The degradation rate of -carotene was calculated by first order kinetics. Antioxidant activity (AOA) was expressed as percentage inhibition of -carotene degradation relative to the control after 60 min incubation, using the equation: AOA = 100 (DRc DRs ) (DRc)-1 where: AOA is antioxidant activity; DRc is degradation rate of the control, DRc = ln(ac /bc )/60; DRs is degradation rate of sample, DRs = ln(as /bs )/60; ln is natural log; a is absorbance at time 0; b is absorbance at time 60 min. Free radical scavenging activity of tomato extracts was determined by measurement of absorbance of 0.025 gL-1 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical solution with 0.1-0.5 mL tomato extract. Changes in absorbances at 515 nm were recorded at 10 sec intervals on Semco S/Ec spectrophotometer until the absorbance reached plateau. The percentage of remaining DPPH was calculated from the following equation: % DPPHrem = 100AT A0-1 where AT is an absorbance at time T needed to reach the steady state (beginning of plateau); A0 is an initial absorbance. The graph of the relation between % DPPHrem and phenolic compounds concentration of extract was then used to calculate from the equation of linear regression the amount of phenolic antioxi-
dants necessary to decrease the initial DPPH concentration by 50% (EC50 parameter). For easier and more clear presentation of results, the antioxidant power (AP) defined as the reciprocal of EC50 was used. AP = (EC50)-1 is expressed in miligrams DPPH per 1 mg phenols. Time [T(EC50)] needed to reach the steady state at EC50 was calculated graphically. The parameter antiradical efficiency (AE) which combines both EC50 and T(EC50) was determined from the following equation (Sanchez-Moreno et al. 1998): AE = [EC50 T(EC50) ]-1. AE is expressed in miligrams DPPH per 1 mg phenols and 1 min. Computer programme Excel 97 was used for calculation of kinetic parameters graphically. All data were subjected to one way analysis of variance and Newman-Keuls comparisons were carried out to test for significant (P=0.95) differences between means. RESULTS AND DISCUSSION Heating showed a significant loss of ascorbic acid (AA) content in the pulp. AA content declined with the increase of heating time (Table 1).
Table 1. Influence of heating conditions on the antioxidant constituents content of tomato pulps Ascorbic acid Lycopene Total flavocontent content noids content (mgkg-1f.w.) (mgkg-1f.w.) (mgkg-1f.w.) 160.6 a 145.0 b 142.7 b 16.8 c 23.2 b 34.3 a 26.5 b 44.1 a 20.8 c Total soluble phenolics content (mgkg-1f.w.) 181 d 327 a 226 b
Treatment Tomato pulp cold break processing Tomato pulp hot break processing Tomato pulp (cold break processing) heated at 90C for 2 min. Tomato pulp (cold break processing) heated at 90C for 10 min. Tomato pulp (cold break processing) heated at 90C for 20 min.
123.8 c
35.8 a
21.4 c
206 c
112.5 d
32.5 a
23.4 c
231 b
Note: Data in columns followed by the different letters differ significantly at P=0.95 using Newman-Keuls test
After 2, 10, 20 min of heating, AA level in the pulp diminished by about 11%, 23% and 30% respectively. Hot break processing only slightly decreased AA level of pulp (9.7% reduction) in comparison with cold break processing. Hot break processing significantly increased lycopene and total flavonoids contents in pulp by about 38% and 66% respectively in comparison with cold
break processing. Heating of cold break pulp at 90C markedly contributed to the increase lycopene content and slightly decreased flavonoids content. Lycopene content of heated pulp was about 2 fold higher than that from the cold break and its level remained stable apart from time of heating (Table 1). Thermal processing induced an increase of total soluble phenolics content (Table 1). Hot break processing increased total soluble phenolics content in the pulp by about 81% in comparison with cold break processing. After heating of cold break processed tomatoes pulp at 90C significant increase of soluble phenolics content was observed; the elevation of phenolics was 22% an average. Many papers have been reported that the thermal processing of tomatoes has detrimental effect on their ascorbic acid content (Dewanto et al. 2002, Sanchez-Moreno et al. 2006, Toor & Savage 2006). Losses of ascorbic acid content in tomatoes after heat processing at lower temperature (42-90C) were similar to our findings. The temperature and time at which tomato products are heated in the presence of air are the most important factors affecting the rate of ascorbic acid destruction (Gould 1983). As mentioned above (see Introduction) thermal processing of tomatoes increased the pool o bioaccesible lycopene. Our results clearly demonstrate that lycopene content increased in heat processed tomatoes. It is in agreement with the results of Dewanto et al. (2002) and Takeoka et al. (2001), however significant losses in lycopene content ranged 9%-28% during processing of juice into final paste using higher temperature conditions were also observed. Several publications mention inconsistent results for the behaviour of total phenolic compounds during thermal processing of tomatoes. The losses of the total phenolics content during thermal processing were reported by Kerkhofs (2003) (cited by Toor & Savage 2006) and Toor & Savage (2006) while Lavelli et al. (1999) and Gahler et al. (2003) observed an increase in the phenolics content. Dewanto et al. (2002) reported that heating of tomato pulp at 88C for 30 min did not affect the phenolics content of tomatoes. Our results clearly demonstrate that the total phenolics and flavonoids content in pulp significantly increased in hot break processed tomatoes and heating of pulp from cold break processed tomatoes enhanced total phenolics content. Since fruits and vegetables appear to have mixture of various antioxidants, some authors (Chu et al. 2000, Cao et al. 1996) recommend that antioxidant properties of vegetables should be evaluated by different methods rather than depending on the results of a single method. Therefore two different methods were chosen for the determination of antioxidant properties of tomatoes: scavenging the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and -carotene bleaching method based on coupled oxidation of -carotene and linoleic acid. The results of influence of heating on antioxidant properties of tomato pulp are presented in Table 2. Hot break processing considerably diminished antioxidant activity (AOA) of pulp by 63% and significantly increased antioxidant power (EC50)-1 by 47% in comparison with pulp from the cold break processing. Heating of tomato pulp from cold break processing diminished significantly highest value of AOA by 8% (at 2 min heating) and by 29% (at 10 min and 20 min heating). Value of (EC50)-1 of pulp from cold break processed tomatoes
heated at 90C for 2 min, 10 min and 20 min remained stable. The highest values of antiradical efficiency (AE) were found in pulp from cold and hot break processed tomatoes. AE expresses kinetic behaviour of antioxidants in free radical (DPPH) scavenging. Heating of pulp from cold break processed tomatoes decreased AE value by about 27% and it remained stable apart from time of heating. It has been observed that heat processed tomato juice had higher antioxidant activity than fresh tomatoes (Wang et al. 1996, Takeoka et al. 2001, Dewanto et al. 2002). Reasons for the increase in activity are unknown, but it may be at least partially explained by the production of new antioxidants during processing (Stewart et al. 2000). On the contrary, Toor & Savage (2006) found that the total antioxidant activity of the semi-dried tomatoes was significantly lower than that of the fresh samples.
Table 2. Influence of heating conditions on the antioxidant parameters of tomato pulps Antioxidant activity (AOA) (% inhibition of -carotene oxidation) 44.1 a 16.5 d 40.5 b Antioxidant power (EC50)-1 (mg DPPHmg-1 phenols) 25.2 a 37.1 a 21.3 b Antiradical efficiency (AE) [mg DPPH(mg phenolsmin)-1] 3.82 a 3.94 a 2.80 b
Treatment
Tomato pulp cold break processing Tomato pulp hot break processing Tomato pulp (cold break processing) heated at 90C for 2 min. Tomato pulp (cold break processing) heated at 90C for 10 min. Tomato pulp (cold break processing) heated at 90C for 20 min. Note see Table 1.
31.3 c
22.2 b
2.75 b
31.7 c
23.9 b
3.11 b
Regression analysis demonstrated high negative correlations between phenolics content and antioxidant activity (AOA) (r = -0.851, P = 0.99, Figure 1) and between total flavonoids and AOA (r = -0.74, P = 0.95, Figure 2) of tomato pulp. The statistical analysis also showed high positive correlations between phenolics content and antioxidant power (EC50)-1 (r = 0.816, P = 0.99, Figure 3) and between total flavonoids and (EC50)-1 (r = 0.935, P = 0.99, Figure 4).
50
45
40
35
30
AOA (%)
r2 = 0,7236 25
20
15
10
0 0 50 100 150 200 . 250 -1 300 350 400
Phenolics content (mg kg )
Fig. 1. Relationship between phenolics content and antioxidant activity (AOA) of tomato pulps
50 45 40 35
r = 0,5475
AOA (%)
30 25 20 15 10 5 0 0 10 20 30 . 40 -1 50
Total flavonoids content (mg kg )
Fig. 2. Relationship between total flavonoids content and antioxidant activity (AOA) of tomato pulps
45
40
35
Antioxidant power
30
r2 = 0,66 58
25
20
15
10
0 0 50 100 150 200 2 50 . -1 3 00 3 50 4 00
P he no lics co nte nt (m g kg )
Fig. 3. Relationship between phenolics content and antioxidant power (EC50)-1 of tomato pulps
45 40 35
Antioxidant power
30 25 20 15 10 5 0 0 5 10 15 20 25 30 35 40 45 50 r 2 = 0,8693
Total flavonoids content (mg/kg)
Fig. 4. Relatonship between total flavonoids content and antioxidant power (EC50)-1 of tomato pulps
Several studies have reported relations between phenolics content and antioxidant capacity. Toor et al.(2005) found in tomatoes that the phenolics (r =
0.84) and flavonoids (r = 0.88) were significantly related to the antioxidant activity (measured with ABTS radical decolourization assay) which suggests that measurement of total phenolics or flavonoids can be used as indicators of the antioxidant activity of tomatoes. Strong positive correlation between phenolics content and antioxidant activity in selected vegetables, fruits and grain products were reported (Velioglu et al. 1998, Kaur & Kapoor 2002). There is a wide degree of variation between different phenolic compounds in their effectiveness as antioxidants (Robards et al. 1999). The efficiency of phenolic compounds as antiradicals and antioxidants is diverse and depends on many factors, such as the number of hydroxyl groups bonded to the aromatic ring, their mutual position and the site of bonding (Sroka & Cisowski 2003). Some compounds which have DPPH scavenging activity may not show -carotene-linoleic acid antioxidant activity, relationship between these two models may not be obvious even for the same biological samples, that may contain a variety of antioxidants and pro-oxidants and this can be the result of synergies or antagonisms still unknown (Garcia-Alonso et al. 2004). Percentage of dry matter of freeze dried tomato slices was 6.03 0.153. Percentage of weight of baked tomato slices in relation to fresh ones was: 38.7 0.4 (for 200C, 1h) and 44.1 0.8 ( for 250C, 40 min). The results of influence of freeze drying and baking of tomato slices on antioxidant constituents are presented in Table 3. The results are calculated on the fresh weight basis, since the trends did not change after calculation on dry weight basis. Freeze drying of tomato slices contributed to the losses of ascorbic acid (16%) and phenolics content (21%). Most pronounced effect occurred for lycopene content. Freeze drying diminished lycopene level by 76 %, significantly decreased antioxidant power (EC50)-1 by 18% and antiradical efficiency by 29% (Table 4). Antioxidant activity (AOA) of freeze dried tomato slices was reduced to the level of 9.4% in comparison with this of raw tomato slices being 26.7% (Figure 5).
Table 3. Influence of freeze drying and baking conditions on the antioxidant constituents content of tomato slices Ascorbic acid Lycopene Total flavocontent content noids content (mgkg-1f.w.) (mgkg-1f.w.) (mgkg-1f.w.) 177.3 a 149.8 b 82.8 d 112.1 c 44.6 a 10.6 d 19.5 c 25.3 b 22.6 b 23.6 b 24.5 b 31.5 a Total soluble phenolics content (mgkg-1f.w.) 267 a 212 c 201 c 239 b
Treatment Raw tomato slices Freeze dried tomato slices Tomato slices baked at 200C for 1h Tomato slices baked at 250C for 40 min Note see Table 1.
Table 4. Influence of freeze drying and baking conditions on the antioxidant parameters of tomato slices Antioxidant power Antiradical efficiency (AE) (EC50)-1 (mg DPPHmg-1 phenols) [mg DPPH(mg phenolsmin)-1] 38.6 a 31.6 b 13.8 c 13.4 c 4.5 a 3.2 b 1.8 c 1.6 c
Treatment Raw tomato slices Freeze dried tomato slices Tomato slices baked at 200C for 1h Tomato slices baked at 250C for 40 min Note see Table 1.
30
a
25
20
AOA (%)
15
c
10
0 1 2 3 4
Treatments
Treatments: 1. Raw tomato slices; 2. Freeze dried tomato slices; 3. Baked tomato slices (200C, 1h); 4. Baked tomato slices (250 oC, 40 min) Note. Bars labelled by the different letters differ significantly at P=0.95 using Newman Keuls test Fig. 5. Antioxidant activity of phenolic extracts of fresh, freeze dried and baked tomato slices
Inspection of data presented in Table 3, 4 and in Figure 5 shows that baking of tomato slices contributed to the reduction of content of all determined components and of values of antioxidant parameters in comparison with raw tomato slices. However, the retention of ascorbic acid, lycopene, flavonoids and total soluble phenolic compounds in baked tomato slices was significantly higher at 250C, 40 min thermal treatment than that at 200C, 1h treatment. Little information exists about influence of freeze drying and baking on antioxidant properties and components of tomatoes. According to Morningstar et al. (2003) lycopene losses in freeze dried tomatoes ranged 20-40%. Seybold et al.(2004) found that baking of tomato slices (180-200C for 15-45 min) led to a significant rise of -tocopherol content. It seems to us that during baking, formation of novel compounds having anti- and/or pro-oxidant activity (i.e. Maillard reaction products) and decline of natural antioxidants (phenolics, ascorbic acid etc.) and the interactions among these different components had a crucial effect on the overall antioxidant properties of thermal processed tomatoes.
REFERENCES AOAC 1984. Official methods of analysis. (14th ed.), 43.064 43.068. Brand-Williams W., Cuvelier M.E., Berset C. 1995. Use of a free radical method to evaluate antioxidant activity. Lebensm.-Wiss. Technol. 26: 25-30. Cao G., Srfie F., Prior R.L. 1996. Antioxidant capacity of tea and common vegetables. J. Agric. Food Chem. 44: 3425-3431. Chu Y.H., Chang C.L., Hsu H.F. 2000. Flavonoid content of several vegetables and their antioxidant activity. J. Sci. Food Agric. 80: 561-566. Czapski J., Saniewski M. 1985. Effect of methyl jasmonate on carotenoids in tomato fruits. Gartenbauwissenshaft 50: 35-37. Davies B.H. 1965. Analysis of carotenoid pigments. In: Chemistry and biochemistry of plant pigments. Ed. T. Goodvin, Academic Press, London, New York, 489-532. Dewanto V., Wu X.Z., Adom K.K, Liu R.H. 2002. Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. J. Agric. Food Chem. 50: 3010-3014. Eberhardt M.V., Lee C.Y., Liu R.H. 2000. Antioxidant activity of fresh apples. Nature 405: 903-904. Emmons C.L., Peterson D.M., Paul G.L. 1999. Antioxidant capacity of oat (Avena sativa L.) extracts. 2. In vitro antioxidant activity and contents of phenolic and tocol antioxidants. J. Agric. Food Chem. 47: 4894-4898. Gahler S., Otto K., Bohm W. 2003. Alterations of vitamin C, total phenolics and antioxidant capacity as affected by processing tomatoes to different products. J. Agric. Food Chem. 51: 7962-7968. Garcia-Alonso M., dePascual-Teresa S., Santos-Buelga C., Rivas-Gonzalo J.C. 2004. Evaluation of the antioxidant properties of fruits. Food Chem. 84: 13-18. Gartner C., Stahl W., Sies H. 1997. Lycopene is more bioavailable from tomato paste than from fresh tomatoes. Am. J. Clin. Nutr. 66: 116-122. Giovanelli G., Zanoni B., Lavelli V., Niani R. 2002. Water sorption, drying and antioxidant properties of dried tomato products. J. Food Engin. 52: 135-141. Gould W.A. 1983. Tomato production, processing and quality evaluation (2nd ed.). Westport CT: AVI Publishing Company, Inc.
Kaur C., Kapoor H.C. 2002. Antioxidant activity and total phenolic content of some Asian vegetables. Int. J. Food Sci. Technol. 37: 153-161. Lavelli V., Hippeli S., Peri C., Elstner E.F. 1999. Evaluation of radical scavenging activity of fresh and air dried tomatoes by three model reactions. J. Agric. Food Chem. 47: 3826-3831. Mc Carthy M.A., Mattheus R.H. 1994. Nutritional quality of fruits and vegetables subjected to minimal processes. In: Minimally processed refrigerated fruits and vegetables. Wiley R.C. ed. Chapman and Hall, New York, 313-326. Morningstar D., Perkins-Veazie P., Rice S. 2003. Lycopene extraction from freeze dried tomatoes. Botany Conference, Mobile Alabama, July 26-31, Abstract ID: 480. Murcia M.A., Lopez-Ayerra B., Martinez-Tome M., Vera A.M., Garcia-Carmona F. 2000. Evolution of ascorbic acid and peroxidase during industrial processing of broccoli. J. Sci Food Agric. 80: 1882-1886. Nicoli M.C. Anese M., Parpinel M. 1999. Influence of processing on the antioxidant properties of fruit and vegetables. Trends Food Sci. Technol. 10: 94-100. Ragazzi E., Veronese G. 1973. Quantitative analysis of phenolic compounds after thin layer chromatographic separation. J. Chromatogr. 77: 369-375. Robards K., Prenzler P.D., Tucker G., Swatsitang P., Glover W. 1999. Phenolic compounds and their role in oxidative processes in fruits. Food Chem. 66: 401-436. Sanchez-Moreno C., Larrauri J.A., Saura-Calixto F. 1998. A procedure to measure the antiradical efficiency of polyphenols. J. Sci. Food Agric. 76: 270-276. Sanchez-Moreno C., Larrauri J.A., Saura-Calixto F. 1999. Free radical scavenging capacity of selected red, rose and white wines. J. Sci. Food Agric. 79: 1301-1304. Sanchez-Moreno C., Plaza L., de Ancos B., Cano M.P. 2006. Impact of high pressure and traditional thermal processing of tomato puree on carotenoids, vitamin C and antioxidant activity. J. Sci. Food Agric. 86: 171-179. Seybold C., Frohlich K., Bitsch R., Otto K., Bohm V. 2004. Changes in contents of carotenoids and vitamin E during tomato processing. J. Agric. Food Chem. 52: 7005-7010. Shi J., Le Maguer M. 2000. Lycopene in tomatoes: chemical and physical properties affected by food processing. Crit. Rev. Food Sci. Nutr. 40: 1-42. Sroka Z., Cisowski W. 2003. Hydrogen peroxide scavenging, antioxidant and antiradical activity of some phenolic acids. Food Chem. Toxicol. 41: 753-758. Stahl W., Sies H. 1992. Uptake of lycopene and its geometrical isomers is greater from heat processed than from unprocessed tomato juice in humans. J. Nutr. 122: 21612166. Stewart A.J., Bozonnet S., Mullen W., Jenkins G.I., Lean M.E., Crozier A. 2000. Occurrence of flavonols in tomatoes and tomato-based products. J. Agric. Food Chem. 48: 2663-2669. Takeoka G.R., Dao L., Flessa S., Gillespie D.M., Jewell W.T., Huebner B., Bertow D., Ebeler S.E. 2001. Processing effects on lycopene content and antioxidant activity of tomatoes. J Agric. Food Chem. 49: 3713-3717. Toor R.K., Savage G.P. 2006. Effect of semi-drying on the antioxidant components of tomatoes. Food Chem. 94: 90-97. Toor R.K., Lister C.E., Savage G.P. 2005. Antioxidant activity of New Zealand grown tomatoes. Int. J. Food Sci. Nutr. 56: 597-605. Velioglu Y.S., Mozza G., Gao L., Oomah B.D. 1998. Antioxidant activity and phenolics in selected fruits, vegetables and grain products. J. Agric. Food Chem. 46: 41134117.
Wang H., Cao G., Prior R.L. 1996. Total antioxidant capacity of fruits. J. Agric. Food Chem. 44: 701-705. Zanoni B., Peri C., Niani R., Lavelli V. 1999. Oxidative heat damage of tomato halves as affected by drying. Food Res. Int. 31: 395-401. Zhishen J., Mengcheng T., Jianming W. 1999. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 64: 555-559.
WPYW TERMICZNEGO PRZETWARZANIA POMIDORW NA WACIWOCI I SKADNIKI ANTYOKSYDACYJNE PRODUKTW POMIDOROWYCH
Streszczenie Badano wpyw zimnego i gorcego procesu przygotowania pulpy pomidorowej, ogrzewania pulpy oraz liofilizacji i pieczenia krojonych w plastry pomidorw na waciwoci i skadniki antyoksydacyjne otrzymanych produktw. Procesy termiczne obniyy zawarto kwasu askorbinowego oraz istotnie podwyszyy zawarto likopenu i fenoli rozpuszczalnych pulpy. Proces termiczny przygotowania pulpy pomidorowej znaczco obniy aktywno antyoksydacyjn (AOA) (oznaczon metod hamowania autooksydacji -karotenu w ukadzie -karoten kwas linolowy) o 63% i istotnie podwyszy potencja antyoksydacyjny (EC50)-1 (oznaczony metod zmiatania wolnego rodnika 2,2-difenyl-1-pikrylhydrazyl (DPPH) o 47%. Stwierdzono ujemn korelacj midzy zawartoci fenoli i AOA (r = -0,851) oraz zawartoci flawonoidw i AOA (r = -0,74). Stwierdzono rwnie wysok dodatni korelacj midzy zawartoci fenoli i potencjaem antyoksydacyjnym (EC50)-1 (r = 0,816) oraz zawartoci flawonoidw i (EC50)-1 (r = 0,935). Liofilizacja i pieczenie plastrw pomidorowych spowodoway istotne obnienie zawartoci wszystkich badanych skadnikw i wartoci parametrw antyoksydacyjnych w porwnaniu do wieych plastrw pomidorw.

THE STUDIES ON NEW TECHNOLOGIES FOR STORAGE PROLONGATION AND MAINTAINING THE HIGH QUALITY OF VEGETABLES
Franciszek ADAMICKI, Ewa BADEEK Research Institute of Vegetable Crops Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland
Summary The investigations were undertaken to study the effect of new technologies of vegetable storage in ultra low oxygen (ULO) atmosphere, Modified Interactive Packaging (MIP) and 1-MCP treatment on storage and quality maintaining of Chinese cabbage, broccoli, lettuce (crisphead) and tomatoes. Two cultivars of Chinese cabbage Bilko F1 and Morillo F1 were stored under following CA condition: 2.5% CO2 - 1.5% O2 and 1% CO2 - 1% O2, and control (air) at temperature 0C for 130-150 days. Lettuce and broccoli were stored in MIP packages as well as in plastic boxes lined with polyethylene film (PE) at 0C respectively for 4 and 9 weeks. Mature green tomato fruits cv. Faustine F1, grown in the field, treated with of 0.5 LL-1 and 1.0 LL-1 1-methylcyclopropene for 21 hours at 18C were stored at 12.5 and 20C and 95% relative humidity. There was found different reaction of compared two Chinese cabbage cultivars for ULO atmosphere storage. Significantly higher percentage of marketable heads (66-82%) in case of cv. Morillo F1 compared to cv. Bilko F1 respectively 58-80% was noted. Modified atmosphere inside of MIP have beneficial effect on retention of green colour of heads, their firmness, quality and weight loss of broccoli. Broccoli heads stored with one layer of leaves gave a little better results than without leaves. There werent found any significant differences in quality evaluation parameters of iceberg lettuce stored in MIP packages or in boxes lined with polyethylene film, however butt discoloration, rotting of leaves as well as marketable quality seems to be better in MIP. Treatments with 1-MCP resulted in delay ripening of mature green tomatoes. key words: Chinese cabbage, lettuce, broccoli, tomato, ULO, 1-MCP treatment, MIP packaging INTRODUCTION Many species of green vegetables such as crisphead lettuce, broccoli, Chinese cabbage are highly perishable and being subject to high quality loss during storage and marketing. The newest technologies - controlled atmosphere, in-
cluding ULO and modified atmosphere packaging (MAP, MIP and others) are now being increasingly used in the practice for extending storage life and prolonging product quality and reducing the loss of a wide range of fresh stored vegetables (Adamicki 2003). Storage of Chinese cabbage in controlled atmosphere, as well as in ULO, has been increased in the last few years in Poland to volume around 1000 ton. Use of controlled atmosphere allows to reduce loss in compared to traditional methods of storage and maintain quality and nutritional value of produce. However low concentration of CO2 and O2 delays yellowing of the leaves and reduce decay of heads, some physiological disorder are still the main problem which should be clarified. Modified atmosphere packaging (MAP) is widely used for prolonging storage period of vegetables due to decreasing O2 and increasing CO2 inside the package atmosphere obtained mainly by respiration activity of stored produce (Artes & Martinez 1996, Day 1998, Adamicki 2001, Singh & Sudhakar 2005). The newest solution in packaging developments of modified interactive packaging (MIP) work in a totally different manner. Packaging are made from low density polyurethane impregnated with mineral compounds that enable acquirement unique film with selective gas permeation (Anon 2000). This allows for a set percentage of oxygen to enter the bag at any given time. Inside the bags maintain the high levels of humidity with minimal condensation. The main advantage of the MIP packages is its wide spectrum of usage, e.g. the same bags can be used for storage of vegetables, fruits, herbs and other fresh products. Concentration of CO2 inside MIP packages remains near constant during whole period of storage and level depends on kind of stored produce, e.g. for broccoli reached 3.5 till 4%, for crisphead lettuce 1.5 till 2.2% and tomatoes 2 till 3% (Adamicki 2001). The concentration of O2 inside MIP packages decreases rapidly on the beginning of storage period and then increases slowly up to level of 6 till 12.5% upon the vegetable (Anon 2000, Adamicki 2001). Packaging of stored broccoli with different types of film was the most effective treatment for reducing of weight loss, maintaining chlorophyll content, sensory and nutritional quality (Serrano et al. 2006). Tomatoes represent one of the most important source of vitamins and minerals for human diet, since they are frequently consumed uncooked. Prior and post harvest procedures, inappropriate storage condition and shelf-life can affect the quality and nutritive value of tomatoes (Horbowicz & Saniewski 2000, Guillen et al. 2005, Sablani et al. 2006). Controlled atmosphere is one of the storage method for mature green tomatoes used in the practice to delay ripening and maintain quality after ripening (Adamicki & Magomedov 2004). The discovery of 1-MCP as an ethylene inhibitor in the late 1980 and a considerable information collected throughout the last 20 years seem to be sufficient to use in practice for extending the life of many fruit and vegetables (Bower & Mitcham 2001, Prange & DeLong 2003, Guillen et al. 2005). 1-MCP can be useful to extend storage life of mature green tomato fruit while still allowing normal ripening after some period of storage (Bower & Mitcham 2001, Adamicki & Badelek 2003). The commercial advantage of use of 1-MCP for tomatoes is similar to controlled atmosphere storage with much lower costs of
F. ADAMICKI, E. BADEEK THE STUDIES OF NEW TECHNOLOGIES ... 65 _____________________________________________________________________________________________________
investment. From this point of view 1-MCP becomes a new tool with a great potential for use in practice in near future. The quality and nutritional value of vegetables is affected by postharvest handling and storage conditions. Tomato, crisphead lettuce and broccoli are widely consumed vegetables in Poland and are very important sources of vitamins and other health promoting components as lycopene, carotene, sulforaphane and phenols. Some of them are antioxidant, which reduce the risk of arteriosclerosis, cardiovascular diseases and some form of cancer (Horbowicz & Saniewski 2000, Sablani et al. 2006). The objective of the study was to evaluate the effect a new technology system, including controlled atmosphere, 1-MCP treatments and MIP packaging, on the storage life, quality and nutritional value of some vegetable species. MATERIAL AND METHODS Chinese cabbage cv. Morillo Storido F1 and Bilko F1, broccoli cv. Marathon F1, and crisphead lettuce cv. Balance and Bergland were obtained from experimental field of the Research Institute of Vegetable Crops, Skierniewice. Mature green tomato fruits cv. Faustine F1 were harvested at the end of August from producer field where plants were grown with stacks under good management practice. Just after harvest Chinese cabbage were placed into 500 L gas tight containers, which were closed and set to required composition of the atmosphere. Each treatment consisted of 4 replication, containing of 6-8 heads of cabbage. During storage the required controlled atmosphere maintained and corrected automatically by Oxystat 200 System, David Bishop Inst. Oxygen and carbon dioxide concentration were maintained in the range of 0.2% from the fixed level. Chinese cabbage was stored at 0 and 95-98% RH for 130 and 150 days. Fresh harvested mature green tomato fruit before storage has been washed in chlorinated water (300 ppm calcium hypochlorite) for 30 seconds. Then fruits were air dried to remove the excess of surface moisture and placed in gas tight containers for 1-MCP treatment. For 1-MCP treatment of tomatoes SmartFreshTM as a powder was diluted in water and released 1-MCP as a gas. Tomatoes were kept during 21 hours at temperature 18C at 1-MCP concentration of 0.5 LL-1 and 1.0 LL-1. After treatments all fruit were stored up to 10 weeks at 12.5 or 20 in the dark. For each treatment and replicate, evaluation of visual quality and ripening progress were made twice weekly. Harvested broccoli and crisphead lettuce, after initial cooling to storage temperature, were packed in the modified interactive packaging (MIP bags). Bag sizes were adopted to 20 kg plastic boxes. In the control treatment the vegetables were stored in similar boxes, lined with 35m polyethylene film that enabled free air circulation with the outer atmosphere. The broccoli and crisphead lettuce were kept at 0C. The gas composition (% of carbon dioxide) inside the MIP packs was monitored at 2 days intervals with ANAGAS CD-98 analyzer, while % of oxygen was analyzed twice a week with Hewlett-Packard HP 5890 gas chromatograph. After storage, the weight loss and marketable
value were evaluated, as well as other traits concerning vegetable quality (colour, firmness and compactness of broccoli heads, discoloration of crisphead lettuce) according to score scale used in Laboratory of Storage and Postharvest Physiology (10 excellent, 8 good, 6 satisfactory (acceptable for market), 3 poor (non edible vegetables). The experimental design was completely randomised. The obtained results were statistically analysed by ANOVA and the significance among treatment mean values were compared with Newman-Keuls test at the P=0.05 level. RESULTS AND DISCUSSION Chinese cabbage Low concentration of CO2 and O2 seems to be one of the most effective method in extending the storage life of Chinese cabbage. Percentage of marketable heads of Chinese cabbage stored in ultra low concentration of oxygen and carbon dioxide depends on cultivar, storage period and vegetation season. Among two compared cultivars better results were obtained for Morillo Storido F1 than Bilko F1 (Table 1 & 2). Statistically significant higher percentage of marketable heads for cv. Morillo Storido F1 was noted during three years of experiments in controlled atmosphere than in air. The better concentration for cv. Bilko F1 seems to be ultra low atmosphere containing only 1.0% oxygen and carbon dioxide. Physiological disorders of cabbage heads (black small spots on the midribs and brown midribs) were observed occasionally during storage in controlled atmosphere and regularly for cv. Morillo Storido F1 in air at all three seasons.
Table 1. Effect of controlled atmosphere (ULO) on the storage and quality of Chinese cabbage (temp.0C; cv. Morillo Storido F1; in % of initial weight) Storage conditions Control (air) 2000/01 2.5% CO2 - 1.5% O2 1.0% CO2 - 1.0% O2 Control (air) 2001/02 2.5% CO2 - 1.5% O2 1.0% CO2 - 1.0% O2 Control (air) 2002/03 2.5% CO2 - 1.5% O2 1.0% CO2 - 1.0% O2 Years Marketable heads 70.4 a 82.0 a 80.8 a 4.7 b 72.0 a 66.7 a 37.5 b 71.3 a 70.3 a Physiological damage 10.7 0 0 91.9 b 4.9 a 10.9 a 37.0 0 0 Total loss 29.6 b 18.0 a 19.2 a 95.3 b 28.0 a 33.3 a 62.5 b 28.7 a 29.7 a
Note: Storage period: 2000/01 130 days; 2001/02 and 2002/03 150 days Means followed by the same letter do not differ significantly at P=0.05, Newman-Keuls test
Table 2. Effect of controlled atmosphere (ULO) on the storage and quality of Chinese cabbage (temp.0C; cv. Bilko F1; in % of initial weight) Storage conditions Control (air) 2000/01 2.5% CO2 - 1.5% O2 1.0% CO2 - 1.0% O2 Control (air) 2001/02 2.5 % CO2 1.5 % O2 1.0 % CO2 1.0 % O2 Control (air) 2002/03 2.5 % CO2 1.5 % O2 1.0 % CO2 1.0 % O2 Years Note: see Table 1 Marketable heads 71.4 a 75.7 a 80.3 a 74.1 a 59.1 b 63.0 ab 69.9 ab 58.8 b 79.6 a Physiological damage 13.9 0 0 0 0 9.7 0 0 0 Total loss 28.6 b 24.3 a 19.7 a 25.9 a 40.9 b 37.0 ab 30.1 ab 41.2 b 20.4 a
Obtained results are in agreement with previous experiment conducted with different cultivars of Chinese cabbage stored in CA and air (Adamicki 2003). The conclusion from this study is that storage ability of two cultivar of Chinese cabbage is greatly influenced by the seasonal changes. It was demonstrated by significant differences in amount of marketable heads and total loss for both compared cultivars. Obtained results can be explained by specific climate conditions just before harvest. Storage in controlled atmosphere significantly reduce physiological damage of Chinese cabbage but especially cv. Morillo Storido F1, but also depends from storage season. Broccoli The wrapping of broccoli heads with films, which generate a modified atmosphere is a good tool to provide a optimal storage conditions. Concentration of CO2 inside the modified interactive packages (MIP) after two days of storage of broccoli at 0C increased up to 5.6 to 5.9% and then slowly decreased reaching after 16 days to 3.5-3.7% and kept on this level near to the end of the storage (Fig. 1). At the same time concentration of O2 inside the packages sharply decreased from 21 to 3.5% after 11 days of storage and then kept on this level, with small fluctuation, throughout period of storage (data not shown on figures). Modified atmosphere inside packages provide a good storage conditions for preventing of broccoli yellowing and allow to maintaining excellent quality of heads (Table 3). However there wasnt found a statistically significant differences between marketable ability and quality of broccoli stored in MIP packages and in boxes lined with polyethylene film. The amount of marketable broccoli, quality evaluation and colour of heads seems to be a little better in modified interactive packages in comparison to control. Modified interactive packages are very simply for using in the practice and allow to prolong storage of broccoli in same cases even up to 3 months. This method is much cheaper than storage in controlled atmosphere and gave a better results in comparison to storage of broccoli in cold room. Modified atmosphere obtained
during storage of broccoli in MIP allow to maintains a green colour of heads, thereby reduces the loss of chlorophyll. Due to maintaining inside the packages a high level of relative humidity, the decrease of the weight loss and wilting of broccoli heads as well as reduction of decay of broccoli was observed.
7
MIP 1 MIP 2 MIP 3
% CO2
0 2 5 7 9 12 14 16 19 21 23 28 30 33 35 40 42 44 51 58
Storage period (days)
Fig. 1. Concentration of CO2 inside modified atmosphere packaging of broccoli (temp. 0C; cv. Marathon F1; 2004) Table 3. Effect of modified interactive packaging on the quality characteristics of broccoli (temp. 0C; cv. Marathon F1; storage period 9 weeks) Year At harvest 2003 2004 2005 Control MIP Control MIP Control MIP Control MIP Control MIP Control MIP Pre-packaging Quality Marketable heads (%) 10.0 10.0 Heads without leaves 6.9 8.7 68.7 7.8 8.9 86.7 8.5 8.5 100.0 8.5 8.8 100.0 8.8 9.2 93.3 9.2 9.4 97.8 Heads with leaves 7.7 b 8.9 a 80.7 b 8.5 a 9.1 a 100.0 a 8.2 a 8.7 a 91.7 b 8.8 a 8.9 a 100.0 a 7.6 b 9.7 a 72.2 b 9.3 a 9.3 b 100.0 a Colour Weight loss 2.3 b 1.9 a 1.3 a 1.4 a 1.5 b 1.0 a 2.0 a 1.6 a 2.0 b 1.4 a 2.9 b 1.3 a
2003 2004 2005
Means followed by the same letter do not differ significantly at P=0.05, Newman-Keuls test
Visual quality score: 10 - excellent, 8 good, 6 acceptable, 3 poor - non edible
The effect of modified atmosphere storage on reduction of weight loss, decrease of yellowing and quality deterioration has been reported by many authors (Toivonen 1997, Day 1998, Serrano et al. 2006). Presented results showed that storage of broccoli in modified atmosphere is one of the most effective treatments for reduction of weight loss, wilting and maintaining green colour and high quality of heads even after long period of storage. Crisphead lettuce Cold storage slows down the rate of respiration and ageing of crisphead lettuce, however using different packaging unit it could be possible to reduce further all processes by modify atmosphere around product. During storage of crisphead lettuce in modified interactive packages concentration of CO2 during first two days increased to 1.7-1.9% and remained on the same level throughout whole period of storage while in other packages increased up to 2.5% (Fig. 2). Concentration of O2 decreased after 4 weeks of storage to 9-12% (data not shown on figures). Weight loss of lettuce stored in MIP and in boxes lined with polyethylene film (control) did not exceed 0.8% and were a little bit lower in control treatments. Marketable value of lettuce head stored in MIP packages was evaluated as saleable and acceptable for market, while lettuce stored in polyethylene lined crates has worse quality grade (Table 4). On the lettuce heads stored in air and placed in MIP butt discoloration and midribs were the main postharvest disorders developed. According to our present results MIP packages has prolonged the storage life of lettuce in comparison to control treatment (lined boxes with PE film) but without any significant effect on decrease of discoloration, rotting and marketable quality of heads. Similar results has been also in a wide range of vegetables (Artes & Martinez 1996, Anon 2000). Butt discoloration was much higher than midribs in both treatments. There werent found any differences in discoloration of midribs and butts between methods of packaging.
2,5
1,5
% CO2
MIP 1 MIP 2 MIP 3
0,5
0 1 2 3 6 8 10 13 15 17 20 22 24
Storage period (days)
Fig. 2. Concentration of CO2 inside modified interactive packaging of crisphead lettuce (temp. 0C; storage period:6 July-6 Aug.2004; cv. Bergland) Table 4. Effect of modified interactive packaging on the quality of crisphead lettuce (temp. 0C; storage period 4 weeks) PreCompactness Discoloration packaging of head midribs butt At harvest 5.0 1.01 1.01 Control 3.9 2.7 5.6 Balance MIP 4.1 2.1 5.9 2003 Control 4.1 2.7 6.0 Bergland MIP 4.2 3.1 5.9 Control 4.4 2.5 5.9 Balance MIP 4.2 2.7 6.0 2004 Control 4.3 2.7 6.1 Bergland MIP 4.5 2.1 6.2 Year Cultivar
1
Rotting 1.01 3.2 b 1.3 a 2.0 a 2.4 a 2.6 a 2.9 a 2.7 a 2.1 a
Marketable quality 9.02 4.7 b 6.6 a 4.8 b 5.0 b 4.8 a 4.1 a 4.9 a 5.6 a
Means followed by the same letter do not differ significantly at P=0.05, Newman-Keuls test
1-none, 9-severe; 2 9-excellent, 1-unusable
Tomatoes Alternative method, to CA storage, which can be used for delay of tomato ripening and to prolong their storage life is 1-MCP treatment. Significant differences between 1-MCP treated and control (untreated) mature green fruits in ripening delay, e.g. number of days to reach breaker stage of maturity and number of days to reach table ripe stage has been found in our experiments (Table 5). In non treated tomato fruits ripening proceeds more quickly and fruits reach table ripe stage after 20.4 days at 12.5C, while 1-MCP treated fruits after 35.8
days. Quality score of ripe fruits were similar (8.9 points) even if the storage period for 1-MCP treated tomatoes was nearly two times longer.
Table 5. Effect of 1-MCP treatment and storage temperature on ripening and quality of mature green tomatoes (average from 2002-2003; Faustine F1) Treatment 1MCP concentration Control (untreated) 0.5LL-1 1.0 LL-1 Control (untreated) 0.5LL-1 1.0 LL-1 Storage temp. 12.5C Days to breaker Days to table ripe 8.0 c 20.4 b 13.4 b 30.1 a 17.0 a 35.8 a Storage temp. 20C 3.8 c 10.2 b 8.4 b 19.0 a 10.1 a 23.6 a
Quality 8.9 8.7 8.9 8.9 8.6 8.6
Visual quality score: 9 - excellent, 7 good, 5 acceptable, 3 poor - non edible Means followed by the same letter do not differ significantly at P=0.05, Newman-Keuls test
Higher concentration of 1-MCP, 1.0 LL-1, was more effective in ripening delay and in storage extending period of tomato fruits. Similar results has been obtained in previous reports (Adamicki & Badelek 2003, Guillen et al. 2005). Higher temperature of storage (20C) has significant influence on decrease the number of days to reach breaker stage and table ripe stage of tomatoes both non treated and treated with 1-MCP as well as quality evaluation. The present finding showed that 1-MCP could be a new tool with a great potential for use in the practice, instead of CA, to prolong storage and extend shelf-life of mature green tomatoes. This is consistent with the views of Prange & DeLong (2003) and Guillen et al. (2005).
REFERENCES Adamicki F. 2001.The effect of modified interactive packaging (MIP) on post-harvest storage of some vegetables. Veget. Crops Res. Bull. 54: 213-218. Adamicki F. 2003. Effect of Controlled Atmosphere and temperature on physiological disorders of stored Chinese Cabbage (Brassica rapa L. var. pekinensis). Acta Hort. 600: 297-301. Adamicki F., Badeek E. 2003. Effect of 1-MCP on ripening, quality and storage potential of tomatoes. Folia Hort. Supl. 2003/2: 361-363. Adamicki F., Magomedov R.K. 2004. Chranenie plodov tomatov v reguliruemoj gazovoj sredie. Vestnik Ros. Akad. Selskochoz. Nauk 5: 79-80. [in Russian] Anon 2000. Long Life Solution. Modified Interactive Packaging. Leaflet, p.24. Artes F., Martinez J.A. 1996. Influence of packaging treatments on the keeping quality of Salinas lettuce. Lebensmittel-Wissenschaft & Technologie, 29(7): 664-668. Bower J., Mitcham B.2001. Application of 1-MCP to vegetable crops. Perishable Handling Quarteerly 108: 26-27. Day B.P.F. 1998. Novel MAP a brand new approach. Food Manufacture 11: 22-24.
Guillen F., Valverde J.M., Martinez-Romero D., Castillo S., Valero D., Serrano M. 2005. Tomato fruit quality retention during storage by 1-MCP treatment as affected by cultivar and ripening stage at harvest. Acta Hort. 682: 1069-1075. Horbowicz M., Saniewski M. 2000. Biosynteza, wystpowanie i waciwoci biologiczne likopenu. Post. Nauk Roln. 1: 29-46. [in Polish with English summary] Prange R., DeLong M. 2003. 1-Methylcyclopropene: the magic bullet for horticultural products. Chronica Hort. 43: 11-14. Sablani S.S., Opara L.U., Al.-Balushi K. 2006. Influence of bruising and storage temperature on vitamin C content of tomato fruit. J. Food Agr. & Environ. 4(1):54-56. Serrano M., Martinez-Romero D., Guillen F., Castillo S., Valero D. 2006. Maintenance of broccoli quality and functional properties during cold storage as affected by modified atmosphere packaging. Postharvest Biol. Technol. 39: 61-68. Singh S.P., Sudhakar Rao D.V. 2005. Effect of modified atmosphere packaging (MAP) on the alleviation of chilling injury and dietary antioxidants levels in Solo papaya during low temperature storage. Europ. J. Hort. Sci. 70(5): 246-252. Toivonen P.M.A. 1997. The effects of storage temperature, storage duration, hydrocooling, and micro-perforated wrap on shelf-life of broccoli (Brassica oleracea L., Italica Group). Postharvest Biol. Technol. 10: 59-65.
BADANIA NAD NOWYMI TECHNOLOGIAMI POZWALAJCYMI NA PRZEDUENIE OKRESU PRZECHOWANIA I ZACHOWANIE WYSOKIEJ JAKOCI WARZYW Streszczenie Przeprowadzono dowiadczenia z wykorzystaniem nowych technologii, takich jak ultraniska koncentracja tlenu (ULO), opakowania interaktywne (Modified Interactive Packaging), 1-MCP i ich wpywu na trwao i jako przechowywanych warzyw kapusty pekiskiej, brokuw, saaty kruchej i pomidorw. Dwie odmiany kapusty pekiskiej Bilko F1 i Morillo F1 byy przechowywane w kontrolowanej atmosferze zawierajcej 2,5% CO2 1,5% O2 i 1% CO2 - 1% O2 oraz w normalnej atmosferze w temperaturze 0C przez okres 130-150 dni. Saata krucha i brokuy byy przechowywane w opakowaniach interaktywnych i w opakowaniach wykadanych foli polietylenow (kontrola) w temperaturze 0C przez okres 4-9 tygodni. Zielone, wyronite owoce pomidorw odmiany Faustine F1, pochodzce z uprawy polowej, traktowano 1-MCP (1metylocyklopropen) w steniu 0,5 L L-1 i 1,0 L L-1 w cigu 21 godzin w temperaturze 18C i nastpnie przechowywano w normalnej atmosferze w temperaturze 12,5C i 20C oraz wilgotnoci wzgldnej powietrza 95%. Stwierdzono rnice w trwaoci przechowalniczej odmian kapusty pekiskiej. Wyszy procentowy udzia gwek handlowych po okresie przechowania notowano dla odmiany Morillo F1 (66-82%) w porwnaniu do odmiany Bilko F1(58-80%). Zmodyfikowana atmosfera utrzymujca si wewntrz opakowa interaktywnych (MIP) miaa korzystny wpyw na zachowanie zielonego zabarwienia, jdrnoci, jakoci i ubytkw masy brokuw. Lepsze wyniki uzyskano, gdy re brokuw przechowywano z 1 okkiem lici okrywajcych. Nie stwierdzono istotnych rnic w jakoci saaty kruchej przechowywanej w opakowaniach MIP i w skrzynkach wykadanych foli polietylenow, aczkolwiek brzowienie ogonkw liciowych, gnicie i warto handlowa bya nieznacznie wysza po przechowaniu w opakowaniach interaktywnych. Pozbiorcze traktowanie owocw pomidorw 1-MCP hamowao ich wybarwianie si i znacznie przeduao okres przechowania zarwno w temperaturze 12,5C jak w 20C.

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THE INFLUENCE OF STORAGE ON SENSORY CHARACTERISTICS OF EGGPLANT FRUITS (SOLANUM MELONGENA L.) GROWN IN FOIL TUNNEL AND GREENHOUSE
Marek GAJEWSKI, Katarzyna KOWALCZYK, Daria ARASIMOWICZ Department of Vegetable and Medicinal Plants Warsaw Agricultural University Nowoursynowska 166, 02-787 Warszawa, Poland
Summary The aim of this work was to examine the effect of storage of eggplant fruits on their sensory characteristics. Fruits were harvested in the middle of September, at their optimal maturity, and then stored for three weeks at 12oC and 95% RH. Cultivars chosen for the experiment were Bibo and Long Black. Plants were grown in unheated foil tunnel and in greenhouse. Sensory quality was analyzed immediately after harvest and after storage, using the QDA method, by the panel of 12 trained experts. Fruits were roasted for 40 minutes at 180oC, and then chilled to the ambient temperature. The set of 19 descriptors of sensory quality plus overall quality were evaluated. Storage of fruits significantly affected some sensory descriptors. Fruits from the tunnel showed less intensive odor of steamed potatoes after storage, but more intensive odor of a plum jam. Flesh firmness, flesh fibrousness and skin hardness were lower after storage. During storage sweet taste intensity decreased and bitter taste intensity increased in all fruits. In case of fruits from the greenhouse storage affected odor intensity as well. Overall quality of all cultivars was scored lower after storage, however off-odor and off-flavor intensity were very low. Differences between cultivars grown in the foil tunnel were significant concerning sweet taste and bitter taste intensity, and in case of fruits grown in the greenhouse concerning flesh juiciness and skin hardness. key words: aubergine, cultivars, quantitative descriptive analysis, sensory quality, storage INTRODUCTION Eggplant (aubergine) is a cold sensitive plant, widely grown in Mediterranean and Far East regions. In Polish climatic condition it can be grown successfully in greenhouses or foil tunnels only (Gajewski & Gajc-Wolska 1998). However, popularity of this vegetable in Poland has increased during last few years. Eggplant fruits are harvested for consumption at a physiologically nonmature stage. They are a valuable component of human diet. In fresh weight
they contain about 7% of dry matter, 1% of proteins, and 4% of carbohydrates. Also vitamins B1, B2, B6 and C were found in fruits (Rohde 1976). Eating of eggplant fruits can decrease LDL level in human blood due to hypolipidemic effect of some flavonoids and improve heart action (Sudheesh et al. 1997, Kashyap et al. 2003). Eggplant fruits are chilling sensitive and critical temperature is 5oC. Therefore, the temperature of 10-12oC is recommended for storage (Abe & Ogata 1978, Aubert & Pochard 1981, Mencarelli et al. 1989, Gajewski 2000a, b, Jha & Matsuoka 2002). Esteban et al. (1992) found that during development of fruits titratable acidity, reducing and total sugars, ascorbic acid, proteins and total phenolics content increase. After 18 days storage of fruits in 10oC ascorbic acid content decreased by 75%. Total sugars decreased from 4 to 2%. Total polyphenolics content decreased, especially when fruits were kept in 20oC. Titratable acidity decreased initially and then increased, but pH increased from 4.5 to 5.2-5.4. Gajewski (2000 a, b) reported that during storage firmness of eggplant fruits and total sugars content decreased. It can be presumed that some of these changes may affect also sensory characteristics of stored fruits. Sensory evaluation of vegetables brings very valuable information on their quality characteristics. Sensory traits of vegetable are usually the main factor determining consumers satisfaction (Abbott 1999). Among various sensory evaluation methods reported in literature, the QDA method (quantitative descriptive analysis) is usually applied for detailed description of sensory characteristics. In this method an assumption is done that sensory quality is a complex of many descriptors, which can be individually estimated by a consumer (Meilgaard et al. 1999, Chabanet 2000). For the unification of sensory evaluation methods international standards were approved, based on ISO recommendations (Anonymous 1996, 1998, 1999). There are few reports concerning sensory characteristics of eggplant fruits. Gajewski & Arasimowicz (2004) found that eggplant cultivars grown in foil tunnel differ in some sensory traits when they were evaluated immediately after harvest and maturity stage of fruits also affects their sensory quality. The aim of this work was to examine if storage of eggplant fruits affects changes in their sensory characteristics. The other aim of the experiment was to determine if these changes are similar in case of fruits grown in a foil tunnel and in a greenhouse. An approach was also done to find out which sensory descriptors of eggplant fruits most significantly affect overall sensory impression during their consumption. MATERIAL AND METHODS The two-year experiment was carried out at the Warsaw Agricultural University. Two growing technologies of plants were applied: - in an unheated foil tunnel, in a natural soil (medium mud soil); - in a greenhouse, in the growing medium based on a peat. The two growing methods were applied since these are most common in eggplant cultivation in Poland. Eggplants were grown from transplants, planted
M. GAJEWSKI et al. THE INFLUENCE OF STORAGE ... 75 _____________________________________________________________________________________________________
on the final places in the middle of June (foil tunnel) or middle of July (greenhouse). The plants were trained, with three stems left, and tied to strings. Fertilizing was applied according to soil analysis results in the tunnel in solid form and in the greenhouse in a liquid form, using a drop irrigation system. Plants were chemically protected against pests and diseases. Air temperature was monitored. In the foil tunnel during fruit development (from the middle of August) the temperature in the foil tunnel varied from 25-30oC during the day to 15-20oC during the night. The temperature in the greenhouse varied from 2025oC during the day to about 18-20oC during the night. The fruits for sensory evaluation were harvested in the middle of September, at optimal maturity stage. Marketable quality fruits were then stored for three weeks in a cold store at the temperature of 12oC and 95% RH, packed in bags made of microperforated polyethylene film (three fruits in each bag). Factors of the experiment were: factor A: term of evaluation immediately after harvest, after 3-weeks storage at 12oC; factor B: cultivar. Two cultivars with different fruit shape and colour were chosen - Bibo F1 (creamy-white, oval shaped), Long Black (black-violet, elongated). Sensory evaluation was performed in sensory laboratory of the Department of Vegetable and Medicinal Plants. The trained panel consisting of 12 persons, previously selected and trained according to ISO standard (Anonymous 1996), carried out the sensory analysis. The assessment was carried out in a laboratory equipped according to ISO standard (Anonymous 1998). At the first part of QDA procedure brainstorming sessions were run to select sensory attributes for eggplant fruits. Panelists received samples of fruits varying in sensory properties and individually generated a set of 19 descriptors for odour, colour, texture and flavour. Also overall quality of fruits was scored (Table 1). For the evaluation of fruits the quantitative descriptive analysis (QDA) was used. Every assessor was given randomized samples of fruits. The analysis was performed in separate booths, equipped with computers for data acquisition. For the assessments whole fruits were roasted in foil bags for 40 minutes at the temperature of 180oC and then cooled to the ambient temperature. Samples of the fruits (slices thickness 1 cm) were put to coded plastic boxes, covered with lids, and then served to the assessors. The assessments were marked on non-structural lines, on the monitors. These lines showed also the anchoring points for each descriptor (low intensity high intensity). Results were converted to numerical values (from 0 to 10 units). The analysis was performed during two independent sessions, in two replications. For coding samples and for initial processing of the data Analsens software was used. For the analysis of variance Statgraphics Plus 4.1 software was applied, and LSD test was used to show which values differ significantly at P=0.05. Principal component analysis and regression analysis were also applied.
Table 1. Definitions of sensory descriptors used in the quantitative descriptive analysis

No. Odour None very intensive Characteristic odour of steamed potatoes with None very 2 Odour of steamed potatoes a skin intensive None very 3 Odour of boiled fungi Characteristic odour of boiled fresh fungi intensive None very 4 Odour of hay Characteristic odour of long stored hay intensive None very 5 Odour of a plum jam Sweet fruity odour, characteristic to a plum jam intensive Odour of boiled vegeta- Characteristic odour of boiled root vegetables None very 6 bles celeriac, carrot intensive 7 Off-odour Unusual odour for eggplant fruit Appearance Light dark 8 Flesh colour Visual evaluation of flesh colour brown NonFlesh visual evaluation in respect of develop9 Seeds development developed ment of seeds developed Texture 10 Flesh firmness Degree of force needed to chew the flesh Firm - soft Amount of liquid released when chewing Not juicy 11 Flesh juiciness a sample very juicy Smooth 12 Flesh fibrousness Mouthfeel of flesh homogenousness very fibrous 13 Skin hardness: Degree of force needed to bite the skin Hard soft Flavour / taste None very 14 Sweet taste Basic taste intensive None very 15 Flavour of boiled fungi Characteristic flavour of boiled fresh fungi intensive Characteristic flavour of roasted apples or None very 16 Flavour of a roasted fruit plums intensive None very 17 Bitter taste Basic taste intensive None very 18 Pungent flavour Gives an impression of burning on the tongue intensive None very 19 Off-flavour Unusual flavour for eggplant fruit intensive Low high Overall quality Score for general sensory quality impression quality 1 Sharp odour Pungent, spicy odour Descriptor Definition Anchoring points
RESULTS AND DISCUSSION Storage affected some of sensory descriptors of eggplant fruits, and these changes were slightly different in case of fruits from cultivars grown in the
plastic tunnel and in the greenhouse. The results are shown separately for the two growing method, based on the two years data. Fruits from the plastic tunnel showed less intensive odour of steamed potatoes after storage than immediately after harvest, but more intensive odour of a plum jam (Table 2). Flesh firmness, flesh fibrousness and skin hardness were lower after storage (Table 3). Sweet taste was less intensive and bitter taste was stronger after storage (Table 4). Decreasing of sweet taste intensity can results from decreasing sugars comtent in eggplant fruits during storage, what was determined in previous work (Gajewski 2000b). Increasing of bitter taste intensity may results from increasing of polyphenolic acids content in fruits during storage, what was reported in other work (Gajewski & Arasimowicz 2006). In case of fruits of cultivars grown in the greenhouse differences in odour descriptors were found in odour of boiled fungi intensity and odour of boiled vegetables intensity (Table 5). Flesh colour, flesh juiciness, skin hardness, sweet taste, flavour of boiled fungi, bitter taste and sharp taste were also affected by storage (Table 6, 7). In opposite to fruits from the plastic tunnel, storage of fruits from the greenhouse resulted in decreasing skin hardness. In both cases, storage lowered sweet taste intensity and increased bitter taste intensity. Overall quality of fruits of all cultivars was scored on significantly lower level after storage period than immediately after harvest. However, off odour and off-flavour intensity was very low in case of all fruits freshly harvested and stored ones.
Table 2. Results of sensory analysis of eggplant from the foil tunnel odour attributes (scale 0-10) Odour Factor Fresh Stored Bibo Long Black sharp 3.15a 2.99a 3.14a 3.00a of steamed of boiled potatoes fungi 4.94b 3.03a 4.18a 2.72a 4.91b 2.91a 4.21a 2.84a of hay 1,77a 1.91a 1.83a 1.85a of plum jam 1.17a 1.99b 1.28a 1.88b of boiled off-odour vegetables 3.06a 0.01a 2.76a 0.05b 2.96a 0.04a 2.88a 0.03a
Note: mean values for factors which do not differ according to LSD test at P=0.05 are marked with the same letters Table 3. Results of sensory analysis of eggplant fruits from the foil tunnel colour and texture attributes (scale 0-10) Factor Fresh Stored Bibo Long Black Flesh colour 3.17a 4.72b 4.32b 3.53a Seeds development 3.88a 5.16b 5.17b 3.87a Flesh firmness 7.62b 6.73a 7.12a 7.23a Flesh Flesh Skin juiciness fibrousness hardness 4.79a 6.08b 6.23b 4.95b 4.84a 4.94a 4.81a 5.37a 5.69a 4.93a 5.55a 5.48a
Note: see Table 1
Table 4. Results of sensory analysis of eggplant from the foil tunnel flavour attributes and overall quality (scale 0-10) Flavour (taste) of boiled of roasted bitter fungi fruits 1.75a 2.48a 2.18a 1.87a 2.32a 2.51b 1.72a 2.47a 2.09a 1.90a 2.33a 2.59b Overall quality 5.28b 4.88a 5.04a 5.12a
sweet Fresh 2.79b Stored 2.14a Bibo 2.64b Long Black 2.29a Note: see Table 1
sharp. pungent 2.45a 3.40b 2.89a 2.96a
off-flavour 0.05a 0.10b 0.04a 0.11a
Table 5. Results of sensory analysis of eggplant from the greenhouse odour attributes (scale 0-10) Odour sharp Fresh Stored Bibo Long Black 3.34a 3.45a 3.24a 3.55a of steamed of boiled potatoes fungi 5.31a 3.55a 5.20a 3.65b 5.26a 3.86b 5.25a 3.34a of hay 1.71a 2.47b 2.13a 2.05a of plum of boiled off-odour jam vegetables 2.07a 4.00a 0.04a 2.26a 4.12b 0.05a 2.32a 4.33b 0.00a 2.02a 3.80a 0.08a
Note: see Table 1 Table 6. Results of sensory analysis of eggplant from the greenhouse colour and texture attributes (scale 0-10) Flesh colour 1.76a 4.12b 3.59b 2.30a Seeds development 2.84a 3.96a 3.33a 3.45a Flesh firmness 7.27a 7.14a 7.27a 7.14a Flesh juiciness 5.86b 5.54a 5.97b 5.43a Flesh fibrousness 7.15a 6.71a 6.96a 6.91a Skin hardness 4.84a 5.31b 5.26b 4.89a
Fresh Stored Bibo Long Black
Note: see Table 1 Table 7. Results of sensory analysis of eggplant from the greenhouse flavour attributes and overall quality impression (scale 0-10) Flavour (taste) of boiled of roasted bitter fungi fruits 3.21b 3.34a 0.48a 3.03a 3.46a 0.75b 3.33b 3.49a 0.68a 2.91a 3.30a 0.55a Overall quality 6.82b 6.30a 6.70b 6.42a
sweet Fresh Stored Bibo Long Black 3.40b 3.26a 3.45a 3.22a
sharp, off-flavour pungent 1.30a 0.03a 1.84b 0.03a 1.53a 0.00a 1.60a 0.06a
Note: see Table 1
Differences between both cultivars grown in the foil tunnel were significant in sweet taste and bitter taste intensity, and in case of fruits grown in the greenhouse in flesh juiciness and skin hardness. Comparing cultivars from the greenhouse, higher overall quality showed fruits of Bibo, but overall quality of the cultivars grown in the foil tunnel were scored on similar level. In order to find the influence of sensory quality descriptors on quality characteristics of cultivars in relation to term of evaluation, the principal component analysis (PCA) was performed. PCA projections for sensory descriptors and fruit samples are presented on Fig. 1 and 2. The first projection (for fruits from the foil tunnel) shows that two principal components (PC 1 and PC 2) explain together 86% of the variation between samples, with the first component alone accounting for 54.5% of the variation (Fig. 1). The second projection (for fruits from the greenhouse) shows that two principal components (PC 1 and PC 2) explain together 87% of the variation between samples, with the first component alone accounting for 45% of the variation (Fig. 2). The relationship between sensory attributes and fruit samples can be determined by their location on the projection. For the fruits grown in the foil tunnel, points A and C are situated close to each other and in the same direction from the central point of vectors, what indicate similar sensory characteristics of adequate fruit samples freshly harvested fruits of Bibo and Long Black cultivars (Fig. 1). These points are close to the vector of overall quality (vector 20), what indicates better quality of these samples compared with the stored fruits (points B and D). Vector of overall quality has opposite direction compared with vectors of odour of plum jam, of flavour of boiled fungi and of bitter taste, so the influence of these quality traits on the overall quality impression is negative. As it can be seen, fruits of stored both cultivars differed noticeably between each other and from freshly harvested ones. Quite different influence of storage shows PCA for fruits harvested from the greenhouse (Fig. 2). Points B and D (stored fruits of Bibo and Long Black cultivars) are situated on the top of the graph (not on the right side, as in previous projection), and on the perpendicular direction to vector of overall quality. This points out different influence of storage on fruits harvested from the greenhouse, compared to fruits from the foil tunnel. Moreover, vector of overall quality has opposite direction to vectors of sharp odour and pungent flavour intensity (vectors 1 and 18). The reason of different quality changes during storage of fruits from the foil tunnel compared with fruits from the greenhouse can be explained by differences in growing conditions. However, none of these quality changes were so deep to discriminate the fruits. To describe the relationship between scores for overall sensory quality and for sensory descriptors, a linear multiple regression model was applied. In this modeling the assumption was done, that the relationship is of a linear character. It is a simplification however, since there are reports on nonlinear relationship between the intensity of some sensory attributes and overall sensory quality (Meilgaard et al. 1999). The R2 statistics indicates that the model explains 68%
of the variability of overall quality (the relationship is significant at p 0.05), what is quite high value. The equation of the fitted model is as follows: Yo = 2.434 + 0.996x1 + 0.032x2 + 0.040x3 0.126x4 0.107x5 0.061x6 0.482x7 0.052x8 0.104x9 + 0.050x10 + 0.160x11 + 0.131x12 + 0.149x13 + 0.202x14 + 0.129x15 + 0.238x16 0.200x17 + 0.059x18 0.275x19 where: Yo score expected for overall quality of eggplant fruit; x1 - x19 independent variables, expressed by scores obtained for sensory descriptors of fruits and numbered according to Table 1. Overall quality and all descriptors are expressed in numerical values from 0 to 10 units.
5.4
PC2 31.8%
3.4 1 1.4 14 2 20 8 7
-0.6
6 16 C 13 12 3 10
4 18 11 5 17 15 19
D
-2.6 -4.1 -2.1 -0.1 1.9 3.9
PC1 54.5%
Vectors: 1 sharp odour, 2 odour of steamed potatoes, 3 odour of boiled fungi, 4 odour of hay, 5 odour of plum jam, 6 odour of boiled vegetables, 7 off-odour, 8 flesh colour, 9 seeds development, 10 flesh firmness, 11 - flesh juiciness, 12 flesh fibrousness, 13 - skin hardness, 14 sweet taste, 15 flavour of boiled fungi, 16 flavour of roasted fruit, 17 - bitter taste, 18 - pungent flavour, 19 - off-flavour, 20 overall quality. Cultivars / terms: A Bibo fresh, B Bibo after storage, C Long Black fresh, D Long Black after storage. Fig. 1. PCA projection for sensory analysis of eggplant from the foil tunnel. PC1 principal component 1, PC2 principal component 2.
5.2
3.2
B
19 7 1
PC2 42.0%
1.2 9 2
D
18 4 1716 13 8
-0.8
12 20 10 11 14 15
3 6 5
A
-2.8 -3.6 -1.6 0.4 2.4 4.4
PC1 45.2%
Vectors: 1 sharp odour, 2 odour of steamed potatoes, 3 odour of boiled fungi, 4 odour of hay, 5 odour of a plum jam, 6 odour of boiled vegetables, 7 off-odour, 8 flesh colour, 9 seeds development, 10 flesh firmness, 11 - flesh juiciness, 12 flesh fibrousness, 13 - skin hardness, 14 sweet taste, 15 flavour of boiled fungi, 16 flavour of a roasted fruit, 17 - bitter taste, 18 - pungent flavour, 19 - off-flavour, 20 overall quality. Cultivars / terms: A Bibo fresh, B Bibo after storage, C Long Black fresh, D Long Black after storage. Fig. 2. PCA projection for sensory analysis of eggplant from the greenhouse. PC1 principal component 1, PC2 principal component 2.
CONCLUSIONS 1. Three-week storage of eggplant fruits in optimal conditions results in significant changes of their sensory characteristics, what causes decreasing of their overall sensory quality. 2. The main sensory attributes affected are sweet taste, bitter taste and sharp taste intensity. Also flesh colour and intensity of some odour and texture attributes change noticeably. However, the intensity of negative attributes off-odour and off-flavor are still on the low level after storage. 3. Quality changes of fruits from plants grown in the foil tunnel and in the greenhouse differ mainly in case of texture attributes.
4. Overall quality score of eggplant fruits can be predicted with quite high confidency level, using the set of sensory descriptors chosen for the experiment.
REFERENCES Abbott J. 1999. Quality measurement of fruits and vegetables. Postharvest Biol. Technol.: 15: 207-225. Abe K., Ogata K. 1978. Chilling injury in eggplant fruits. J. Jap. Soc. Hort. Sci. 47: 111-120. Anonymous 1996. Sensory analysis. Experts. PN-ISO 8586-2. Anonymous 1998. Sensory analysis. PN-ISO 8589. Anonymous 1999. Sensory analysis. Methodology. PN-ISO 6564. Aubert S., Pochard E. 1981. Problemes de conservation en frais de laubergine (Solanum melongena L.). Rev. Hort. 216: 35-41. [in France] Chabanet C. 2000. Statistical analysis of sensory profiling data. Graphs for presenting results (PCA and ANOVA). Food Qual. Prefer. 11: 159-162. Esteban R., Molla E., Robredo L., Lopez-Andreu F. 1992. Changes in the chemical composition of eggplant fruits during development and ripening. J. Agric. Food Chem. 40(6): 998-1000. Gajewski M., Gajc-Wolska J. 1998. Yielding of eggplant cultivars in foil tunnel and unheated greenhouse. Zesz. Nauk. Akad. Techn.-Roln. w Bydgoszczy, Roln. 42: 65-72 [Polish with English summary]. Gajewski M. 2000a. Quality changes in stored aubergine fruits (Solanum melongena L.) from a plastic tunnel and a greenhouse in relation to the maturity stage and packing method. I. Physical changes. Folia Hort. 14/1: 119-125. Gajewski M. 2000b. Quality changes in stored aubergine fruits (Solanum melongena L.) from a plastic tunnel and a greenhouse in relation to the maturity stage and packing method. II. Chemical changes. Folia Hort. 14/2: 77-83. Gajewski M., Arasimowicz D. 2004. Sensory quality of eggplant fruits (Solanum melongena L.) as affected by cultivar and maturity stage. Polish J. Food Nutr. Sci. Vol. 13/54 No. 3: 249-254. Gajewski M., Arasimowicz D. 2006. The influence of maturity stage and storage on free polyphenolic acids content in fruits of eggplant cultivars (Solanum melongena L.). Folia Hort. Supl. 2006/1: 128-132. Jha S.N., Matsuoka T. 2002. Surface stiffness and density of eggplant during storage. J. Food Eng. 54: 23-26. Kashyap V., Vinod Kumar S., Collonier C., Fusari F., Haicour R., Rotino G.L., Sihachakr D., Rajam M.V. 2003. Biotechnology of eggplant. Sci. Hortic. 97: 1-25. Meilgaard M., Civille G. V., Carr B. T. 1999. Sensory Evaluation Techniques. 3rd ed., CRC Press, Boca Raton London. Mencarelli F., Botondi R., Moraglia D. 1989. Postharvest quality maintenance of new varieties of tomato, pepper and eggplant with small size fruits: preliminary results. Acta Hort. 224: 235-241. Rohde G. 1976. Auberginen und Zucchini. Ernaehrungs Umschau 23: 838-839. [German] Sudheesh S., Presannakumar G., Vijayakumar S., Vijyalakshmi N. 1997. Hypolipidemic effect of flavonoids from Solanum melongena. Plant Foods for Human Nutr. 51: 321-330.
WPYW PRZECHOWYWANIA NA CHARAKTERYSTYK SENSORYCZN OWOCW OBERYNY (SOLANUM MELONGENA L.) UPRAWIANEJ W TUNELU FOLIOWYM I SZKLARNI Streszczenie Celem pracy byo zbadanie wpywu przechowywania owocw oberyny na ich jako sensoryczn. Owoce zebrano w poowie wrzenia, w fazie optymalnej do konsumpcji i przechowywano przez trzy tygodnie w temperaturze 12C i wilgotnoci wzgldnej 95% RH. W dowiadczeniu uyto odmian Bibo i Long Black. Roliny uprawiano w nieogrzewanym tunelu foliowym i w szklarni. Jako sensoryczn okrelano bezporednio po zbiorze i po przechowaniu, z wykorzystaniem metody ilociowej analizy opisowej (QDA). Ocen wykona zesp 12 przeszkolonych ekspertw. Owoce pieczono przez 40 minut w temperaturze 180C, a nastpnie studzono do temperatury pokojowej. W analizie oceniano 19 wyrnikw jakoci sensorycznej oraz jako ogln. Przechowywanie istotnie wpyno na niektre wyrniki jakoci. Owoce z tunelu foliowego wykazyway mniej intensywny zapach gotowanych ziemniakw po przechowywaniu, ale bardziej intensywny zapach demu liwkowego. Jdrno miszu i wknisto miszu byy mniejsze po przechowywaniu, natomiast twardo skrki mniejsza ni bezporednio po zbiorze. Podczas przechowywania intensywno smaku sodkiego zmniejszya si, a intensywno smaku gorzkiego zwikszya u wszystkich owocw. Owoce po przechowywaniu charakteryzoway si ciemniejszym miszem. W przypadku owocw z uprawy szklarniowej przechowywanie wpyno rwnie na intensywno wyrnikw zapachu. Jako oglna u wszystkich odmian bya nisza po przechowywaniu ni bezporednio po zbiorze, jednak zapach obcy i smak obcy byy mao wyczuwalne. Rnice midzy odmianami uprawianymi w tunelu foliowym byy istotne pod wzgldem natenia smaku sodkiego i smaku gorzkiego, natomiast w przypadku odmian uprawianych w szklarni pod wzgldem soczystoci miszu i twardoci skrki.

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THE EFFECT OF STORAGE CONDITIONS ON RED BEETROOT QUALITY

Maria GAWDA Agricultural University Al. 29 Listopada 54, 31-425 Krakw, Poland
Summary Red beetroots, Opolski and Rocket F1 cvs. were kept in a cold store at 0.5 and 2.0C. Two types of packaging were used: traditional plastic boxes and polyethylene film bags. After 6 months of storage beetroots kept in plastic bags had slight weight loss, whereas a decline in dry weight and soluble sugars in their roots was twice bigger than in beetroots kept in boxes. The pigment content in beetroots was decreasing during storage time but betanine revealed a greater resistance to destruction than vulgaxantine. It led to a change of beetroots colours towards more amaranth shades. key words: red beetroot, storage, weight loss, dry weight, soluble sugars, betalains INTRODUCTION Red beetroots (Beta vulgaris var. conditiva) belong among vegetables traditionally cultivated in our part of Europe. This vegetable enjoys a high interest due to its permanent position in Polish cuisine and application for natural food colourants manufacturing (Gasztonyi et al. 2001). Apart from many dietetic values beetroots contain colour compounds from the betalains group with strong antiradical properties (Pedreno & Escribano 2000). From among the identified four betacyanin pigments: betanine, isobetanine, betanidine and isobetanidine, and two betaxantine pigments: vulgaxantin-1 and vulgaxantin-2, betanine constitutes up to 95% of the total amount of betacyanins and vulgaxantine -1 up to 95% of the total betaxantines (Sobkowska et al. 1991). Advantageous anticancer properties are an important reason for consuming red beetroots during wintertime and early spring. The stored raw material is also used for production of betanine, E-162 colourant which is applied without any limitations for food products colouring because it is safe for consumers. Red beetroots belong to vegetables relatively easy to store, however deviation from traditional storing methods in favour of cold storage requires new recommendations concerning packaging which may be successfully used there.
The paper aimed at determining changes in the content of selected components determining red beetroot quality during their long-term storing in a cold storage at diversified temperatures and in different packaging. MATERIALS AND METHODS The experiment was conducted in the years 2003-2004 and 2004-2005 on two red beetroot cultivars with cylindrical root shapes: Opolski (Plantico Gobiew) and Rocket F1 (Bejo Zaden). The beetroots were harvested in the last decade of October. After cutting of leaves and root cleaning the beetroots were brought to the Experimental Cold Store of the Horticulture Faculty, Agricultural University in Krakow (Southern Poland). Medium sized beetroots were chosen for storing (mean weight of Opolski cv. roots was 190g and Rocket cv. roots was 150 g), damaged or diseased one were rejected. Two kinds of packaging were used: plastic boxes, 20 kg of volume lined with black foil and 25 x 40 cm low density polyethylene bags (LDPE), 0.05mm thick, tightly sealed with sealing tape. Two temperatures of storage were assumed 0.5C and 2C. The beetroots were divided into four storage objects: I. boxes, temperature 0.5C, r.h. 94-96% II. bags, temperature 0.5C, III. boxes, temperature 2.0C, r.h. 88-92% IV. bags, temperature 2.0C. Four replications were used: one box was a replication for objects I and III, whereas in objects II and IV the replication was 12 bags. The beetroots were stored for six months since the end of October till the end of April of the following year. Analyses were conducted on the natural loss of mass, dry weight, and soluble sugars by anthrone method (Yemm & Wills 1954) at the beginning of the experiment and then at one month intervals. Moreover the content of betalain pigments (betanine and vulgaxantine) were assessed by Nilsson method (1970). The results were elaborated statistically by ANOVA in a totally randomised design and the differences between means were evaluated on the basis of t-Student test at confidence interval P=0.05. Because of tendencies recurrent in both years of the investigations and for simplification of result presentation means for two years were used in the paper. RESULTS Natural weight loss of the stored red beetroot have been shown in Table 1. The selected cultivars differed by the amount of weight loss: Rocket beetroots lost on average by 14.2% more fresh mass than Opolski cv. beetroots. The method of storage strongly affected transpiration of the stored beetroots. Weight loss on average approximating 20% were registered in beetroots stored in boxes, significantly diversified due to applied temperature and in favour of 0.5C temperature. Application of low density polyethylene film bags significantly limited the weight loss, on average below 0.5% and no differences between objects
M. GAWDA THE EFFECT OF STORAGE CONDITIONS ... 87 _____________________________________________________________________________________________________
with different storage temperatures were detected. While analysing mean loss of beetroot weight in the successive months of storage, it should be said that beetroots packed in plastic bags were loosing their weight only to a slight degree, about 0.1-0.2% monthly. Weight loss in beetroots stored in boxes were growing fastest during the two first months of storage when they reached between 14.6 and 19.6%. During the subsequent period their growth rate was on average 5% per month. By the end of storage beetroots kept in polyethylene film bags revealed minimum weight loss, usually below one percent, whereas in beetroots kept in boxes the loss reached about 30%.
Table 1. Loss of weight of red beets Opolski and Rocket F1 cvs. in different storage conditions (% f.w.) Objects of storage Boxes temp. 0.5C Months of storage Cultivars Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 0 0.00 a 0.00 a 0.00 a 0.00 a 0.00 a 0.00 a 0.00 a 0.00 a 0a Opolski 8.68 a 1 5.13 b 6.56 b 0.10 a 0.09 a 6.60 b 10.09 c 0.10 a 0.12 a 2 15.45 de 16.86 ef 0.36 a 0.30 a 14.62 d 19.64 g 0.24 a 0.21 a 3 18.33 fg 19.77 g 0.51 a 0.42 a 17.80 f 23.78 ij 0.37 a 0.33 a 4 21.89 h 24.35 jk 0.58 a 0.55 a 22.15 hi 27.38 l 0.55 a 0.45 a 5 25.60 k 27.65 lm 0.79 a 0.75 a 27.84 lm 30.84 n 0.86 a 0.67 a 6 27.82 lm 29.40 mn 0.93 a 0.85 a 33.46 o 35.74 p 1.01 a 0.78 a Means for objects of storage 17.06 b
Plastic bags temp. 0.5C
0.44 a
Boxes temp. 2C
19.28 c
Plastic bags temp. 2C Means for months of storage Means for cultivars
0.40 a
3.60 b 8.46 c 10.16 d 12.24 e 14.37 f 16.25 g
Rocket F1 9.91 b
Note: Values marked with the same letter within means and cooperation do not differ significantly
Changes in dry weight content in the stored beetroots were compiled in Table 2. Rocket red beetroots contained less dry weight (on average by 7.1%) than Opolski cv. On average about 7.0% more of this component was registered in the roots of beetroots packed in boxes than kept in plastic bags. The difference between storage temperatures 1.5C was too small for mean dry weight of beetroots kept in boxes to differ significantly. Also mean dry weight of beet-
roots stored in plastic bags at different temperatures was little diversified. The level of this component in beetroots stored in boxes declined definitely during the last (in Opolski cv.) or the two last months of storage (in Rocket cv.), whereas in beetroots kept in plastic bags it started to decrease already in the first months of the experiment. When the storage was completed beetroots kept in boxes contained on average by 8.9% less dry weight than at the beginning of storage, whereas the beetroots kept in bags had 17.3% less.
Table 2. Contents of dry weight in red beets Opolski and Rocket F1 cvs. in different conditions of storage (% f.w.) Objects of storage Boxes temp. 0.5C Months of storage Cultivars Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 0 16.64 v-x 15.44 op 16.64 v-x 15.44 op 16.64 v-x 15.44 op 16.64 v-x 15.44 op 1 16.68 v-x 15.13 lm 15.60 qr 14.92 jk 16.59 vw 14.88 jk 15.99 s 14.29 h 2 16.62 v-x 15.33 no 16.16 t 14.66 i 16.58 v 15.26 mn 16.34 u 15.03 kl 3 16.67 v-x 15.65 r 15.22 mn 14.11 fg 16.74 wx 15.94 s 15.48 o-q 13.34 b 4 16.70 v-x 15.90 s 14.31 h 14.21 gh 16.75 x 16.72 v-x 15.50 p-r 13.80 d 5 16.60 v-x 14.83 j 14.54 i 13.96 ef 16.66 v-x 14.65 i 14.17 gh 12.86 a 6 15.50 p-r 14.07 fg 13.64 c 13.84 de 15.24 mn 13.83 de 13.20 b 13.34 b Means for objects of storage 15.81 c
14.80 b
Boxes temp. 2C
15.85 c
14.67 a
16.04 f 15.51 d 15.70 e 15.39 c 15.49 d 14.78 b 14.08 a Opolski 15.84 b
Rocket F1 14.72 a
Note: see Table 1
Changes in soluble sugars concentrations in stored red beetroots have been presented in Table 3. Opolski cv. beetroots had on average by 7.4% sugars more than Rocket beetroots. On average by 7.7% soluble sugars more was registered in beetroots stored in boxes than in the ones kept in plastic bags. Diversification in the storage temperatures caused a slight, on average 1.8% difference in soluble sugars level only in the beetroots stored in polyethylene film bags in favour of the ones kept at 0.5C. In individual storage objects soluble sugars concentrations in red beet roots were changing in a similar way until March. In the first month of storage a decline in sugar concentrations by about 7.7% was detected,
a 34.9% increase was noted in the second month, and again a 8.0% decrease in the third month. In February soluble sugars concentrations remained on a similar level, and afterwards their levels in beetroots stored in plastic bags decreased on average about 20% during the final two months, whereas in the beetroots stored in boxes only in the last month of storage (on average by about 13%). The analyses conducted in April revealed on average 10.0% higher sugar content in beetroots packed in boxes and by 9% less sugars in beetroots kept in plastic bags in comparison with beetroots analysed at the beginning of storage.
Table 3. Contents of soluble sugars in red beets Opolski and Rocket F1 cvs. in different conditions of storage (% f.w.) Objects of storage Boxes temp. 0.5C Months of storage Cultivars Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 0 7.80 i-k 7.28 fg 7.80 i-k 7.28 fg 7.80 i-k 7.29 fg 7.80 i-k 7.29 fg 1 7.35 fg 6.93 de 6.30 a 7.23 f 7.31 fg 6.55 b 7.96 k-m 6.06 a 2 9.64 t-w 9.75 u-w 9.81 wx 8.20 mn 9.58 t-w 10.03 x 9.62 t-w 8.51 p 3 9.80 v-x 8.11 l-n 8.92 q 8.53 p 9.21 rs 8.14 mn 8.93 q 7.48 gh 4 9.57 t-v 7.76 i-k 8.20 mn 9.02 qr 9.21 rs 7.88 j-l 9.42 st 7.99 k-m 5 9.26 rs 9.52 tu 8.24 no 8.05 l-n 9.22 rs 9.54 tu 7.62 hi 6.99 de 6 8.45 op 7.70 h-j 6.59 bc 7.15 ef 9.08 qr 7.98 k-m 6.86 d 6.81 cd Means for objects of storage 8.49 c
7.95 b
Boxes temp. 2C
8.48 c
7.81 a
7.54 b 6.96 a 9.39 d 8.64 c 8.63 c 8.55 c 7.58 b Opolski 8.47 b
Rocket F1 7.89 a
Note: see Table 1
Changes in betanine content in the roots of stored red beetroots have been shown in Table 4. The compared cultivars differed by this component concentrations, Opolski beetroots contained by 8.7% more betanine than Rocket beetroots. In beetroots kept in boxes on average 12.5% more betanine was assessed than in ones stored in plastic bags. The greatest decline in this pigment content during storage time, on average 35.5% for all objects, occurred in the first month. Betanine level was showing small, although significant diversification between December and February while a subsequent decline, on average 19.1%
was registered in March. At the final stage of storage changes in red pigment concentrations were not big but exceeded the significance threshold. Dynamics of betanine concentration changes in beetroots from individual storage objects revealed tendencies similar to mean value for individual dates described above. On the final date of analyses it was found that betanine content in red beet roots stored in boxes decreased on average by 42.9% and in the roots of those kept in plastic bags by 42.2% of the initial content.
Table 4. Contents of betanine in red beets Opolski and Rocket F1 cvs. in different conditions of storage (mg g f.w.) Objects of Cultivars storage Boxes temp. 0.5C Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Months of storage 0 0.840 A 0.806 z 0.840 A 0.806 z 0.840 A 0.806 z 0.840 A 0.806 z 1 0.675 y 0.504 l-n 0.480 ij 0.406 c 0.563 s 0.597 vw 0.502 lm 0.526 pq 2 0.679 y 0.527 pq 0.474 hi 0.438 f 0.585 uf 0.562 s 0.539 qr 0.488 jk 3 0.685 y 0.619 x 0.468 g-i 0.468 g-i 0.599 w 0.529 pq 0.587 u-w 0.456 g 4 0.548 r 0.510 m-o 0.479 ij 0.422 de 0.414 cd 0.422 de 0.390 b 0.388 b 5 0.571 st 0.510 m-o 0.527 pq 0.464 gh 0.579 tu 0.496 kl 0.432 ef 0.412 cd 6 0.517 n-p 0.358 a 0.422 de 0.500 k-m 0.492 jkl 0.516 n-p 0.518 op 0.462 gh Means for objects of storage 0.596 d
0.513 a
Boxes temp. 2C
0.571 c
0.524 b
0.823 g 0.531 d 0.536 e 0.551 f 0.446 a 0.498 c 0.473 b Opolski 0.574 b
Rocket F1 0.528 a
Note see Table 1
Changes in vulgaxantine concentrations during red beetroot storage have been shown in Table 5. Rocket beetroots had on average by 9.7% vulgaxantine more than Opolski cultivar. On average for the mode of storage beetroots kept in boxes contained by 30.0% more of the yellow pigment than beetroots packed in plastic bags. The effect of temperature on vulgaxantine concentration in red beet roots was small, nevertheless the beetroots kept at 2C has significantly less of this pigment than those stored at 0.5C. A decrease in vulgaxantine level in beetroots during the subsequent months of storage was quite steady and amounted to average of 11.0% to 17.6% during a month. Also in individual
storage objects quite regular declines in this pigment content were noticed, except for Opolski beetroots packed in boxes, in which the level of yellow pigments rose after the first month of storage. Mean content of vulgaxantine in red beet roots decreased by 59.3% of the initial content after six months of storage when they were stored in boxes and by 60.6% if stored in plastic bags.
Table 5. Contents of vulgaxantine in red beets Opolski and Rocket F1 cvs. in different storage conditions (mg g f.w.) Objects of Cultivars storage Boxes temp. 0.5C Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Opolski Rocket F1 Months of storage 0 0.618 r 0.672 t 0.618 r 0.672 t 0.618 r 0.672 t 0.618 r 0.672 t 1 0.675 t 0.624 r 0.459 o 0.624 r 0.669 st 0.723 u 0.348 hij 0.417 mn 2 0.654 s 0.589 q 0.379 k 0.428 n 0.485 p 0.678 t 0.351 h-j 0.393 kl 3 0.624 r 0.432 n 0.300 g 0.285 e-g 0.357 ij 0.615 r 0.348 h-j 0.342 hi 4 0.459 o 0.402 lm 0.290 fg 0.360 j 0.279 d-f 0.411 m 0.288 e-g 0.336 h 5 0.428 n 0.273 de 0.234 c 0.267 d 0.246 c 0.411 m 0.231 c 0.234 c 6 0.279 d-f 0.174 b 0.153 a 0.336 h 0.238 c 0.359 j 0.291 fg 0.240 c Means for objects of storage 0.493 d
0.386 b
Boxes temp. 2C
0.483 c
0.365 a
0.645 g 0.567 f 0.495 e 0.413 d 0.353 c 0.291 b 0.259 a Opolski 0.412 a Rocket F1 0.452 b
Note see table 1
DISCUSSION Assumed temperatures 0.5C and 2.0C considered advantageous for red beetroot storage were so little diversified that their effect on the content of investigated components was similar. The greatest differences in the quality of stored beetroots were caused by different properties of packaging used and especially their different abilities of maintaining high moisture content around the roots. The use of plastic packaging during red beetroot storage most strongly affected the amount of transpiration. Red beet roots have at thin and delicate skin and once take out of the soils are poorly adjusted for restraining water es-
cape. Moreover, beetroots with elongated shapes transpire more easily than these with round roots. It has been demonstrated that by the end of storage weight loss in beetroots stored in LDPE film were 30 times smaller than in beetroots kept in traditional boxes. Therefore plastic packaging allowing to maintain high juiciness, which is one of the most important features of beetroot commercial quality, should be particularly recommended for their storage. However, the use of this type of packaging may cause a problem because of a tendency to resume leaves and roots growth which appears by the end of storage period. Tessarioli and co-workers (1998) suggest a possible application of perforated plastic bags or PVC film, although in that case weight loss are bigger. Due to a considerably higher transpiration, dry weight of beetroots stored in boxes was higher than in beetroots kept in plastic bags. Irrespective of the packaging the beetroot dry weight declined by the end of the experiment, however in beetroots packed in plastic bags the decline was twice bigger than in beetroots placed in boxes. The fact that apparent declines in dry weight appeared only at the final stage of storage when substrate decomposition processes competing with transpiration process started to dominate, should be ascribed to a considerable water loss in beetroots stored in boxes. Similar observations concerning the dynamics of dry weight changes in red beetroots stored traditionally may be found in the paper by Elkner et al. (1997). Beetroots kept in boxes had on average more soluble sugars than beetroots stored in plastic bags and the difference between these components concentrations was proportional to the difference of dry weight of beetroots from these two types of packaging. The analysis of anisotropic changes of soluble sugar levels in beetroots during their storage allows to distinguish the period between the end of November and the end of December when a high increase in their content occurred caused by the decomposition of reserve starch in roots. The other characteristic period is the final stage of storage when sugar content visibly started to decline. However, only in the case of beetroots from plastic bags it was lower at the end than at the beginning of storage. It is considered that red beetroot cultivars with strongly elongated root shape contain less pigment than varieties with round roots (Sobkowska et al. 1991). At the time of harvest Opolski and Rocket c.v. beetroots contained respectively 0.840 and 0.806 mg of betanine per a gram of fresh mass . It is less than in case of beetroots with round root shape, which usually have over 1 mg of this pigment in one gram of root flesh. Beetroots stored in boxes contained more betanine than beetroots packed in plastic bags. Undoubtedly it is the result of clear diversification in dry weight in both storage conditions. The content of betalaine pigments decreases during storage period due to peroxidase activity, which oxidises them to brown polymers with undetermined composition (Pedreno & Escribano 2000). After six months of storage the content of red pigments in beetroots kept in boxes decreased by about 40%. Other authors (Elkner et al. 1997) report similar results which depend on selected varieties and assumed storage temperatures.
Unlike in the case of betanine, Rocket beetroots contained more vulgaxantine. The results of analyses confirmed visual assessment of the colour of selected varieties: Opolski beetroots are characterised by a more amaranthine colour of flesh than Rocket c.v. beetroots. Beetroots stored in boxes had on average more vulgaxantine than those kept in plastic bags and the difference between them was greater in this respect than assessed for betanine. A decrease in yellow pigment content over subsequent months of storage was more regular and unchangeable than the red pigment. Studies on thermostability of red beetroot pigments revealed that the energy of betanine degradation is greater than vulgaxantine, which is more sensitive to destruction factors than betanine (Saguy 1979, Sapers & Hornstein 1979). The betanine to vulgaxantine content ratio after Opolski beetroot harvesting was 1.36 and for Rocket cultivar 1.20. It is below the lower limit of value determined for good quality of beetroot juice which ranges minimum between 1.5 and 1.8 (Czapski & Sobkowska 1990). The ratio changed positively during storage and by the end of this period was on average 2.12 for Opolski beetroots and 1.72 for Rocket cv. The increase in betanine to vulgaxantine content ratio which occurred on all storage objects resulted in a change in beetroot flesh colour towards a more vivid red. The authors researching content of betalaine pigments in red beetroots during cultivation report that the level of betaxantine is increasing constantly until the end of vegetation, which causes that the quantitative growth of these pigments is higher than increase in anisotropically changing betacyanes (Watson & Gabelman 1982, Michalik et al. 1995). In result the betacyanes to betaxantines ratio decrease with delayed harvest causing a change in beetroot colour (Wolyn & Gabelman 1986). Definite declines in vulgaxantine concentrations and less regular decline in betanine content are observed during storage. Therefore the situation is analogous, although the direction of changes is opposite. CONCLUSIONS 1. Red beetroots stored for 6 months in LDPE film bags had about 1% weight loss, 30 times lesser than weight loss of beetroots kept in traditional boxes. 2. A decline in dry weight content of beetroots stored in boxes was twice lower than in beetroots kept in plastic packaging. 3. Soluble sugar concentrations assessed by the end of storage was lower in beetroots kept in plastic bags than in beetroots stored in boxes proportionally to changes of their dry weight. 4. Betanine level in beetroots most strongly decreased in the first month of storage and then in March. Finally the content of red pigment was by about 40% lower by the end of the experiment than at its beginning. 5. Vulgaxantine content in beetroots was steadily declining during storage and after 6 months the decline reached about 60% of this pigment initial content.
6. The ratio of betanine to vulgaxantine content was growing during storage: between 1.36 and 2.12 in Opolski cultivar and between 1.20 and 1.72 in Rocket cv. It caused a change of red beetroot colour from bright red observed by the end of October to amaranthine red by the end of April.
REFERENCES Czapski J., Sobkowska E. 1990. Krajowe preparaty barwnikw czerwonych z buraka wikowego. Przem. Ferment. Owoc. Warz. 11-12: 30-41. [in Polish] Elkner K., Badeek E., Adamicki F. 1997. [The effect of storage conditions on quality of red beet.] Biul. Warzywn. 46: 67-78. [in Polish with English summary] Gasztonyi M.N., Hussein D., Hajos M.T., Biacs P. 2001. Comparison of red beet (Beta vulgaris var. conditiva) varieties on the basis of their pigment components. J. Sci. Food Agric. 81(9): 932-933. Michalik B., ukowska E., lczek S. 1995. Changes in quality of red beet cultivars with growing time. Folia Hort. 7(1): 127-136. Nilsson T. 1970. Studies into the pigments in beetroot. Lantbr. Hgsk. Annlr. 36: 179-219. Pedreno M.A., Escribano J. 2000. Studying the oxidation and the antiradical activity of betalain from beetroot. J. Biolog. Educ. 35(1): 49-51. Saguy J. 1979. Thermostability of red beet pigments (betanine and vulgaxantin-1): influence of pH and temperature. J Food Sci. 44: 1554-1555. Sapers G.M., Hornstein J.S. 1979. Varietal differences in colorant properties and stability of red beet pigments. J. Food Sci. 44: 1245-1248. Sobkowska E., Kaczmarek R., Czapski J., Sobiech B., Sikorski K. 1991. Czynniki wpywajce na jako buraka wikowego jako surowca w przetwrstwie i produkcji barwnikw. Przem. Ferment. Owoc. Warz. 2: 18-21. [in Polish] Tessarioli Neto J., Kluge R.A., Jacomino A.P., Scarpare Filho J.A., Iwata A.Y. 1998. Storage of beetroots Early Wonder in different kinds of package. Horticultura Brasileira. 16(1): 7-10. Watson J.F., Gabelman W.H. 1982. Seasonal changes and cultivar differences in pigment concentrations and percent dissolved solids in roots of table beets. J. Amer. Soc. Hort. Sci. 107(5): 713-716. Wolyn D.J., Gabelman W.H. 1986. Effect of planting and harvest date on betalain concentrations in three table beet genotypes. HortSci. 21(6): 1339-1340. Yemm E.W., Wills A.J. 1954. The estimation of carbohydrates in plant extracts by anthrone. Biochem. J. 57: 508-514. WPYW WARUNKW PRZECHOWYWANIA NA JAKO KORZENI BURAKA WIKOWEGO Streszczenie Buraki wikowe odmian Opolski i Rocket F1 przechowywano w chodni zwykej w temp. 0,5 i 2,0oC. Zastosowano dwa rodzaje opakowa: tradycyjne skrzynki plastikowe i worki z folii polietylenowej. Po 6 miesicach przechowywania buraki przechowywane w opakowaniach foliowych miay znikome ubytki masy, natomiast spadek zawartoci suchej masy i cukrw rozpuszczalnych by dwa razy wikszy ni u burakw przechowywanych w skrzynkach. Zawarto barwnikw burakw obniaa si w czasie przechowywania, przy czym betanina wykazaa wiksz odporno na destrukcj ni wulgaksantyna. Spowodowao to zmian zabarwienia burakw na bardziej amarantowe.

CONTENT OF ISOTHIOCYANATES AND FLAVONOLS IN ROOTS DURING VEGETATION OF TWO TYPES HORSERADISH
Marcin HORBOWICZ1, Maria ROGOWSKA Research Institute of Vegetable Crops Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland
Summary During two-year studies allyl isothiocyanate (AITC), phenetyl isothiocyanate (PEITC) and kaempferol concentration in roots of creamy and white types of horseradish during its vegetation were measured. Content of AITC in horseradish roots ranged from 800 mg to 2000 mgkg-1 FW. In 2002 season similar contents of AITC were found in both types of horseradish, but in 2003 roots of white type contained significantly higher level of that isothiocyanate. Concentration of PEITC in roots of both horseradish types was 3 to 10 times lower than AITC. In almost all terms of analysis roots of creamy type horseradish contained higher level of PEITC than white type. Decline of PEITC level was observed during vegetation of horseradish. Kaempferol level decreased in horseradish roots during cultivation period too, and only traces of quercetin in roots of both horseradish types were noticed. key words: horseradish, Armoracia rusticana, roots, allyl isothiocyanate, phenetyl isothiocyanate, kaempferol INTRODUCTION Epidemiological studies have documented the important role of diets high in cruciferous vegetables in reducing the incidence of human cancers (Stoewsand 1995, Johnson 2002, Horbowicz 2003). Anticarcinogenic properties of the vegetables are result of combination of many substances acting simultaneously. The analysis of data from many cohort studies concluded that high consumption of Brassica vegetables is associated with a reduced risk of cancer of lung, stomach, colon and rectum, and a decrease in total cancer incidence (Morse et al. 1993, Hecht 1996, Johnson 2002). Recently were carried out studies demonstrate a reduced risk for prostate cancer with high consumption of vegetables, particularly cruciferous vegetables (Steinmetz & Potter 1991 ). It is believed that one of the most nutritionally important class of compounds occurred in cruciferous vegetables are glucosinolates, and products of its break_______________________________________ 1
present address: University of Podlasie, Institute of Biology, Department of Plant Physiology and Genetics, Prusa 12, 08-110 Siedlce, Poland
down isothiocyanates (ITCs). ITCs are released from glucosinolates by the action of the enzyme myrosinase (Fahey et al. 2001). Cooking destroys myrosinase, what significantly reduces the ITCs bioavailability from cruciferous vegetables. Phenolic compounds are responsible for the taste and colors of many fruits and vegetables, and protect plants from diseases, ultraviolet light and take part in plant defense system against herbivores as well (Herrmann 1988). One of the most nutritionally important classes of polyphenols is flavonoids. Flavonoids have been shown in a number of studies to be potent antioxidants, capable of scavenging hydroxyl radicals, superoxide anions, and lipid peroxy radicals. Epidemiological studies carried out in last years have indicated that consumption of vegetables rich in flavonoids is associated with reduced risk of cancer and coronary diseases (Hertog et al. 1993, Formica & Regelson 1995, Horbowicz 2000). Glycosides of two flavonols - quercetin and kaempferol are particularly important, due to its wide distribution within the plant kingdom. Vegetables, fruits and beverages are the main dietary sources of flavonols in European diet, primarily as quercetin and kaempferol (Herrmann 1985, Hertog et al. 1992, Horbowicz 2000). Horseradish (Armoracia rusticana) is a perennial plant native to southeastern Europe and western Asia. As a medicinal herb, horseradish strongly stimulates the digestion, increasing gastric secretions and appetite, and promotes perspiration (Ward 1936). Horseradish root contains high level of vitamin C, valuable minerals (potassium, magnesium, calcium, phosphorus, iron) and several glucosinolates. Main glucosinolate in horseradish root is sinigrin which being crushed is enzymatically converted into allyl isothiocyanate (AITC) (Fahey et al. 2001). The allyl isothiocyanate found to be effective in rats as a inducer of phase II enzymes, and it suggests that human regularly consumed of food contained high level of AITC, could contribute to the lower incidence of bladder cancer (Munday 2002). Another glucosinolate of horseradish root gluconasturtiin, decomposes under action of myrosinase into phenetyl isothiocyanate (PEITC) (Fahey et al. 2001). PEITC has been shown to inhibit induction of lung and esophageal cancer in both rat and mouse tumor models (Morse et al. 1993, Hecht 1996, Stoner & Morse 1997). Adesida et al. (1996) demonstrated a pronounced antiproliferative effect of PEITC metabolites on human leukemia cells in vitro, too. There are number of horseradish products commercially available: cream-style prepared horseradish, horseradish contained sauces, beet horseradish and dehydrated horseradish. In Poland a ground horseradish roots are mixed with cooked red beets to obtain traditional food appetizer (called wika) used mainly at Easter time. It is used for sandwiches, and as important taste additive to meat and cooked eggs. Horseradish roots are used as preservative and for quality retaining of fermented and canned cucumbers - very popular components of Poles diet. Additionally horseradish is taken internally against cold and flu in Polish folk medicine, and its leaves are used in compresses against arthritic and rheumatic pains.
M. HORBOWICZ, M. ROGOWSKA CONTENT OF ISOTHIOCYANATES ... 97 _____________________________________________________________________________________________________
Area of horseradish cultivation in Poland reaches 2000 ha, and there are two main types of horseradish grown in Poland: creamy (ca. 80-90% of total production) and white type (10-20%). Earlier studies have shown that plants of creamy type are much more attractive for almost all pest species than white one (Rogowska & Szwejda 2002). Therefore replacing of creamy type horseradish by white one is economically important. There is no information in available literature about isothiocyanates and flavonols in root of mentioned horseradish types and changes during plant vegetation. Thus, the objectives of this study were to compare isothiocyanate and flavonol profiles and its changes during vegetation period in root of two main types of horseradish cultivated in Poland. MATERIALS AND METHODS Horseradish plants were grown in soil (soil type: pseudopodsolic over loamy sand, 1.3% organic matter, pH 6.0) with following fertilizer application: N - 110 kgha-1 (divided into two doses, 60 kg Nha-1 were applied in April, before planting, and 50 kg Nha-1 were applied in June, as topdressing); phosphorus (P2O5 40 kgha-1) and potassium (K2O 120 kgha-1) were applied in Autumn, prior to season of horseradish cultivation. The plants were grown on the experimental field trials of the Research Institute of Vegetable Crops in Skierniewice, in two row replicates (30 plants on 10 m length of row, 67.5 cm spacing between rows) + two out protective rows. From each trial (area of 6.8 m2) 10 horseradish roots were taken to analyses. Roots were analyzed just before planting and during main part of vegetation period: August, September and October. Roots taken to analysis were washed, and one quarter of middle part from each root sample was knife cutting, mixed and taken do glucosinolates and flavonoid determination. From each root sample two laboratory analyses were carried out. Horseradish roots harvested in October 2002 were stored during winter time (November 2002 April 2003) in outdoor conditions covered with sand. Roots infested with pathogens and/or damaged by pests were discarded before planting. Before planting a representative sample of 30 roots was taken to chemical analysis. Standards of allyl isothiocyanate (AITC) was purchased from BDH (UK), phenetyl isothiocyanate (PEITC) from Aldrich-Sigma, and phenyl isothiocyanate (internal standard) from Merck Schuchardt (Germany). Standards of quercetin and kaempferol were purchased from Aldrich-Sigma. Other solvents and reagents of appropriate purity were obtained from POCH, Gliwice (Poland) Isothiocyanates in horseradish tissue were determined according to method described by Sultana et al. (2002) with some modifications. Fresh tissue of horseradish roots (5 g) were homogenized in 200 mL of distilled water. The homogenate was left in room temperature for 2 hours to finish conversion of glucosinolates into appropriate isothiocyanates by endogenous myrosinase. Obtained isothiocyanates were extracted from filtered slurry (5 cm3) by shaking with methylene chloride (2 cm3) contained 0.25 mg of internal standard (phenyl isothiocyanate). To separate organic and water layers samples were centrifuged
for 5 minutes at 3000 g. Lower organic layer were withdrew from centrifuge vial using Pasteur pipette. Isothiocyanates were analyzed on gas chromatograph (Shimadzu GC 17A) equipped in flame ionization detector (FID); chromatographic conditions: column 30 m x 0.32 mm I.D., stationary phase: HP-5 (5% crosslinked methylsilicone layer) 0.25 m layer; column oven temperatures - initial: 40C for 1 min., then increased to 100C, rate 5C min-1; then increased to 240C at rate 10C min-1; and held at 240C for 10 minutes; carrier gas (helium) velocity: 56 cm s-1; total pressure: 190 kPa; injection split ratio: 4:1; detector temperature: 250C; and injector port temperature: 220C. Results of isothiocyanates analysis were calculated on basis earlier prepared standard curves of allyl and phenetyl isothiocyanates. The flavonols content was determined according to method described earlier for onion (Horbowicz 1999, Horbowicz & Kotliska 2000). Before analysis the horseradish samples were dried overnight in oven set at 50C. After pulverizing a 500 mg portions were taken to analyses. Flavonol glycosides were extracted by homogenization of samples in mortar with 60% ethanol-water solution. Extracted glycosides were subjected to hydrolysis in 1.2 N HCl. Hydrolysis conditions were following: temperature 100C, time - 30 min. Obtained flavonol aglycones were extracted by triplicate vigorous shaking with ethyl acetate. Each separated upper layer was withdrew and pooled. To analyses of flavonols a HPLC apparatus equipped with UV detector set at 370 nm was used. The quercetin and kaempferol were isocratically separated on Lichrosorb RP18 (4 x 250 mm, 10 m) column, and a mobile phase was methanol: water mixture (55:45, v/v) contained 0.2% orthophosphoric acid. The flow rate was 0.8 cm3min-1. Results of all analyses were graphically and statistically elaborated using Graph Prism computer program. On all graphs results were presented as means of four replicates standard deviation of means. RESULTS AND DISCUSSION Isothiocyanates content Two main isothiocyanates, released from the glucosinolates sinigrin and 2phenylethylglucosinolate by the naturally occurring enzyme myrosinase, were found in horseradish roots. These were allyl isothiocyanate (AITC), and phenylethyl isothiocyanate (PEITC) (Figs 1 & 2). Concentration of AITC in both types of horseradish, white and creamy, ranged from 800 mg till 2000 mgkg-1 fresh weight (FW). In 2002 season quite similar contents of AITC were found in both types of horseradish, but in 2003 roots of white type horseradish contained significantly higher level that isothiocyanate (Fig. 1). Level of AITC in horseradish roots before its planting in spring (April) was higher then during its field vegetation (August October) in 2002, but lower in 2003. The reason of such situation is unknown, although probably effect of various cultivation and weather conditions in 2001 and 2002 could change the sinigrin level or myrosinase activity (Ciska et al. 2000). Another reason can be various condition of winter storage the horseradish roots, which affected chemical composition of horseradish roots.
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Fig. 1. Contents of allyl isothiocynate (AITC) in roots of two horseradish types before planting (April) and during two years of cultivation (results are means of four replicates standard deviation).
Similar concentration of AITC to our results in horseradish produced in New Zealand was found by Sultana et al. (2003). Kushad et al. (2002) have published data concerning glucosinolates level in 32 accessions and cultivars of horseradish from germplasm collection in Illinois, USA. According to their studies horseradish root contained sinigrin in level ranging from 84.6 to 258 mollg-1 DW. After recalculation on basis approximate dry weight (ca. 30%), it means that horseradish root from the collection should contains from ca. 2400 to 7600 mg of AITC in 1 kg of fresh matter. Such results could be obtained in case 100% conversion of sinigrin into AITC. It seems that such rate of conversion by endogenous myrosinase does not occur.
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Fig. 2. Contents of phenetyl isothiocyanate (PEITC) in roots of two horseradish types before planting (April) and during two years of cultivation (results are means of four replicates standard deviation).
Phenetyl isothiocyanate (PEITC) level was found from 3 to 10 times lower than AITC in roots of both types horseradish (Figs 1 & 2). Its concentration ranged in roots of creamy type horseradish from 75 to 480 mgkg-1 FW, and in roots of white type from 60 to 225 mgkg-1 FW. Mentioned earlier Sultana et al. (2003) have found in horseradish root produced in New Zealand content of PEITC: 185 mgkg-1 FW. In all terms of analysis done in 2002 roots of creamy type horseradish contained higher level of PEITC then white type (Fig. 2). In 2003 roots of horseradish used for planting and analyzed in August PEITC level was higher in creamy type horseradish was higher than in white type as well, but in last terms of analyses (September, October) opposite situation was noticed (Fig. 2). Decline of PEITC level was observed during vegetation (August
October) in roots of both types horseradish in 2002, and in creamy type in 2003 only. Flavonols content In roots of both horseradish types only one flavonol kaempferol, was found in measurable level (Fig. 3). No myricetin, and only traces of quercetin were noted in some samples of roots of creamy and white types horseradish during both vegetation seasons (data not shown). Drying of horseradish tissue carried out in middle conditions (50C, 24 h) prior to extraction and analysis was found to be the appropriate analytical procedure, because preliminary experiments showed that flavonol glycosides are stable during such process (data not shown). Concentration of kaempferol was ranging from 5 to 650 mgkg-1 dry weight (DW) and 15 to 600 mgkg-1 DW for roots of creamy and white type horseradish, respectively. In season 2003 slightly higher concentration was found in roots of white type horseradish, in compare to creamy type (Figure 3). Rapid decline of kaempferol level was noted in roots of horseradish in both seasons of cultivation, 2002 and 2003. Concentration of kaempferol was much higher in roots of both horseradish types in season 2003 than in 2002. Kaempferol, as another phenolics, takes part in protection plants from diseases and plant defense system against herbivores as well (Herrmann 1988). Higher level of kaempferol in 2003 is probably due to various biotic stress conditions (pathogens and herbivores occurrence) in both cultivation seasons, what could biosynthesis and accumulation in horseradish roots of phenolic compounds included kaempferol. Contents of kaempferol in horseradish roots found during present studies were quite similar to results published by Eloesser & Herrmann (1975), but much higher to those published by Bilyk & Sapers (1985), or summarized in US Database for the Flavonoid Content (2003). According to Bilyk & Sapers (1985), horseradish root contained 6 mg kaempferol in 1 kg of fresh weight. After recalculation into dry weight basis it is equal ca. 20 mgkg-1 DW. According to US Database for the Flavonoid Content (2003) horseradish root contained no quercetin, and 1.58 mg100 g-1 FW of kaempferol (after recalculation 52 mgkg-1 DW). The discrepancies can be an effect of sampling technique. In our studies level of kaempferol in horseradish roots was much lower than in 2003. During carrying on experiments in 2002 studies light-brown outer layer of horseradish roots was discarded (it was scrapped using knife). In 2003 roots of horseradish were exactly washed in running water, and after careful drying were analyzed. Probably decline in concentration of kaempferol noted in 2002 was the an effect of removing the outer brown layer (Fig. 3). The accumulation of flavonoids in outer layers of plant tissues is especially high, because it creates a kind of chemical barrier against: UV light, pathogens and herbivores (Herrmann 1988). Results presented here show that content of nutritionally important constituents in root of white type horseradish: allyl isothiocyanate, phenetyl isothiocyanate and kaempferol is generally equal, but in some cases higher than in
root of creamy type. Our studies demonstrate that from nutrition point of view white type horseradish can replace the creamy type, more popular among producers in Poland.
700 600 mgg DM 500 400 300 200 100 0
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April
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Fig. 3. Contents of kaempferol in roots of two horseradish types before planting (April) and during two years of cultivation (results are means of four replicates standard deviation). REFERENCES Adesida A., Edwards L.G., Thornalley P.J. 1996. Inhibition of human leukaemia 60 cell growth by mercapturic acid metabolites of phenylethyl isothiocyanate. Food Chem. Toxic. 34: 385-392. Bilyk A., Sapers G.M. 1985. Distribution of quercetin and kaempferol in lettuce, kale, chive, garlic chive, leek, horseradish, red radish, and red cabbage tissues. J. Agric. Food Chem. 33: 226-228.
Ciska E., Martyniak-Przybyszewska B., Kozowska H. 2000. Content of glucosinolates in Cruciferous vegetables grown in the same site for two years under different climatic conditions. J. Agr. Food Chem. 48: 2862-2867. Eloesser W., Herrmann K. 1975. Flavonole und Flavone der Gemsearten. V. Flavonole und Flavone des Wurzelgemses. Zeits. Lebens.-Unters. Forschung 159: 265-270. Fahey J.W., Zalcmann A.T., Talalay P. 2001. The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. Phytochemistry 56: 5-51. Formica J.V., Regelson W. 1995. Review of the biology of quercetin and related bioflavonoids. Food Chem. Toxic. 33: 1061-1080. Hecht S.S. 1996. Chemoprevention of lung cancer by isothiocyanates. Adv. Exp. Med. and Biol. 401: 1-11. Herrmann K. 1988. [On the occurrence of flavonol and flavone glycosides in vegetables.] Zeits. Lebens.-Unters. Forschung 186: 1-5. [in German with English summary] Hertog M.G.L., Feskens J.M., Hollman P.C.H., Katan M.B., Kromhout D. 1993. Intake of potentially anticarcinogenic flavonoids and their determinants in adults in the Netherlands. Nutrition and Cancer 20: 21-29. Hertog M.G.L., Hollman P.C.H., Katan M.B. 1992. Content of potentially anticarcinogenic flavonoids of 28 vegetables and fruits commonly consumed in the Netherlands. J. Agric. Food Chem. 40: 2379-2383. Horbowicz M. 1999. Changes of flavonols content in onion during the vegetation period and storage. Veget. Crops Res. Bull. 50: 81-91. Horbowicz M. 2003. The occurrence, role and contents of glucosinolates in Brassica vegetables. Veget. Crops Res. Bull. 58: 23-40. Horbowicz M., Kotlinska T. 2000. Level of flavonols in wild and cultivated Allium species. Acta Hortic. 517: 375-380. Johnson I.T. 2002. Glucosinolates in the human diet. Bioavailability and implications for health. Phytoch. Rev. 1: 183-188. Kushad M., Juvik J., Klein B., Wallig E., Jeffery E., Chakravarthula S. 2002. Variation of glucosinolates levels among thirty-two accessions of horseradish cultivars. Abstracts of XXVI Int. Hortic. Congress, 2002, Toronto, Canada, p. 67. Morse M.A., Zu H., Galati A.J., Schmidt C.J., Stoner G.D. 1993. Dose-related inhibition by dietary phenethyl isothiocyanate of esophageal tumorigenesis and DNA methylation induced by N-nitrosomethylbenzylamine in rats. Cancer Letters 72: 103-110. Munday C.M. 2002. Selective induction of phase II enzymes in the urinary bladder of rats by allyl isothiocyanate, a compound derived from Brassica vegetables. Nutrition and Cancer 44: 52-59. Rogowska M., Szwejda J. 2002. Ecological observations of Phyllotreta spp., Pieris brassicae, Pieris rapae and Athalia rosae occurrence on horseradish. Veget. Crops Res. Bull. 57: 95-105. Steinmetz K.A., Potter J.D. 1991. Vegetables, fruit, and cancer. II Mechanisms. Cancer Causes and Control 2: 427- 442. Stoewsand G.S. 1995. Bioactive organosulfur phytochemicals in Brassica oleracea vegetables - a review. Food Chem. Toxic. 33: 537-543. Stoner G.D., Morse M.A. 1997. Isothiocyanates and plant polyphenols as inhibitors of lung and esophageal cancer. Cancer Letters 114: 113-119. Sultana T., Savage G.P, McNeil D.L., Porter N.G., Clark B. 2003.Comparison of flavour compounds in wasabi and horseradish. Food Agric. Environ. 1: 117-121.
Sultana T., Savage G.P., McNeil D.L., Porter N., Martin R.J., Deo B. 2002. Effects of fertilisation on the allyl isothiocyanate profile of the above ground tissues in New Zealand grown wasabi. J. Sci. Food Agric. 82: 1477-1482. USDA Database for the Flavonoid Content of Selected Foods. 2003. http://www.nal.usda.gov/fnic/foodcomp. Ward H. 1936. Herbal Manual. L. N. Fowler & Co. Ltd. London, UK. The Southwest School of Botanical Medicine http://www.swsbm.com, pp. 1-115.
ZAWARTO IZOTIOCYJANIANW I FLAWONOLI W KORZENIACH PODCZAS WEGETACJI DWCH TYPW CHRZANU Streszczenie Dwuletnie badania dotyczyy zmian zawartoci izotiocyjanianu allilu (AITC), izotiocyjanianu fenetylu (PEITC) oraz kemferolu w korzeniach podczas wegetacji dwch typw chrzanu: biaego i kremowego. Zawarto AITC w korzeniach chrzanu wahaa si w zakresie od 800 do 2000 mgkg-1 wieej masy. W sezonie wegetacyjnym 2002 zawartoci AITC w korzeniach obu typw chrzanu byy zblione, natomiast w roku 2003 stwierdzono, e korzenie chrzanu biaego zawieray wysze iloci tego skadnika ni kremowego. Stenie PEITC w korzeniach chrzanu byo od 3 do 10 razy nisze ni AITC. W prawie wszystkich terminach analiz stwierdzono, e korzenie chrzanu kremowego zawieray wysze iloci PEITC, ni korzenie chrzanu biaego. Obserwowano stopniowe obnienie zawartoci PEITC w korzeniach podczas sezonu wegetacyjnego. Poziom kemferolu w korzeniach chrzanu take ulega obnieniu podczas wegetacji. Kwercetyna wystpia w korzeniach obu typw chrzanu w ilociach ladowych.

CONTENT OF FLAVONOIDS IN SOME ALLIUM SPECIES GROWN FOR GREEN BUNCHING

Barbara MYSIAK, Maria TENDAJ Department of Vegetable and Medicinal Plants, Agricultural University St. Leszczyskiego 58, 20-068 Lublin, Poland
Summary The studies conducted in the years 2002-2005 included the analysis of the content of flavonoids (per quercetin) in three Allium species. This concerned common onion (Allium cepa L.), shallot onion (Allium cepa var. ascalonicum L.) and Japanese bunching onion (Allium fistulosum L.). The plants subjected to chemical analysis came from field cultivation and forcing in the conditions of heated and unheated greenhouse. Common onion and shallot were cultivated from sets which were planted in autumn (the middle of October) and in spring (the middle of April). Japanese bunching onions (Allium fistulosum L.) were grown from the seedlings planted in the middle of April and the plants subjected to the analysis of the content of flavonoids came from the harvest in the second year of the cultivation (the third 10-days period of April). Among the three species, independently of the cultivation conditions in the field and forcing in a greenhouse, the most flavonoids were found in shallot plants (on average 754.04 mgkg-1 f.w. from field cultivation and 611.43 mgkg-1 f.w. from forcing). Japanese bunching onions contained definitely less of this element from field cultivation (on average 485.73 mgkg-1 f.w.), but a big quantity in the cultivation from forcing, especially in unheated greenhouse (on average 847.85 mgkg-1 f.w.). In all the species the green shoots contained significantly more flavonoids as compared to the pseudostem. key words: flavonoids (per quercetin), Allium species, green onion bunching, green shoots, pseudostem INTRODUCTION Flavonoids are differentiated group of polyphenolic compounds counted to the secondary metabolites of many plant species. Allium vegetables, particularly common onion and shallot, are extremely abundant in flavonoids, especially quercetin (Horbowicz 1999, 2000, Patil et al. 1995, Sellappan & Akoh 2002). Quercetin is a dominating flavonol (flavonoids sub-class) in vegetables and fruits. The quercetin glycosides cover over 90% of all flavonoids in onion (Patil et al. 1995).
The largest amounts of quercetin is contained in dry outer skin of common onion and shallot, namely in red or brown-colored cultivars (Horbowicz 2000, Lachman et al. 2003). Horbowicz & Kotliska (1998) found that Japanese bunching onion (A. fistulosum) contained much more kaempferol than quercetin. Horbowicz (1999) conducted the experiments referring to changes in quercetin and kaempferol contents during onion vegetation period and its postharvest storage. He found that among flavonols, bulbs contained only quercetin and green shoots quercetin and kaempferol. In the growth season quercetin started accumutation in bulbs when leaves began drying up. Further accumulation of quercetin and increase of its content in bulbs occurred after beginning the process of drying bulbs. The level of quercetin in onion bulbs was relatively high and stable during the last 3-4 months of long-term storage. Due to a great dietetic value of Alliums vegetable harvested at early development stages, evaluation of flavonoids contents in edible parts of young plants is an interesting scientific issue. Therefore, the aim of the research was to evaluate the flavonoids contents in green shoots and pseudostems of three Allium species: onion, shallot and Japanese bunching onion. The influence of cultivation conditions on flavonoids levels in these plants was also determined. MATERIAL AND METHODS The studies conducted in the years 2002-2005 included the analysis of the content of flavonoids (per quercitine) in three Allium species. This concerned common onion (Allium cepa L.), shallot onion (Allium cepa var. ascalonicum L.) and Japanese bunching onion (Allium fistulosum L.). The plants subjected to chemical analysis came from field cultivation and forcing in the conditions of heated ( 16-20oC) and unheated ( 6-12oC) greenhouse. Common onion and shallot were cultivated from sets which were planted in autumn (the middle of October) and in spring (the middle of April). The Japanese bunching onions (Allium fistulosum L.) were grown from the seedlings planted in the middle of April and the plants subjected to the analysis of the content of flavonoids came from the harvest in the second year of the cultivation (the third 10-days period of April). Flavonoids contents were determined in green shoots and pseudostems of studied vegetable species. Plants were harvested at the stage corresponding to the usefulness for bunching. Common onion was harvested when plants were 35-40 cm high and produced 5-7 leaves. Shallot was 25-30 cm high and had 1522 leaves during the harvest. The plants of Japanese bunching onion were higher than onion and shallot during the harvest (40-48 cm of height and 22-48 leaves). These Allium species plants were useful for commercial usage according to the Polish Standard (PN-R-75384). Flavonoids contents were determined by means of Christ-Mullers method described in Polish Pharmacopoeia V (1999). The obtained results were statistically evaluated, using Tukeys test, at the significance level 0.05.
B. MYSIAK, M. TENDAJ CONTENT OF FLAVONOIDS ... 107 _____________________________________________________________________________________________________
RESULTS AND DISCUSSION Flavonoids contents (recalculated onto quercetin) significantly depended on cultivation conditions, edible part of plant and genetic features of the studied onions. Shallot was distinguished by the highest flavonoids contents, regardless the cultivation conditions, i.e. in field, in heated and unheated greenhouse. Mean level of flavonoids in shallot obtained from bulb planting in autumn and harvested in spring (middle or end of May) was 824.46 mgkg-1 f.w. Shallot plants from spring of planting bulbs contained less flavonoids (683.63 mgkg-1 f.w., on average). However, comparing to common onion cultivated from sets at the same time, shallot contained significantly (84.31 mgkg-1f.w., on average) more of this compound (Table 1). Japanese bunching onion cultivated in the field contained significantly less of flavonoids in green shoots (894.43 mgkg1 f.w., on average) and in pseudostem (77.04 mgkg-1f.w., on average) than the same parts at shallot and common onion (Table 1 & 2). Flavonoids contents depended on internal temperature during forced in greenhouse. Plants of all three studied species obtained from forcing in unheated greenhouse contained significantly more flavonoids as compared to those from forcing in heated greenhouse. (Table 2 & 3).
Table 1. Content of flavonoids in common onion and shallot for spring harvest with green shoots (mgkg-1 f.w.)
Planting 2002 2003 time green pseudo- green pseudoof bulbs shoots stem shoots stem autumn 1524.50 164.69 1289.99 225.79 spring 1137.43 168.14 676.19 171.21 mean 1330.96 166.41 983.06 198.50 autumn 1406.91 348.35 1215.21 327.38 spring 1189.82 289.97 1012.91 241.81 mean 1298.37 319.15 1114.06 284.59 autumn 1465.70 256.52 1252.60 276.58 spring 1163.62 229.05 844.52 206.51 mean 1314.66 242.78 1048.56 241.54 species time of planting bulbs part of plants (shoots, pseudostem) Mean green pseudoshoots stem 1407.24 195.24 906.78 169.67 1157.01 182.45 1311.06 337.86 1101.37 265.89 1206.21 301.87 1359.15 266.55 1004.07 217.78 1181.61 242.16
Species
mean 801.24 538.23 669.73 824.46 683.63 754.04 812.85 610.93 711.89 36.875 25.033 25.033
Onion
Shallot
Mean
LSD0.05
Especially large amounts of flavonoids were found in Japanese bunching onion (847.85 mgkg-1 f.w., on average 111.13 mgkg-1 f.w., more as compared to from forcing in heated greenhouse. It proves that plants may produce more flavonoids in worse cultivation conditions such as cold, which may be considered as their defensive response. It would be a confirmation of earlier study presented by Maolepsza & Urbanek (2000), Lachman et al. (1999) as well as Sembratowicz & Czech (2005).
Regardless the cultivation conditions, significantly more flavonoids were found in green shoots than pseudostems for all studied species. Flavonoids content in green shoots of onion cultivated in the field was 1157.01 mgkg-1 f.w., of shallot 1206.21 mgkg-1 f.w., and of Japanese bunching onion 894.43 mg.kg-1 f.w. Contents of flavonoids in pseudostems of these plants were 182.45, 301.87 and 77.04 mgkg-1 f.w., respectively (Table 1 & 3). Similar dependence of flavonoids contents in the same species was observed for plants obtained from forcing in greenhouse. Regardless the temperature during forcing (16-20C in heated, and 6-12C in unheated greenhouse), the green shoots contained significantly more flavonoids than pseudostems. It was 13 times more in common onion, 7 times more in shallot, and 29 times more in Japanese bunching onion (Table 2 & 3).
Table 2. Content of flavonoids in common onion and shallot from forcing in greenhouse conditions (mgkg-1 f.w.)
Place of forcing 2003 2004 2005 Mean green pseudo- green pseudo- green pseudo- green pseudo- mean shoots stem shoots stem shoots stem shoots stem 442.73 78.90 519.95 64.32 292.13
Species
heated 631.70 70.59 485.43 43.47 greenhouse Onion unheated 1034.52 73.45 1432.21 41.29 greenhouse mean 833.11 72.02 958.82 42.38 heated green764.20 311.56 825.06 102.50 house Shallot unheated green1600.53 269.93 1046.89 120.37 house mean 1182.36 290.75 935.97 111.44 heated green697.95 191.07 655.24 72.98 house Mean unheated green1317.52 171.69 1239.55 80.83 house mean 1007.73 181.38 947.39 79.90 species LSD0..05 place of forcing part of plant (green shoots, pseudostem)
1277.25 859.99 951.92
93.39 86.14 48.49
1247.99 883.97 847.06
69.38 658.69 66.85 475.41 154.18 500.62
1202.97 1077.44 697.32
92.81 70.65 63.69
1283.46 161.04 722.25 1065.26 157.61 611.43 683.50 109.25 396.37
1240.11 968.71
93.10 78.39
1265.72 115.21 690.47 974.61 112.23 543.42 21.375 21.375 29.073
B. MYSIAK, M. TENDAJ CONTENT OF FLAVONOIDS ... 109 _____________________________________________________________________________________________________
Table 3. Content of flavonids in Japanese bunching onion after forcing in the greenhouse conditions (mgkg-1 f.w.) Place of cultivation Field Heated greenhouse Unheated greenhouse Mean Part of plant 2003 2004 1069.75 122.50 596.12 1456.65 27.81 742.23 1620.17 18.88 819.52 1382.19 56.39 719.29 2005 747.24 55.91 401.57 1452.64 47.78 750.21 1669.13 42.42 855.77 1289.67 48.70 669.18 Mean 894.43 77.04 485.73 1420.57 52.86 736.72 1643.67 52.04 847.85 1319.55 60.64 690.09 19.289 19.289 21.460
LSD0.05
green shoots 866.31 pseudostem 52.72 mean 459.51 green shoots 1352.43 pseudostem 83.00 mean 717.71 green shoots 1641.72 pseudostem 94.81 mean 868.27 green shoots 1286.82 pseudostem 76.84 mean 681.82 place of cultivation part of plant years
Low level (66.85-182.45 mgkg-1 f.w., in onion; 157.61-301.87 mgkg-1 f.w., in shallot; 60.64 mgkg-1 f.w., in Japanese bunching onion) of flavonoids in pseudostems of studied Allium vegetable species is somehow the confirmation of earlier study performed by Horbowicz (1999), in which common onion in early stage of bulb formation contained much less quercetin and kaempferol than in the end of development when leaves began drying up. It is commonly known that a bulb is produced from thickened leaf sheaths that are not green shoots. Pseudostem of young onion plants is a preliminary stage of later formed organ a bulb. CONCLUSIONS 1. Young plants of common onion, shallot and Japanese bunching onion cultivated for green bunching contained quite large amounts of flavonoids (recalculated onto quercetin). Green shoots contained particularly high levels of flavonoids. 2. Among three studied species, shallot only from field cultivation contained the most flavonoids. Green shoots of Japanese bunching onion forced in greenhouse was the most abundant in flavonoids. 3. Regardless the cultivation conditions, pseudostems of all studied species contained much less flavonoids than green shoots. The highest differences occurred in Japanese bunching onion, the lowest in shallot.
Acknowledgement The studies were party sponsored by The Ministry of Science and Higher Education (MNiSzW) within a research project No 2 P06R 015 27.
REFERENCES Horbowicz M., Kotliska T. 1998. [Diversity of flavonol contents in some of wild and cultivated Allium species]. Zesz. Probl. Post. Nauk Roln. 463: 529-537. [in Polish with English summary] Horbowicz M. 1999. Changes of the flavonols content in onion during the vegetation period and storage. Veg. Crops Res. Bull. 50: 81-91. Horbowicz M. 2000. [Occurrence, biosynthesis and biological properties of the flavonols]. Postpy Nauk Roln. 2: 3-18. [in Polish with English summary] Farmakopea Polska [Polish Pharmacopoeia] V. 1999. t. V. Wyd. PTFarm. Warszawa [in Polish] Lachman J., Orsk M., Pivec V. 1999. Flavonoid antioxidants and ascorbic acid in onion (Allium cepa L.) Zahradnictvi (Hort. Sci.) 26 (4): 125-134. Lachman J., Pronk D. Hejtmnkov A., Dudjak J., Pivec V., Faitov K. 2003. Total poliphenol and main flavonoid antioxidants in different onion (Allium cepa L.) varieties. Hort. Sci. (Praque), 30 (4): 142-147. Maolepsza U., Urbanek H. 2000. [Plant flavonoids as biochemical active compounds]. Wiad. Bot. 44 (3/4): 27-37. [in Polish with English summary] Patil B.S., Pike L.M., Yoo KilSun. 1995. Variation in the quercetin content in different colored onions (Allium cepa L.). J. Amer. Soc. Hort. Sci. 120 (6): 909-913. Sellappan S., Akoh C.C. 2002. Flavonoids and antioxidant capacity of Georgia-grown Vidalia onions. J. Agric. Food Chem., 50: 5338-5342. Sembratowicz I., Czech H. 2005. [Natural antioxidants in the food]. Post. Nauk Rol. 1: 75-88. [in Polish with English summary]
ZAWARTO FLAWONOIDW W ROLINACH NIEKTRYCH GATUNKW WARZYW CEBULOWYCH UPRAWIANYCH NA ZBIR PCZKOWY Streszczenie Badania przeprowadzone w latach 2002-2005 obejmoway ocen zawartoci flawonoidw (w przeliczeniu na kwercetyn) u trzech gatunkw warzyw cebulowych cebuli zwyczajnej, szalotki i siedmiolatki, uprawianych na zbir pczkowy. Roliny poddane analizie chemicznej pochodziy z uprawy polowej i pdzenia w szklarni ogrzewanej i nieogrzewanej. Cebul zwyczajn i szalotk uprawiano z dymki (rednica 20-25 cm), ktr sadzono jesieni w poowie padziernika i wiosn w poowie kwietnia. Siedmiolatk uprawiano z rozsady sadzonej w pole w poowie kwietnia, a roliny poddane analizie zawartoci flawonoidw pochodziy z wiosennego zbioru w drugim roku wegetacji (trzecia dekada kwietnia) oraz pdzenia od poowy marca do koca kwietnia. Spord trzech gatunkw, niezalenie od warunkw uprawy w polu oraz pdzenia w szklarni, najwicej flawonoidw zawieray roliny szalotki (rednio 754,04 mgkg-1 w.m. z uprawy w polu i 611,43 mgkg-1 w.m. z pdzenia w szklarni). Roliny siedmiolatki zawieray zdecydowanie mniej tego skadnika z uprawy polowej (rednio 485,73 mgkg-1 w.m.), lecz bardzo duo z pdzenia, zwaszcza w szklarni nieogrzewanej (rednio 847,85 mgkg-1 w.m.). U wszystkich gatunkw szczypior zawiera istotnie wicej flawonoidw ni odyga rzekoma.

EVALUATION OF SOME MORPHOLOGICAL CHARACTERISTICS OF GARLIC (ALLIUM SATIVUM L.) GENOTYPES

Magdalna VALKOV, Juliana KRLOV Research Institute of Vegetables Andovsk 6, 94001Nov Zmky, Slovak Republic e-mail: valsikovam@vuznz.sk
Summary During the years 2003-2005 at Research Institute of Vegetables in Nov Zmky, field variety trials of garlic (Allium sativum L.) were conducted. The aim of the experiments was to observe 14 genotypes of garlic. Evaluation of garlic biological material was focused on selected morphological parameters of plants. Characteristics of garlic leaves, bulbs, cloves and presence or lack of seed stalk were examined. In foliage attributes two genotypes (Povaie 075 and Slokys 308) are very close to registered variety Valko and four other genotypes (Slofat 78, Slofat 102, Slospi 79, Slospi 123) are similar to registered variety Novozmock. According to shape characteristics of garlic bulb the same shape was identified in certified variety Valko and samples of Poana 037, Slofat 102 and Ukrkar 229. The registered variety Novozmock has comparable characteristic of bulb shape to genotypes of Povaie 075, Slogem 302, Slokys 18, Slokys 317 and Slospi 79. Registered varieties Valko and Novozmock were compared with other genotypes according to colour of clove skin and presence or lack of seed stalk. The relationships in these attributes are evident between Novozmock, Slokys 18, Slospi 123 and between Valko, Povaie 075 and Slokys 308. The observed characteristics are important indicators for diversity quality of genetic resource samples. The results occurrence for documentation of plant descriptors and for utilisation in breeding works. key words: garlic, variety trials, morphological parameters, evaluation INTRODUCTION Garlic can be mentioned among the oldest utilised plant species with very favourable nutritional value for human health. Garlic is a vegetable and at the same time a spice with strong aroma. It is widely used in household cooking, in industry for food processing and in pharmacy industry as well. Thanks to breeders there are some domestic and foreign garlic varieties in cultivation. They mostly fulfil requirements of producers, consumers and other users. In the List of registered varieties for year 2005 there are 12 cultivars of
VEGETABLE CROPS RESEARCH BULLETIN 65 112 ________________________________________________________________________________________________
Allium sativum L. recorded. From this number, 11 are winter cultivars and 1 is spring variety. There is necessity to develop new cultivars with additional values. The important step for garlic breeding is continual testing the genetic resources attributes. Part of this work is study and description of garlic collection of various origins. In the past, garlic variety trials were conducted at various localisations by Paszko, Lun (1989), Vitekov (1998), Krlov (2004) and others. Presently tested varieties include regional samples from collecting expedition in the various regions of Slovakia and Ukraine. This collection is compared with domestic cultivars of Slovak origin, which were bred at the Research Institute of Vegetables in Nov Zmky. MATERIALS AND METHODS Eleven germplasm lines of garlic Allium sativum (Poana 037, Povaie 075, Slofat 102, Slofat 78, Slogem 302, Slokys 18, Slokys 308, Slokys 317, Slokys 9, Slospi 123, Slospi 79) came from collecting expedition in the regions of Slovakia and one sample (Ukrkar 229) from Ukraine. Other two genotypes (Novozmock and Valko) are regular commercial domestic cultivars. All samples were studied according to morphology characteristics of plants and bulbs. Variety trials were carried out according to common agronomic technologies (Valkov et al. 1996, Demo & Hriovsk et al. 2002) in the years 2003-2005 under field conditions on sandy soil. Experiments were conducted at the Research Institute of Vegetables in Nov Zmky. Region of Nov Zmky is situated in South Slovakia, 120 m above sea level. The average rainfall is 546 mm per year and the average temperature is 9.9C per year. Selected morphological characteristics were evaluated according to Descriptor for Allium ssp. (Astley et al. 2003). The evaluated attributes represented average values acquired by analyses of ten plants or bulbs in three replications. Quantitative parameters were evaluated by measuring the length and width of leaves. Cloves were weighed on Sartorius 1264 MP digital scales. Other characteristics were evaluated visually (Vitekov 2002). The tested characteristics were foliage density, length, width and colour of leaves and foliage attitude. The bulbs of garlic were described by shape of the compound bulb in horizontal and longitudinal section. The cloves were evaluated by skin colour, weight and number of cloves. The seed stalk was evaluated and recorded as absent or present. RESULTS AND DISCUSSION Characteristics of cultivars foliage are documented in Table 1 (Astley et al. 2003). Foliage density was low in samples Slokys 18, Slokys 317 and Ukrkar 229, and high in Povaie 075, Slokys 308 and Valko. Other garlic germplasms had intermediate density of foliage. Prostrate foliage attitude was detected in three genotypes, Poana 037, Slogem 302 and Slokys 317 and inter-
M. VALSIKOVA, J. KRALOVA EVALUATION OF SOME MORPHOLOGICAL ... 113 _____________________________________________________________________________________________________
mediate in others. The average length of leaves ranged from 320 mm (Slogem 302) to 380 mm (Novozmock) and width of foliage from 27 mm (Slopsi 123) to 34 mm (Novozmock and Slopsi 79). Data about foliage colour indicates green leaves for varieties of Povaie 075, Slokys 308, Slokys 317, Ukrkar 229, Valko, dark green for Slokys 9, Slokys 18 and light green for the other of varieties. According to foliage attributes two genotypes (Povaie 075 and Slokys 308) are very close to registered variety Valko and four genotypes (Slofat 78, Slofat 102, Slospi 79, Slospi 123) are similar to registered variety Novozmock.
Table 1. Characteristics of garlic foliage Leaves Foliage density Novozmock Poana 037 Povaie 075 Slofat 78 Slofat 102 Slogem 302 Slokys 9 Slokys 18 Slokys 308 Slokys 317 Slospi 123 Slospi 79 Ukrkar 229 Valko intermediate intermediate dense intermediate intermediate intermediate intermediate sparse dense sparse intermediate intermediate sparse dense Length (mm) 380 362 367 358 347 320 364 376 342 378 350 379 362 366 Width (mm) 34 31 33 30 29 28 33 32 28 32 27 34 31 30 Colour light green light green green light green light green light green dark green dark green green green light green light green green green Leaf attitude intermediate prostrate intermediate intermediate intermediate prostrate intermediate intermediate intermediate prostrate intermediate intermediate intermediate intermediate
Garlic bulb shape of the compound bulb in horizontal and longitudinal section was described in Table 2. by Astley et al. (2003). By shape of bulbs in horizontal cut the cultivars are divided into two groups. Samples of Slofat 78, Slokys 9, Slokys 308 and Slospi 123 belong to group of bulbs with elliptic shape. All other varieties fall into a group of circular bulbs. By shape of bulbs in longitudinal section, half of varieties had elliptic and the other half wide elliptic shape. By characteristics of garlic bulb shape, the same shape was identified in certified variety Valko and samples of Poana 037, Slofat 102 and Ukrkar 229. The registered variety of Novozmock has similar bulb shape with genotypes Povaie 075, Slogem 302, Slokys 18, Slokys 317 and Slospi 79.
Table 2. Characteristics of garlic bulbs Cultivar Novozmock Poana 037 Povaie 075 Slofat 78 Slofat 102 Slogem 302 Slokys 9 Slokys 18 Slokys 308 Slokys 317 Slospi 79 Slospi 123 Ukrkar 229 Valko Bulb shape in horizontal section circular circular circular eliptic circular circular eliptic circular eliptic circular circular eliptic circular circular Bulb shape in longitudinal section flattened eliptic eliptic flattened eliptic eliptic eliptic flattened eliptic eliptic flattened eliptic eliptic flattened eliptic flattened eliptic flattened eliptic eliptic eliptic
In Table 3 attributes of garlic cloves are presented (Astley et al. 2003).

Table 3. Characteristics of garlic cloves and presence of a seed stalk Cloves Weight (g) 5.0 6.6 4.9 2.6 5.3 2.7 4.8 5.4 5.7 2.8 5.1 4.3 4.6 4.1
Cultivar Novozmock Poana 037 Povaie 075 Slofat 78 Slofat 102 Slogem 302 Slokys 9 Slokys 18 Slokys 308 Slokys 317 Slospi 79 Slospi 123 Ukrkar 229 Valko
Colour of clove skin cream white cream white light violet white light violet cream cream white white cream light violet cream
Number of cloves 11 7 9 12 6 7 7 10 8 8 9 11 7 8
Seed stalk absent present present absent present present present absent present present present absent present present
Skin colour of the cloves was light violet in samples Slofat 102, Slokys 9 and Ukrkar 229. Varieties Novozmock, Valko, Povaie 075, Slokys 18, Slokys 308, Slospi 123 had cream colour of clove skin and white colour was detected in the rest of genotypes. The average number of cloves per compound bulb was from 6 to 12. There were more than 10 cloves in the bulbs of cultivars Novozmock, Slofat 78, Slokys 18 and Slospi 123. The other cultivars had less than 10 cloves in a bulb. The average weight of 10 cloves was between 2.6 and
M. VALSIKOVA, J. KRALOVA EVALUATION OF SOME MORPHOLOGICAL ... 115 _____________________________________________________________________________________________________
6.6 g. The lowest weight (from 2.6 g to 2.8 g) was detected in samples Slofat 78, Slogem 302 and Slokys 317. Medium weight of cloves (4.1-4.9 g) has been identified in cultivars Povaie 075, Slokys 9, Slospi 123, Ukrkar 229 and Valko. The average cloves weight above 4.9 g was measured for varieties Novozmock, Poana 037, Slofat 102, Slokys 18, Slokys 308 and Slospi 79. The seed stalk was present in all genetic resources except for Novozmock, Slofat 78, Slokys 18, and Slospi 123. The registered varieties Novozmock and Valko were compared with other genotypes by colour of clove skin and by presence or lack of seed stalk. The relationships are evident between Novozmock, Slokys 18, Slospi 123 and between Valko, Povaie 075 and Slokys 308. Results of this study indicate significant variation of morphological traits of Allium sativum genotypes. The similar experiments were carried out by Paszko, Lun (1989) with new garlic varieties. Valkov (1985) conducted variety trials of vegetables to evaluate them in large-scale production. Other works evaluated vegetable cultivars from various perspectives (Valkov et al. 2004, Pokluda 2003) and to deliver results to the gene bank. CONCLUSIONS Results from field experiments were obtained from region of south Slovakia. There were investigated genetic resources of Allium sativum. In presented variety trials the medium density of foliage was identified in 8 cultivars. Foliage attitude was prostrate in three genetic resources, Poana 037, Slogem 302 and Slokys 317 and intermediate in others. The average length of leaves ranged from 320 mm (Slogem 302) to 380 mm (Novozmock) and width of foliage from 27 mm (Slopsi 123) to 34 mm (Novozmock and Slopsi 79). Data about foliage colour enable to segregate genotypes as green, dark green and light green. According to shape of bulbs in horizontal cut, samples of Slofat 78, Slokys 9, Slokys 308 and Slospi 123 belong to a group of elliptic bulbs. Skin colour of the clove was light violet in samples Slofat 102, Slokys 9 and Ukrkar 229. The average number of cloves per compound bulb was from 6 to 12. The average weight of 10 cloves was between 2.6 and 6.6 g. The seed stalk was absent in genotypes of Novozmock, Slofat 78, Slokys 18, and Slospi 123. Study of various characteristics of garlic cultivars enables to select recommended biological material for alternative growing technology. Obtained data can be used for breeding purposes.
REFERENCES Astley D. et al. 2003. Descriptors for Allium spp. European Cooperative Programme for Crop Genetic Resources Network. IPGRI Rome: 42. Demo M., Hriovsk I. et al. 2002. Trvalo udraten technolgie v zhradnctve. Vysokokolsk uebnica. SPU v Nitre, FZKI, Katedra trvalo udratenho rozvoja: 581. [in Slovakian]
Krlov J. 2004. Vybran znaky genetickch zdrojov cesnaku, fazule a rekovky v roku 2003. Informan spravodajca Genofond, VRV Pieany 8: 38-39. [in Slovakian] Paszko J., Lun J.1989. [Evaluation of Allium sativum new varieties from the Krov pri Senci crop improvement station from the aspects of fertility, storability and respiration intensity.] Vedeck prce Vskumnho a achtiteskho stavu zeleniny a pecilnych plodn v Hurbanove, Prroda Bratislava 6: 109-117. [in Slovakian with English summary] Pokluda R. 2003. Comparison of selected characteristics of root parsley cultivars. Prague. Hort. Sci. 2: 67-72. Valkov M. 1985. Vber odrd plodovej zeleniny pri vekovrobnej technolgii. Zveren vskumn sprva. Vskumn a achtitesk stav zelenn a pecilnych plodn Hurbanovo: 155. [in Slovakian] Valkov M. et al. 1996. Produkn systmy vybranch druhov zelenn I. as. Vskumn stav zelenn a pecilnych plodn Nov Zmky: 161. [in Slovakian] Valkov M., Krlov J., Belko I. 2004. Evaluation of vegetable pepper assortment. Horticultural Science Zahradnictv Prague 31 (2): 58-62. Vitekov A. et al. 1998. Ochrana genofondu kultrnych rastln v Slovenskej republike. Zveren sprva lohy PO 95/5145/016-08-01. Vskumn stav zeleninrsky Nov Zmky: 52. [in Slovakian] Vitekov A. 2002. Monitoring of genetic resource collections of vegetables and medicinal plants in the year 2000. Scientific and Research Papers, Research Institute of Vegetables Nov Zmky 11: 120-125.
OCENA WYBRANYCH CECH MORFOLOGICZNYCH GENOTYPW CZOSNKU (ALLIUM SATIVUM L.) Streszczenie W latach 2003-2005 w Research Institute of Vegetables w Nov Zmky w dowiadczeniu polowym porwnywano odmiany czosnku (Allium sativum L.). Celem bada bya ocena 14 genotypw czosnku. Ocena materiau biologicznego czosnku obejmowaa wybrane cechy morfologiczne rolin poprzez opis lici, zbkw i gwek oraz obecno lub brak pdu nasiennego. Pod wzgldem cech lici dwa genotypy (Povaie 075 i Slokys 308) s podobne do zarejestrowanej odmiany Valko, a cztery inne genotypy (Slofat 78, Slofat 102, Slospi 79, Slospi 123) s podobne do odmiany Novozmock. Taki sam ksztat gwki stwierdzono u odmiany Valko i linii Poana 037, Slofat 102 oraz Ukrkar 229. Odmiana Novozmock miaa ksztat gwki podobny do genotypw Povaie 075, Slogem 302, Slokys 18, Slokys 317 i Slospi 79. Odmiany Valko i Novozmock porwnywano z badanymi genotypami rwnie pod wzgldem barwy osonki zbka oraz obecnoci pdu nasiennego. Pod wzgldem tych cech stwierdzono wyrany zwizek pomidzy Novozmock, Slokys 18 i Slospi 123 oraz Valko, Povaie 075 i Slokys 308. Badane cechy s wanymi wskanikami zmiennoci genetycznej pomidzy badanymi prbkami.

DISTRIBUTION OF SULFUR-CONTAINING AMINO ACIDS IN FIFTEEN GARLIC VARIETIES

Jana KRLOV1, Jan VELEK1, Jaroslava OVESN2, Helena STAVLKOV2 1 Institute of Chemical Technology, Technick 5, 166 28 Prague 6 Dejvice, Czech Republic e-mail: jana.kralova@vscht.cz 2 Research Institute of Crop Production, Drnovsk 507, 161 06 Prague 6 Ruzyn, Czech Republic
Summary The content of S-allyl-L-cysteine (deoxyalliin) and S-alk(en)yl-L-cysteine sulfoxides (particularly alliin, methiin, and isoalliin), important nonvolatile flavor precursors, was determined in three parts (bulbs, stems, and leaves) of 15 different species of garlic (Allium sativum L.) plants. The analytical method was based on a pre-column derivatization of the individual S-containing amino acids with o-phthaldialdehyde/2-methylpropane-2-thiol. The derivatised amino acids were determined using a reversed-phase high-performance liquid chromatography with UV detection. Statistical evaluation of the achieved results was done employing linear discriminant analysis that enabled characterization and differentiation of the analysed samples. key words: Allium, garlic, flavor precursor, alliin, isoalliin, methiin, deoxyalliin INTRODUCTION Garlic is a member of the genus Allium in the family Alliaceae. Because of its characteristic pungent flavor, garlic has been cultivated since antiquity as a vegetable and flavoring agent. Garlic also has many medicinal properties. Garlic acts as an antimicrobial agent and improves blood sugar metabolism. It helps prevent atherosclerosis and coronary heart disease by reducing platelet aggregation, promoting fibrinolysis, and lowering blood triglyceride and cholesterol levels. The compounds responsible for the characteristic flavor and physiological activities of garlic are primarily organosulfur compounds, many of which have been identified. Among them are sulfur-containing amino acids, including Salk(el)yl-L-cystein sulfoxides (ACSOs), a unique class of sulfur-containing amino acids present in intact garlic bulbs (Kubec et al. 2000, Ichikawa et al. 2006, Velek et al. 2006). When the garlic tissue is crushed or cut, ACSOs are converted into their corresponding thiosulfinates by a C-S lyase called alliinase (EC 4.4.1.4). For ex-
ample, the ACSO alliin is converted into allicin, which is responsible for characteristic flavor and antimicrobial properties of fresh garlic. Among the minor products of the conversion process are dimethyl thiosulfinate and allylmethyl thiosufinate derived from alliin analogs (Velek et al. 1997, Hughes et al. 2005). The genus Allium includes many different species and varieties of garlic. The species most commonly cultivated is Allium sativum. After thousands of years of selection and cultivation, A. sativum has lost the ability to produce fertile seeds. Some varieties have even lost the ability to form flower stalks and flowers. This is presumably due to random mutations which have been selected over the years (University of Minnesota 1999). There are three main groups of cultivated garlic: flowering plants, scape absent and semi-bolters. This classification is based on IPGRI (IPGRI 2001; Research Institute of Crop Production 2005) descriptor one of them the bolting type is. Molecular markers clearly divided characterized genotypes into two groups hardneck and softneck including semi-bolters. Flowering plants varieties produce a flower stalk, and are therefore also known as bolting or hardneck varieties. They are presumably closely related to wild garlic species. The hard flower stalk makes braiding difficult. Another drawback of flowering plants varieties is that they cannot be stored for longer periods because they form roots and start to dry out within a few months of harvest. Semi-bolters and scape absent varieties are subgroups of Allium sativum (L.), var. sativum. Scape absent varieties do not produce a flower stalk at all. Semi-bolters varieties produce flower stalks, but not flowers. However, flower stalk formation however can depend on climatic conditions. It is not underlaid by genetic changes and probably it is connected with gene expression or epigenetic changes. Scape absent varieties never produce flower stalks at all. Semibolters and scape absent varieties exhibit a high degree of geographical diversity (University of Minnesota 1999). They also can be stored for six to eight months without appreciable deterioration, much longer than the flowering plants varieties can be stored. Several methods have been developed to determine ACSO levels. Direct methods measure the levels of the ACSOs themselves. Indirect methods measure the by-products of enzymatic transformation of the ACSOs, such as pyruvic acid, disulfides, vinyldithiins, and ammonia. Direct methods are usually based on either high-performance liquid chromatography (HPLC) or gas chromatography (Velek et al. 1993, Yoo et al. 1998, Kubec et al. 2000). The aim of this study was to determine the levels of four sulfur-containing amino acids in the bulbs, stems and leaves of fifteen varieties of cultivated garlic, and how these levels differ in flowering plants, semi-bolters and scape absent varieties. Three of these amino acids, alliin, methiin and isoalliin, are ACSOs. The fourth amino acid was deoxyalliin, or S-allylcysteine. The target analytes were extracted with methanol and quantified by reversed-phase HPLC.
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MATERIALS AND METHODS The distribution of sulfur-containing amino acids was determined in five flowering plants, five semi-bolters and five scape absent garlic varieties, representing the garlic collection of genebank in the Czech Republic. Live garlic plants were kindly provided by the Research Institute of Crop Production in the Czech Republic. Plants of each variety were divided into three portions: bulbs, stems and leaves. Each portion was immediately analysed for alliin, methiin, isoalliin and deoxyalliin content. S-alk(en)yl-L-cysteines were synthesized by alkylizing L-cysteine with the appropriate alk(en)yl halides. ACSOs were then synthesized by oxidizing the Salk(en)yl-L-cysteines with hydrogen peroxide (Kubec et al. 2000). Phosphate buffers were made by dissolving 7.8 g of dihydrogen phosphate in 1000 ml of distilled water and adjusting to either pH 6.5 or pH 9.5 with sodium hydroxide. The derivatization agent was prepared by dissolving 140 mg of ophthaldialdehyde in 5 ml of methanol, and then adding 0.1 ml of 2-methylpropane2-thiol and 50 ml of pH 9.5 phosphate buffer (Velek et al. 1993). The target amino acids were extracted from 5 grams of fresh plant material by 50 ml of boiling methanol for 10 minutes. This also ensured that the enzyme alliinase was inactivated. Then mg of norleucine was added to the extraction mixture as an internal standard. The mixture was then homogenized in a blender and filtered through a 0.45 m cellulose acetate HPLC syringe-tip filter (Alltech). The filtrate (100 l) was then derivatised with 900 l of the derivatization agent (Velek et al. 1993) and its aliquot was analysed by HPLC. Twenty l samples of the filtered extracts were injected onto a Hypersil column 4.5 mm in diameter and 250 mm long, with an average pore size of 5 m (ThermoQuest, Hypersil Division, UK). The column was perfused with the help of a ConstaMetric pump system (Watrex), with a flow rate of 0.8 ml/min. The eluent was collected with the help of a AS 100 autosampler fitted with a SpectroMonitor 337 nm UV detection system (Thermo Instrument Systems, Inc., USA). Separation was carried out using gradient HPLC with a two-component mobile phase. Component A was prepared by diluting pH 6.5 phosphate buffer (see Materials and Methods) with nine volumes of distilled water. Component B was pure methanol. At the start of the separation process, the proportion of methanol in the mobile phase was 44%. The proportion of methanol was then increased linearly to 75% over 37 minutes. The proportion of methanol was kept at 75% for a further two minutes, after which it was returned to 44%. The data obtained were indexed by plant part and by variety group, and elaborated using linear discriminant analysis (SPSS for Windows, Release 11.0.0, SPSS Inc., USA).
RESULTS AND DISCUSSION Concentrations of sulfur-containing amino acids in different parts of the garlic plant The concentrations of methiin, alliin, isoalliin and deoxyalliin in the bulb, stem and leaf portions of the garlic plants are presented in Tables 1, 2, 3 and 4. The combined concentration of all four amino acids was far higher in the bulb portion than in either the stem or leaf portions. The main sulfur-containing amino acid present in all three portions was alliin, with concentrations far higher than the other three amino acids.
Table 1. Basic statistical data for all samples (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.067 0.771 0.015 0 Maximum 0.802 3.666 0.251 0.206 Average 0.279 1.658 0.066 0.077 2.080 RSD 0.162 0.632 0.047 0.052
Table 2. Basic statistical data for bulbs (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.067 1.473 0.052 0.038 Maximum 0.430 3.666 0.251 0.206 Average 0.204 2.204 0.109 0.103 2.619 RSD 0.118 0.644 0.052 0.052
Table 3. Basic statistical data for stems (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.105 0.771 0.026 0.050 Maximum 0.636 2.445 0.130 0.171 Average 0.260 1.366 0.050 0.100 1.776 RSD 0.149 0.463 0.028 0.039
Table 4. Basic statistical data for leaves (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.153 0.973 0.015 0 Maximum 0.802 2.309 0.086 0.072 Average 0.372 1.405 0.038 0.030 1.845 RSD 0.175 0.385 0.019 0.022
The concentrations of the individual amino acids alliin, isoalliin and deoxyalliin were also highest in the bulb portion. This was particularly true for alliin. On the other hand, the concentration of methiin was by far the highest in the leaf portion, and lowest in the bulb portion. When these data were subjected to linear discriminant analysis, fourteen of the fifteen varieties tested fell into distinct clusters and could be readily distinguished from each other (Fig. 1).
Fig. 1. Discriminant analysis of garlic parts
Concentrations of sulfur-containing amino acids in different groups of garlic varieties The concentrations of methiin, alliin, isoalliin and deoxyalliin in the flowering plants, semi-bolters and scape absent groups of garlic varieties are presented in Tables 5, 6, & 7.
Table 5. Basic statistical data for flowering plants varieties (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.067 1.023 0.015 0.017 Maximum 0.431 2.587 0.251 0.171 Average 0.241 1.518 0.073 0.082 1.914 RSD 0.103 0.417 0.060 0.048
Table 6. Basic statistical data for scape absent varieties (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.100 0.897 0.022 0 Maximum 0.802 3.666 0.166 0.206 Average 0.356 1.976 0.068 0.086 2.485 RSD 0.210 0.833 0.041 0.062
Table 7. Basic statistical data for semi-bolters varieties (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.084 0.771 0.021 0 Maximum 0.581 2.261 0.164 0.158 Average 0.239 1.480 0.057 0.064 1.840 RSD 0.136 0.475 0.038 0.044
The combined concentration of all four amino acids was far higher in the scape absent group than in either the flowering plants or semi-bolters groups. The concentrations of the individual amino acids alliin and methiin were also far higher in the scape absent group than in the other two groups. The concentration of isoalliin was highest in the flowering plants group and lowest in the semi-bolters group, although the differences were not very large. The concentration of deoxyalliin was highest in the scape absent group and lowest in the semi-bolters group.
Fig. 2. Discriminant analysis of garlic varieties
When these data were subjected to linear discriminant analysis, the varieties fell into clusters which overlapped more than the clusters obtained with the data set indexed by plant portion. Only about half of the forty-five samples could unambiguously be assigned to a particular cluster: seven of the flowering plants samples, ten of the semi-bolters samples, and seven of the scape absent samples (Fig. 2). On the other hand, the clusters were far more distinct if only data on the bulb samples were subjected to linear discriminant analysis, and the data on the stem and leaf samples were disregarded (Tabs. 7, 8 & 9). Fourteen of the fifteen varieties tested fell into distinct clusters and could be readily distinguished from each other (Fig. 3).
Table 8. Basic statistical data for bulbs of flowering plants varieties (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.067 1.561 0.052 0.038 Maximum 0.381 2.587 0.251 0.16 Average 0.210 1.976 0.119 0.083 2.386 RSD 0.115 0.398 0.077 0.047
Table 9. Basic statistical data for bulbs of semi-bolters varieties (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.084 1.473 0.065 0.055 Maximum 0.175 2.261 0.164 0.136 Average 0.130 1.899 0.097 0.081 2.207 RSD 0.033 0.288 0.039 0.033
Table 10. Basic statistical data for bulbs of scape absent varieties (mgg-1) Amino acid methiin alliin isoalliin deoxyalliin total Minimum 0.100 1.502 0.060 0.065 Maximum 0.430 3.666 0.166 0.206 Average 0.272 2.736 0.111 0.144 3.263 RSD 0.147 0.823 0.044 0.054
Fig. 3. Discriminant analysis for bulbs of garlic varieties
CONCLUSIONS The highest concentration of total sulfur-containing amino acids was found in the bulb portion of the garlic plant. Scape absent varieties had significantly higher concentrations of total sulfurcontaining amino acids than either flowering plants or semi-bolters varieties. When the data were indexed by plant portion and subjected to linear discriminant analysis, fourteen of the fifteen varieties tested fell into distinct clusters and could be readily distinguished from each other. When the data were indexed by variety group, the varieties fell into cluster which overlapped more than the clusters obtained with the previous data set. Better separation was obtained when only data from bulb samples was used.
Acknowledgements This study was funded partly by the project COST Action 924, and partly by the Czech Ministry of Agriculture, Project NAZV 1G58084. REFERENCES Hughes J., Tregiva A., Tomsett A.B., Jones M.G., Cosstick R., Collin H.A. 2005. Synthesis of the flavour precursor, alliin, in garlic tissue cultures. Phytochemistry 66: 187-194. Ichikawa M., Ide N., Ono K. 2006. Changes on organosulfur compounds in garlic cloves during storage. J. Agric. Food Chem. 54: 4849-4854.
IPGRI 2001. Descriptors for Allium (Allium spp.).IPGRI Rome, Italy, ECP/GR, AVRDC Taiwan: p. 23, ISBN 92-9043-506-2. Kubec R., Svobodov M., Velek J. 2000. Distribution of S-alk(en)ylcysteine sulfoxide in some Allium species. Identification of a new flavour precursor: S-ethylcysteine sulfoxide (ethiin). J. Agric. Food Chem. 48: 428-433. Velek J., Vos R.D., Schouten A. 1993. HPLC determination of alliin and its transformation products in garlic and garlic-containing phytomedical preparations. Czech J. Food Sci. 11: 445-453. Velek J., Kubec R., Davdek J. 1997. Chemical composition and classification of culinary and pharmaceutical garlic-based products. Z. Lebensm. Unters. Forsch. A 204: 161-164. Velek J., Kubec R., Cejpek K. 2006. Biosynthesis of food constituents: Amino acids: 4. Non-protein amino acids a review. Czech J. Food Sci. 24: 93-109. Yoo K.S., Pike L.M. 1998. Determination of flavor precursor compound S-alk(en)yl-Lcysteine sulfoxides by an HPLC method and their distribution in Allium species. Sci. Hort. 75: 1-10. University of Minnesota, 1999. Available from: http://www.extension.umn.edu/ distribution/cropsystems/components/7317-varieties.html Research Institute of Crop Production, 2005. Available from: http://genbank.vurv.cz/genetic/resources/documents/Allium_2005.pdf ZAWARTO AMINOKWASW SIARKOWYCH W PITNASTU ODMIANACH CZOSNKU Streszczenie Zawarto S-allilo-L-cysteiny (dezoksyalliiny) i sulfotlenkw S-alk(en)ylo-Lcysteiny, w tym alliiny, izoalliiny i sulfotlenku S-metylocysteiny (ang. methiin), wanych nielotnych prekursorw zapachu bya okrelana w cebulach, pdach i liciach 15 rnych odmian czosnku (Allium sativum L.). Analityczna metoda polegaa na wytworzeniu lotnych pochodnych aminokwasw zawierajcych siark z o-ftalodwualdehydo/2-metylopropano-2-tiolu. Pochodne aminokwasw byy okrelane przy uyciu wysokocinieniowej chromatografii cieczowej z odwrcon faz z detektorem UV. Analiza statystyczna uzyskanych wynikw bya wykonana przy uyciu liniowej analizy dyskryminacyjnej, ktra pozwala na scharakteryzowanie i zrnicowanie analizowanych prb.

EFFECT OF PLANT GROWTH REGULATORS ON GROWTH, PHYSIOLOGY AND QUALITY CHARACTERISTICS OF CUCUMIS MELO L.
G. OUZOUNIDOU1, P. PAPADOPOULOU1, A. GIANNAKOULA, I. ILIAS 1 Institute of Food Technology, National Agricultural Research Foundation, Lycovrissi 141 23 Greece, e-mail: geouz@nagref.gr (G.Ouzounidou) Department of Crop Production, Technological Educational Institute of Thessaloniki, PO Box 141, Thessaloniki 541 00 Greece
Summary The experiment was conducted in order to identify the effects of exogenous growth regulators (gibberellic acid (GA3) - 100 M; calcium 3,5-dioxo-4propinoylcyclohexanecarboxylate (prohexadione-calcium) - 100 mgL-1; chlormequat chloride (Cycocel) - 100 mgL-1; 2-chloroethylphosphonic acid (Ethephon) - 100 mgL-1) applied as foliar sprays, on growth, physiology and quality characteristics of Cucumis melo L. cv Galia. GA3 promoted the elongation of the first internode and leaf, while a significant inhibition of the above length under chlormequat chloride and 2-chloroethylphosphonic acid, was observed. The maximum quantum yield of primary photochemistry (Fv/Fm), was significantly decreased under chlormequat chloride, 2-chloroethylphosphonic acid and prohexadione-calcium, showing a possible inhibition of enzymatic process in Calvin cycle. Under the application of gibberellic acid the sugars (fructose, glucose and saccharose) and the soluble solids of melon fruits remained unchanged, as compared to the control, while vitamin C content increased significantly. On the contrary, on exposure to growth retardants, a significant decrease in sugars, soluble solids and vitamin C content and a tendency to increase the titratable acidity of fruits, was induced. The so-called maturity ratio reflecting the ratio of soluble solids to the Acidity, was significantly reduced under prohexadione-calcium, chlormequat chloride and 2-chloroethylphosphonic acid, while under gibberellic acid was increased by 36%, showing a better quality of the fruits due to that hormone. key words: chlorophyll fluorescence, chlormequat chloride, 2-chloroethylphosphonic acid, gibberellic acid, calcium 3,5-dioxo-4-propinoylcyclohexanecarboxylate, vitamin C
INTRODUCTION The manipulation of growth and increasing productivity of plants, is the basis for most plant-related research. Economic studies have demonstrated the marked advances of early fruit production. As a result many techniques are used to accelerate flowering in young plants. Gibberellins (GA) are a family of plant hormones that mediate many responses in plants, from seed germination to senescence (Davies 1995). The most widely available compound is gibberellic acid or gibberellic acid, which induces stem and internode elongation, seed germination, enzyme production during germination and fruit setting and growth (Davies 1995). Plant growth regulators (PGR) are also used in order to control vegetative growth (Latimer 1991). The most commonly used PGR are those which inhibit GA3 biosynthesis. Chlormequat chloride is an onium compound which blocks GA biosynthesis at the step between geranylgeranyl pyrophosphate and copallyl pyrophosphate. Plants treated with this compound, have shortened inernodes and enhanced photosynthesis. Prohexadione-calcium blocks GA biosynthesis between kaurene and kaurenoid acid, whereas 2-chloroethylphosphonic acid is one of the growth retardants which do not inhibit GA biosynthesis. It is reported that 2-chloroethylphosphonic acid decreases photosynthesis like ethylene (Davies 1995). Species vary in responsiveness to PGR and optimum rates may vary with cultivar or growing condition (Latimer 1991). Plant growth retardants generally have the greatest effects on expending or elongating cells, where inhibition of GA synthesis rapidly causes reduction in stem elongation and leaf expansion (Tanimoto 1987). Flowering has been hastened or delayed by PGR depending on species (Latimer 1991). Both the physiological and environmental situations can influence the intensity of chlorophyll fluorescence resulting to changes in some characteristics, such as initial fluorescence (Fo), maximum fluorescence (Fm), and the ratio between them. The chlorophyll fluorescence method, which provides basic information on the functioning of the photosynthetic apparatus, generally compares well with other quality assessment methods (Ouzounidou & Constantinidou 1999). The purpose of this study was to identify the effects of exogenous growth regulators (gibberellic acid, prohexadione-calcium, chlormequat chloride, 2-chloroethylphosphonic acid) applied as foliar sprays, on growth, physiology and quality characteristics of Cucumis melo L. cv. Galia. Galia melon cultivar, is known for its excellent flavour, is widely cultivated in the Northern Greece and has an important market interest. MATERIALS AND METHODS Melon seeds (Cucumis melo L. var. Galia) were sown in 40-cell plug trays filled with a commercial germination mix (Pot ground Klasmann-Dielmann Gmbh.Art-Nr.4390, Germany) and germinated on a greenhouse mist bench set at 22 2oC. At the three to four- leaf stage, uniform seedlings were transplanted
G. OUZOUNIDOU et al. EFFECT OF PLANT GROWTH REGULATORS ... 129 _____________________________________________________________________________________________________
individually and randomly experiment in fifteen experimental plots. Experiment was established on a sandy loam soil whose physicochemical characteristics were silt 18%, clay 5.6%, sand 70.4%, organic matter 0.88%, CaCO3 0.9%, E.C. 1.5 Scm-1, and pH (1:2 H2O) 7.4. Each plant was watered as required and fertilized weekly with 300 ml of nutrient solution containing 60 mg N - 26.2 mg P- 49.8 mg K (water-soluble fertilizer, 20-20-20 F-TOP Ledra Ltd, Thessaloniki) during the experiment. Plants maintained in greenhouse with average day and night temperatures of 302oC and 282oC, respectively, relative humidity 5095%, photosynthetically active radiation of day time maximum intensities at plant height, 500-700 molm-2s-1. The experiments were terminated when plants from each treatment had developed at least one fruit. gibberellic acid at 100 M, chlormequat chloride, 2-chloroethylphosphonic acid and prohexadione-calcium (BAS 125 10W, BASF Corp., Research Triangle Park, N.C) at 100 mgL-1 respectively were evaluated. Each solution contained 0.1% Agral 90 as a surfactant (Syngenta Ltd, UK). A set of 12 melon plants in each plot was sprayed with a low pressure hand-wand sprayer to run off two times at 2-week intervals with each of the above solutions while each plant was treated with 13.9 mL of solutions. First application of the above solution were made 6 weeks after sowing (plants had six to seven leaves). Control plants were treated with water and surfactant. The chlorophyll fluorescence induction curve was monitored by a Plant Efficiency Analyzer (PEA, Hansatech Ltd Kings Lynn, Norfolk, England) (For details see Ouzounidou & Ilias 2005). Chlorophyll a+b was extracted in 100% acetone. Absorbance was measured at 663 and 645 nm using an LKB Ultraspec II spectrophotometer (Ouzounidou & Ilias 2005). The ascorbic acid content of melon was estimated by macerating the fruit sample mechanically with a stabilising agent (5% metaphosphoric acid) and titrating the filtered extract with 2,6 dichlorophenolindophenol. Acidity was measured by titration potentiometrically with alkali 0.1 NaOH to an end-point of 8.2. The acidity was calculated as anhydrous citric acid which is the predominant acid. An automatic digital refractometer of the firm Index Instruments UK, type GPR 12-70, was used to determine the soluble solids of the fruit samples. Results were expressed as Brix at 20oC. Glucose, fructose and saccharose were determined with an HP 1100 Series High Performance Liquid Chromatograph (refractive index detector (RID) using a reverse phase column 250x4 mm of Lichrosphere NH2 bonded to microparticulate silica of 5 m diameter maintained at 37oC. Injection of 20 L of sample solution into a mobile solvent of H2O/AcCN: 25:75 (v/v) with a flow rate of 1.1 mLmin-1 gave the optimum result) (Manolopoulou & Papadopoulou 1998). Fifteen experimental plots (three replications for each treatment) were set up randomly inside the greenhouse using a randomized complete block design. Twelve single plants were used in each treatment combination/replication. Each plot contained three rows with four plants per row spaced 50 cm apart within each row. Distance between rows was 90 cm while distance between plots was 100 cm. Days to anthesis, first internodes length, and length of the first leaf
were recorded. Leaf chlorophyll content, chlorophyll fluorescence, and some quality characteristics were measured of five and three replicate plants from the five treatments. Data were subjected to analysis of variance (ANOVA) and the treatment means were compared using the LSD test at P=0.05, spss procedure. RESULTS Slight but not significant fluctuations on the date of flowering of melon plants, were observed on exposure to gibberellic acid and 2-chloroethylphosphonic acid (Table 1). Gibberellic acid promoted the elongation of the first internode and leaf by 17 and 11% of the control respectively, while a significant inhibition of the above lengths under chlormequat chloride and 2-chloroethylphosphonic acid, was observed (Fig. 1). The maximum quantum yield of primary photochemistry (Fv/Fm) was significantly decreased under chlormequat chloride, 2-chloroethylphosphonic acid and prohexadione calcium showing a possible inhibition of enzymatic process in Calvin cycle. In addition, the significant increase of Tm/Area (which is 100% of the control under prohexadione-calcium) corresponded to disturbances in, or damage to the photosynthetic apparatus (Fig. 2). Under the application of gibberellic acid fructose and glucose and soluble solids content of melon fruits remained unchanged, as compared to the control, while saccharose and vitamin C content decreased and increased significantly, respectively (Table 1, Fig. 3). On the contrary, on exposure to growth retardants, especially to chlormequat chloride and 2-chloroethylphosphonic acid, a significant decrease in sugars, soluble solids and vitamin C content and a tendency to increase the titratable acidity of fruits was induced (Table 1, Fig. 3). The so-called maturity ratio reflecting the ratio of soluble solids to the Acidity, was significantly reduced under prohexadione-calcium, chlormequat chloride and 2-chloroethylphosphonic acid, while under gibberellic acid was increased by 36%, showing a better quality of the fruits due to that hormone (Fig. 3).
Table 1. Effect of gibberellic acid (100), prohexadione-calcium (100 mgL-1), chlormequat chloride (100 mgL-1) and 2-chloroethylphosphonic acid (100 mgL-1) application on time of anthesis of melon plants (n=36) grown in greenhouse conditions and sugars of melon fruits (n=3) Treatment Control Gibberellic acid Prohexadione-calcium Chlormequat chloride 2-chloroethylphosphonic acid Flowering time (days) 62 60 62 62 66 Fructose (% f.w.) 2.3 2.5 2.3 1.6* 1.6* Glucose (% f.w.) 2.4 2.6 2.4 1.7* 1.7* Saccharose (% f.w.) 4.0 3.1* 3.5 0.8* 2.4*
Note: * - differences from control significant at P<0.05
12 First internode First leaf
10
Length (cm)
Plant Growth Regulators
Fig. 1. Changes of first internode and first leaf elongation (n=36) of Cucumis melo L. cv Galia, as a function of PGR application. (For details see Materials and Methods)
3,5 Chla+b Fv/Fm Tm/Area
Chlorophyll concentrationfluorescence parameters
2,5
1,5
0,5
Fig. 2. Changes of chlorophyll a+b concentration (mgg-1 F.W.) and some chlorophyll fluorescence parameters (n=5) of Cucumis melo L. cv Galia fully expanded leaves, as a function of PGR application. (For details see Materials and Methods)
30
Ascorbic acid mg/100g fw Soluble solids % (brix) Soluble solids/ Titratable acidity Index
25
Quality Characteristics
20
15
10
Fig. 3. Changes of some quality characteristics (n=3) of Cucumis melo L. cv Galia fruit, as a function of PGR application. (For details see Materials and Methods)
DISCUSSION It has been claimed that endogenous gibberellic acid play a regulatory role in stem elongation and flowering of some plants (Hisamatsu et al. 2000). First internode and first leaf length of melon plants were significantly enhanced by the application of gibberellic acid; on the contrary, inhibition of growth performance on exposure to the other PGR was observed. Our data are in contrast with the findings of Hisamatsu et al. (2000), where PGR, which inhibit later stages of gibberellic acid biosynthesis, promoted stem elongation and flowering. However, a reduction on plant height under prohexadione-calcium application was found in rice plants with a concomitant reduction of endogenous gibberellin concentration (Nakayama et al. 1992). Prohexadione-calcium has a potential for effective control of vegetative growth in several plant species, however, timing seems to be very important. Multiple low-rate applications are more effective than a single, high-rate treatment (Brown et al. 1997). Inhibition of both cell division and cell elongation has been found under the effect of chlormequat chloride (Rajala & Peltonen-Sainio 2001). 2-chloroethylphosphonic acid, an ethylenereleasing compound, can also be used to retard stem elongation, promote lateral branching, and manipulate flowering date. Since ethylene acts as an antigibberellin (Hayashi et al. 2001), 2-chloroethylphosphonic acid can affect plant height after absorption. Based to our data prohexadione-calcium was found to be milder on growth and maturity ratio as compared to chlormequat chloride and 2-chloroethylphosphonic acid. The relatively low efficacy of prohexadionecalcium may be due to its rapid degradation compared to the others PGR.
Chlorophyll loss is the most obvious feature of the leaf senescence process. Various PGR affect this process. Gibberellic acid has been reported to delay the loss of chlorophyll, but it is unclear whether gibberellic acid acted indirectly on chlorophyll loss by affecting the endogenous cytokinin concentration in the leaves (Jordi et al. 1995). Ethylene is a phytohormone that influences every aspect of plant growth and development. Conflicting results on the effects of ethylene-releasing compounds, like 2-chloroethylphosphonic acid, on net photosynthetic rate (PN) have been reported. It has been shown to increase or decrease PN, but no definite reason has been given for this (Kays & Pallas 1980). In our study not only the chlorophyll a+b concentration but also the chlorophyll fluorescence characteristics were negatively affected by the three PGR application. The efficiency of photochemistry Fv/Fm was declined showing alterations of PSII reactions centers and an inhibition of enzymatic process in the Calvin cycle subjected by prohexadione-calcium, chlormequat chloride and 2-chloroethylphosphonic acid. The quantum yield of electron transport decreased as a result of lowered efficiency of photochemistry and highly reduced state of QA (Ouzounidou et al. 1997). Beside the length reduction, enhanced antioxidant activity following treatment with GA inhibition has been reported in several plants as well as a promotion of leaf pigments has been observed (Rademacher 2000). Our findings are quite different, since under the application of the three growth retardants a significant decrease in the vitamin C content of melon was observed. In parallel, the soluble solids and the maturity ratio was depressed as well. A better quality status of the fruits on exposure to gibberellic acid was detected. Application of PGR on zinnia, impatiens and marigold induced retardation of initial flowering as well as lower plant quality rating compared to the controls (Latimer 1991). These findings are in accordance with ours. On the contrary, malic acid content gradually decreased and soluble solids content slightly increased during maturation under chlormequat chloride, 2-chloroethylphosphonic acid and prohexadione-calcium application in apples (Awad & Jager 2002). However, the differences in plant responses may be due to application methods, plant age, cultivars or growing conditions. Overall, it is obvious that the application of gibberellic acid, prohexadionecalcium, chlormequat chloride and 2-chloroethylphosphonic acid to melon plants leads to different alterations in developmental, physiological and metabolic pathways. Melon fruits seem to improve their quality under gibberellic acid, however plant responses may be due to application methods, cultivars and growing conditions.
REFERENCES Awad M.A., Jager A. 2002. Formation of flavonoids, especially anthocyanin and chlorogenic acid in Jonagold apple skin: influences of growth regulators and fruit maturity. Sci. Hort. 93: 257-266.
Brown R.G.S., Kawaide H., Yang Y., Rademacher W., Kamiya Y. 1997. Daminozide and Prohexadione-Ca have similar modes of action as inhibitors of the late stages of gibberellin metabolism. Physiol. Plant. 101: 309-313. Davies P.J. 1995. Plant hormones, physiology, biochemistry, and molecular biology. Kluwer Academic Publishers, Dordrecht. Hayashi T., Heins R.D., Cameron A.C., Carlson W.H. 2001. Ethephon influences flowering, height, and branching of several herbaceous perennials. Sci. Hort. 91: 305-324. Hisamatsu T., Koshioka M., Kubota S., Fujime Y., King R.W., Mander L.N. 2000. The role of gibberellin biosynthesis in the control of growth and flowering in Matthiola incana. Physiol. Plant. 109: 97-105. Jordi W., Stoopen G.M., Kelepouris K., van der Krieken, W.M. 1995. Gibberellininduced delay of leaf senescence of Alstroemeria cut flowering stems is not caused by an increase in the endogenous cytokinin content. J. Plant Growth Regul. 14: 121-127. Kays S.J., Pallas J.E.Jr. 1980. Inhibition of photosynthesis by ethylene. Nature 385: 51-52. Latimer J.G. 1991. Growth retardants affect landscape performance of zinnia, impatiens, and marigold. HortScience 26: 557-560. Manolopoulou H., Papadopoulou P. 1998. A study of respiratory and physico-chemical changes of four kiwi fruit cultivars during cool storage. Food Chem. 63: 529-534. Nakayama I., Kobayashi M., Kamiya Y., Abe H., Sakurai A. 1992. Effects of a plantgrowth regulator, Prohexadione-Calcium (BX-112), on the endogenous levels of gibberellins in rice. Plant Cell Physiol. 33: 59-62. Ouzounidou G., Moustakas M., Strasser R. 1997. Sites of action of copper in the photosynthetic apparatus of maize leaves: kinetic analysis of chlorophyll fluorescence oxygen evolution, absorption changes and thermal dissipation as monitored by photoacoustic signals. Aust. J. Plant Physiol. 24: 81-90. Ouzounidou G., Constantinidou H.A. 1999. Changes in growth and physiology of tobacco and cotton under Ag exposure and recovery: Are they of direct or indirect nature? Arch. Environ. Contam. Toxicol. 37: 480-487. Ouzounidou G., Ilias I. 2005. Hormone-induced protection of sunflower photosynthetic apparatus against Cu toxicity. Biol. Plant. 49: 223-228. Rademacher W. 2000. Growth retardants: Effects on gibberellin biosynthesis and other metabolic pathways. Ann. Rev. Pl. Physiol. Pl. Mol. Biol. 51: 501-531. Rajala A., Peltonen-Sainio P. 2001. Plant growth regulator effects on spring cereal root and shoot growth. Agron. J. 93: 936-943. Tanimoto E. 1987. Gibberellin-dependent not elongation in Lactuca sativa: recovery from growth retardant-suppressed elongation with thickening by low concentration of GA3. Plant Cell Physiol. 28: 963-973. WPYW ROLINNYCH REGULATORW WZROSTU NA ROZWJ, FIZJOLOGI I CECHY JAKOCIOWE CUCUMIS MELO L. Streszczenie Przeprowadzono dowiadczenie w celu okrelenia wpywu egzogennych regulatorw wzrostu (kwas giberelinowy GA3 (100 M), proheksadion wapnia (100 mg.L-1), chlormekwat chlorek (100 mg.L-1), etefon (100 mg.L-1) stosowanych w formie opryskw dolistnych, na wzrost, fizjologi i cechy jakociowe Cucumis melo L. odm. Galia.
GA3 powodowa wyduanie si pierwszego midzywla i licia na rolinie, podczas gdy chlormekwat chlorek i etefon hamoway wzrost tych czci roliny. Wskanik wzgldnej fluorescencji chlorofilowej (Fv/Fm) by istotnie niszy po zastosowaniu chlormekwat chlorku, etefonu i proheksadionu wapnia, wskazujc na prawdopodobn inhibicj procesw enzymatycznych w cyklu Calvina. Po zastosowaniu GA3, zawarto cukrw (fruktozy, glukozy i sacharozy) i substancji rozpuszczalnych w owocach melona nie zmieniaa si w porwnaniu z kontrol, podczas gdy koncentracja witaminy C istotnie wzrastaa. Z drugiej strony, zastosowanie retardantw wpyno na spadek zawartoci cukrw, substancji rozpuszczalnych i witaminy C oraz wzrost kwasowoci miareczkowej owocw melona. Wspczynnik dojrzaoci owocw, okrelony jako stosunek substancji rozpuszczalnych do kwasowoci, by istotnie niszy po zastosowaniu proheksadionu wapnia, chlormekwat chlorku i etefonu. Natomiast potraktowanie kwasem GA3 spowodowao 35% wzrost tego wspczynnika, wskazujc tym samym na wysz jako owocw melona traktowanych tym hormonem wzrostu.

QUALITY OF PUMPKIN FRUITS IN RELATION TO ELECTROCHEMICAL AND ANTIOXIDATIVE PROPERTIES

Aurelija PAULAUSKIENE1, Honorata DANILCENKO1, Elvyra JARIENE1, Marek GAJEWSKI2, Anna SEROCZYSKA2, Pawe SZYMCZAK2, Aleksandra KORZENIEWSKA2 1 Lithuanian University of Agriculture Studentu g. 11, Kaunas, Lithuania, e-mail: ns@nora.lzua.lt 2 Warsaw Agricultural University Nowoursynowska 166, 02-787 Warszawa
Summary The aim of this work was to investigate electrochemical characteristics and antioxidant activity of pumpkin fruits and to determine the influence of these properties on pumpkin fruits quality. Pumpkins were grown at the experimental field of Lithuanian Agricultural University in Kaunas. Total 11 cultivars of Lithuanian and Polish origin were chosen for the experiment: Stofuntovaja, Bambino, Kroka, emiuina, Miranda, Golosemiannaja, Justynka, Ambar, Amazonka, Karowita, Otylia. Chemical analysis and electrochemical characteristics of pumpkin were performed. Electrochemical parameters were measured in homogenized fresh fruit samples. P value (as combined parameter of pH, redox potential and electrochemical conductivity) was used for evaluation of pumpkins quality. The lowest P value and redox potential in the pumpkins fruit indicate the best quality of Kroka and Miranda cultivars. Pumpkins of this variety are the most suitable for being eaten raw. Fruits of other pumpkin varieties showed significant differentiation of biochemical and electrochemical characteristics. Carotenoids content varied from 0.90 to 11.25 mg100 g-1. Antioxidant activity, expressed as percentage of DPPH inhibition, varied from 12% for emiuina to 89% for Justynka. Relationship between measured quality parameters of fruits was also determined. Strong correlation between total carotenoids content and antioxidative activity was found. key words: antioxidant activity, carotenoids, electrochemical properties, pumpkin, quality INTRODUCTION The quality of vegetables consists of some properties, which can be evaluated using physical and chemical methods (Abbott 1999). One of quality traits is biological activity of a product, and especially its antioxidative activity. Antioxidants are compounds that protect cells against the damaging effects of reac-
tive oxygen species. Some of antioxidant agents are found in vegetables. Products high in vitamin A, vitamin C, vitamin E, and beta-carotene content are believed to be the most beneficial. Carotenoids are red, orange or yellow fatsoluble plant pigments. In human organism, carotenoids play two primary roles: exert antioxidant activity, but some are also converted into the vitamin A. Of the 600 carotenoids that have been identified, about 30 to 50 are believed to have vitamin A activity (Rutkowska 1981). The best known of this group are carotene and -carotene. Experimental studies suggest that a higher dietary intake of carotenoids offers protection against developing certain cancers (e.g., lungs, skin, uterine, cervix, gastrointestinal tract), macular degeneration, cataracts, and other health conditions linked to oxidative or free radical damage (Rock 1997). Special physiological activity of these compounds in human organism as the vitamin A precursors and also as antioxidants, causes increasing interest among researchers in determining their content in different products (Palace et al. 1999). At present, there is no officially recommended dietary intake of carotenoids, but recommended dietary intake of vitamin A is about 1-3 mg per day of retinol equivalent (Murkovic et al. 2002). It is assumed that processing of vegetables improves the bio-availability of carotenoids, since it breaks down the cellulose structure of the plant cell (Van het Hoff et al. 2000). Pumpkins, especially winter squash cultivars (Cucurbita maxima Duch.), are believed to be a good source of carotenoids. The main carotenoids, which are present in winter squash are -carotene and -carotene. Winter squash fruit contains 0.47.5 mg100 g-1 of -carotene and 1.4-8.4 mg100 g-1 of -carotene, depending on cultivar (Bushway 1986, Murkovic et al. 2002, Seroczyska et al. 2006). Other data show that in cultivars of American origin the content of -carotene (in raw fruits) reaches 4.2 mg100 g-1 and of -carotene 0.8 mg 100 g-1 (Holden et al. 1999, Wu et al. 2004). However, USDA reports (Anonymous 2003 a,b) show lower values - 0.8 mg100 g-1 of -carotene and only 0.012 mg100 g-1 of -carotene. For Polish cultivars highest -carotene content (about 12 mg100 g-1) showed cultivar Amazonka, as we described in other report (Sztangret et al. 2004). Investigation of biochemical composition shows the amounts of toxic and harmful materials accumulated in the analyzed products. Electrochemical research methods provide additional information about metabolism and physiological processes. Life processes in plants can be described as chains of electro-chemical or redox reactions. Scientists developed a bio-electrical theory to derive electrical energy value of food from measurements of pH, redox potential and electrical resistance. They suggested that food with low P-value (as combined parameter of three mentioned parameters above), is health promoting (Bloksma et al. 2001). These investigations are carried out in live state of the organism without destruction of cell and reflect the processes taking place in living nature. Application of electrochemical research methods provides the possibilities to evaluate vitality of systems and to enlarge knowledge about plant suitability for food (Danilcenko et al. 2004). The aim of this work was to investigate chemical and electrochemical characteristics of pumpkin fruits and to determine the antioxidant activity of the fruits.
A. PAULAUSKIENE et al. QUALITY OF PUMPKIN FRUITS ... 139 _____________________________________________________________________________________________________
MATERIALS AND METHODS Pumpkins were grown at the experimental field of Lithuanian Agricultural University in Kaunas in 2004-2005. Total 11 cultivars of Lithuanian and Polish origin were chosen for the experiment: Stofuntovaja, Bambino, Kroka, emiuina, Miranda, Golosemiannaja, Justynka, Ambar, Amazonka, Karowita, Otylia. Electrochemical characteristics of pumpkins were determined in Lithuanian University of Agriculture, other characteristics were determined in Warsaw Agricultural University. Dry matter was determined by drying the samples at the temperature of 105C to a constant mass (LST/ISO 751:2000). Ascorbic acid content was determined with titrimetric method (LST/ISO 6557-2:2000). Total carotenoids content was determined with spectrophotometric method (Lichtenthaler & Wellburn 1983). The electrochemical properties of fruit flesh were determined in homogenized fresh fruit samples. The pH value was determined according to LST/ISO 1842:1997, redox potential by ionometer Metrohm AG CH-9101, electrochemical conductivity by conductometer (the conductivity is the inverse of electrical resistance). P-value (as combined parameter of three mentioned parameters) was used for evaluation of pumpkins quality. P value was calculated according to the formula: P = [29.07 mV (rH 2pH)]2 rho1 (W) where: rH redox potential (mV), rho electrical resistance () (Meier-Ploeger & Vogtmann 1988). Antioxidative activity was determined spectrophotometrically, according to Yen & Chen (1995), as the percent of DPPH (2,2-diphenyl-1-picrylhydrazyl) inhibition in fruit flesh methanol extract. Measurements were done after 10 minutes of reaction period, using the wavelength 517 nm. All analyses and measurements were done on representative samples taken in two or three replicates from fresh plant material. Results of the experiment were statistically evaluated with ANOVA program. LSD test was used to show which values differ significantly at P=0.05. Correlation between variables was calculated using Statgraphics Plus v. 4.1 software. Tables show means of the two years of the experiment. RESULTS AND DISCUSSION Pumpkins are sensitive to soil acidity, optimal pH is 6.0-7.5. The soil should be light, quickly warming up and irrigated. Sandy loams and loams with permeable subsoil suit best (Danilcenko et al. 2004). In the field used for the experiment the soil had a neutral pH, was humus-rich, and showed medium content of phosphorus and potassium. Soil analysis data are presented in Table 1.
Table 1. Data of soil analysis for the field used in the experiment pH 6.90 Humus content (%) 3.28 P2O5 content (mgkg-1) 132 K2O content (mgkg-1) 130
Dry matter of fruits was strongly differentiated and varied from 4.74% (for Kroka) to 19.79% (for Ambar) (Table 2). Carotenoids content in fruits was even stronger differentiated and varied from 0.90 mg100 g-1 f.w. (for Miranda) to 11.25 mg100 g-1 f.w. (for Amazonka). High carotenoids content in Amazonka was also reported by Sztangret et al. (2004) and Gajc-Wolska et al. (2005). Antioxidant activity of vegetables is very important from nutritional point of view. The highest activity showed Justynka, but Kroka, Ambar and Amazonka showed also very high activity.
Table 2. Biochemical indices of pumpkin fruits Cultivar Stofuntovaja Bambino Kroka emiuina Miranda Golosemiannaja Justynka F1 Ambar Amazonka Karowita Otylia F1 LSD0.05 Dry matter (%) 9.37 11.37 4.74 6.86 10.93 5.67 13.73 19.79 12.77 9.83 8.26 1.23 Total carotenoids content (mg 100 g-1 f.w.) 2.90 3.45 8.40 1.55 0.90 1.55 10.95 6.31 11.25 6.59 2.32 1.58 Antioxidant activity (% of DPPH inhibition) not determined not determined 73.2 12.1 not determined not determined 88.8 78.5 72.7 49.5 32.2 8.2
For vegetable plant tissue the electrochemical parameter pH is usually used to determine acidity of the flesh. This parameter shows level of activity of protons and expresses energetical aspects. The increase of pH means a loss of vitality in plants (Danilcenko et al. 2005). In the tested samples pH varied from 5.87 to 6.99 and was close to neutral (Table 3). The highest value was found in Kroka pumpkin. It is very important that plant pH is close to human blood pH, which may fluctuate from 7.43 to 7.73. The results show that pH value of Kroka cultivar, equal to 6.99, is most close to pH of human blood. High values of electrical resistance indicate that electrolytes and other cellular ions are more integrated in membranes and cell organelles. Low values indicate free-moving electrolytes, which might be a sign of deterioration of plant cells and tissues (Bloksma et al. 2001). Value of electrical resistance in different samples varied from 77 to 350 ohms (Table 3). The lowest values of resistance were characteristic to Justyna, Amazonka and Ambar pumpkins accordingly 77, 88 and 95 ohms, the highest to Bambino, emiuina
and Miranda accordingly 350, 323 and 314 ohms. Redox potential is other important electrochemical parameter as it reflects the processes of oxidationreduction reactions. In the tested samples the value of redox potential varied from 17.6 mV in Miranda pumpkins to 23.0 mV in emiuina cultivars. The other values varies a little. The obtained data shows that all values of redox potential in the tested pumpkins are lower than 28 mV, which means the domination of reductive environment in the tested samples. When redox potential is low, the plant cells can use freer enthalpy for their activity. Such plants are more suitable for human organism.
Table 3. Electrochemical indices of pumpkin fruits Cultivar Stofuntovaja Bambino Kroka emiuina Miranda Golosemiannaja Justyna Ambar Amazonka Karowita Otylia LSD0.05 pH 6.57 6.97 6.99 6.22 5.87 6.65 6.36 6.44 6.35 6.07 6.15 0.10 Electrical resistance () 245 350 173 323 314 159 77 95 88 118 108 5 Redox potential (mV) 21.8 22.3 18.4 23.0 17.7 20.0 18.0 19.8 18.9 20.3 21.1 0.5 P value (W) 258.9 167.8 94.4 294.3 94.6 237.9 306.5 420.4 373.3 479.4 601.9 0.3
P-value is used to define vitality of the organism and energy distribution tendencies. High and low values represent corresponding entropy of the system. Higher P-value can be interpreted as more openness, while lower values are a sign of ordering or coherence (Bloksma et al. 2001). The lowest P-value was determined in Kroka and Miranda 94.4 and 94.6 W, respectively and the highest in Otylia 601.9 W. Literature indicates that the lower is Pvalue, the more the product is suitable as food and it is healthier from nutritional aspect (Bloksma et al. 2001). In Table 4 correlation coefficients between measured quality parameters of pumpkin fruits is shown. The strongest correlation was found in case of carotenoids content and antioxidative activity of fruit flesh (r = 0.91), and between Pvalue and redox potential (r = 0.96). This is expected result as the redox potential plays a severe role in the P-value formula. However, there was no significant relationship between P value and antioxidative activity of flesh. It is interesting that the correlation between dry matter and carotenoids content in fruits was quite weak (r = 0.45).
Table 4. Correlation coefficients (r) for determined quality parameters of pumpkin cultivars Total Dry Electrical Redox P value Antioxidant carotenoids pH matter resistance potential activity content X 0.45* 0.51* 0.03 0.47 0.08 0.70** X 0.30 0.44 0.11 0.25 0.91** 0.30 X 0.48* 0.96** 0.42 0.61* 0.44 0.48* X 0.57 0.24 0.23 0.11 0.96** 0.57 X 0.54* 0.37 0.25 0.42 0.24 0.54* X 0.17 0.91** 0.61* 0.23 0.37 0.17 X
Dry matter Carotenoids 0.45* content pH 0.51* Electrical 0.03 resistance Redox potential 0.47 P value 0.08 Antioxidant 0.70** activity
Note: **means significant correlation at P=0.01, *means significant correlation at P=0.05
CONCLUSIONS 1. Investigated pumpkin cultivars differ in chemical and electrochemical characteristics, concerning dry matter and carotenoids content, pH of fruit flesh, electrical resistance, redox potential and P-value. Therefore, nutritional and biological value of pumpkin differs greatly. 2. Kroka and Miranda cultivars show the lowest P-value, which is a combined parameter taking into account different electrochemical properties of fruit flesh. 3. There is strong and significant relationship between carotenoids content in pumpkin fruits and their antioxidative activity, and between pH of fruit flesh and redox potential.
Note: Research was supported by Lithuanian State Science and Studies Foundation. REFERENCES Abbott J. 1999. Quality measurements of fruits and vegetables. Postharvest Biol. Technol. 15: 207-225. Anonymous 2003a. Winter squash. USDA Database for Standard Reference, Release 16. http//www.nal.usda.gov. Anonymous 2003b. USDANCC Carotenoid Database for U.S. Foods, 20. http//www.nal.usda.gov. Bloksma J., Northolt M., Sluber M. 2001. Parameters for apple quality. Louis Bolk Institute: 78-81. Bushway R.J. 1986. Determination of - and -carotene in some raw fruits and vegetables by high-performance liquid chromatography. J. Agric. Food Chem. 34: 409-412.
Danilcenko H., Jarien E., Paulauskiene A., Kulajtiene J., Viskelis P. 2004. The influence of fertilization on quality and chemical composition of pumpkin. Annales UMCS. Lublin. Sec. E. 59 (4): 1949-1956. [in Polish]. Danilcenko H., Paulauskien A., Rutkovien V., Kulaitien J. 2005. The influence of various fertilizers on electrochemical properties of pumpkins fruits. Sodininkyst ir darininkyst. Mokslo darbai 24 (3): 78-86. Holden J.M., Eldridge A.L., Beecher G.R., Buzzard M., Bhagwat S., Davis C.S., Douglass L.W., Gebhardt S., Haytowitz D., Schakel S. 1999. Carotenoid content of U.S. foods: an update of the base. J. Food Comp. Anal. 12, 169-196. Gajc-Wolska J., Gajewski M., Radzanowska J., Niemirowicz-Szczytt K., Korzeniewska A. 2005. The fruit quality of chosen hybrids and cultivars of pumpkin (Cucurbita maxima Duch.). Veg. Crops Res. Bull. 63: 34-43. Lichtenthaler H.K., Wellburn A.R., 1983. Determination of total carotenoids and chlorophyls a and b of leaf extracts in different solvents. Biochem. Soc. Transact. 603: 591-592. LST ISO 1842. 1997. Fruit and vegetable products Determination of pH. LST ISO 751. 2000. Fruit and vegetable products Determination of water-insoluble solids. LST ISO 6557-2:2000. Fruit, vegetable and their products Determination of ascorbic acid content. Part 2. Standard Methods. Meier-Ploeger A., Vogtmann (Hrsg.) H. 1988. Lebensmittelqualitat ganzheitliche Methoden und Konzepte. Karlsruhe, Mueller: 67-86. Murkovic M., Muelleder U., Neunteufl H. 2002. Carotenoid content in different varieties of pumpkins. J. Food Comp. Anal. 15: 633-638. Palace V.P., Khaper N., Qin Q., Singal P.K. 1999. Antioxidant potentials of vitamin A and carotenoids and their relevance to heart disease. Free Rad. Biol. Med. 25: 746761. Rock C.L. 1997. Carotenoids: biology and treatment. Pharmacol. Ther. 75: 185-97. Rutkowska U. 1981. Witamina i prowitamina A. In: Wybrane metody analizy skadnikw ywnoci i wartoci odywczej. PZWL Warszawa: 302-331. [in Polish] Seroczyska A., Korzeniewska A., Sztangret-Winiewska J., Niemirowicz-Szczytt K., Gajewski M. 2006. Relationship between carotenoids content and flower or fruit flesh colour of winter squash (Cucurbita maxima Duch.). Folia Hortic. 18/2: 51-61. Sztangret J., Korzeniewska A., Horbowicz M., Niemirowicz-Szczytt K. 2004. Comparison of fruit yields and carotenoids content in new winter squash hybrids (Cucurbita maxima Duch.). Veget. Crops Res. Bull. 61: 51-60. Van het Hoff K.H., de Boer B.C., Tijburg J.B.M., Lucius B.R.H.M., Zijp I., West C.E., Hautvast J.G.A.J., Westrate J.A. 2000. Carotenoid bioavailability in humans from tomatoes processed in different ways determined from the carotenoid response in the triglyceride-rich lipoprotein fraction of plasma after a single consumption and in plasma after four days of consumption. J. Nutr. 130: 1189-1196. Wu X., Gu L., Holden J., Haytowitz, Gebhardt S.E., Beecher G., Prior R.L. 2004. Development of a database for total antioxidant capacity in foods: a preliminary study. J. Food Comp. Anal. 17: 407-422. Yen G.C., Chen H.Y. 1995. Antioxidant activity of various tea extracts in relation to their antimutagenicity. J. Agric. Food Chem. 43: 27-32.
JAKO OWOCW DYNI W ZALENOCI OD WACIWOCI ELEKTROCHEMICZNYCH I ANTYOKSYDACYJNYCH Streszczenie Celem pracy byo zbadanie waciwoci elektrochemicznych i aktywnoci antyoksydacyjnej owocw dyni oraz okrelenie wpywu tych waciwoci na jako owocw. Dynie uprawiano na polu dowiadczalnym Uniwersytetu Rolniczego w Kownie. Do dowiadczenia wzito 11 odmian pochodzcych z Litwy i z Polski: Stofuntovaja, Bambino, Kroka, emiuina, Miranda, Golosemiannaja, Justynka, Ambar, Amazonka, Karowita, Otylia. Analizy chemiczne i pomiary elektrochemiczne przeprowadzono wedug standardowych metod. Parametry elektrochemiczne okrelano w homogenizowanych prbkach miszu owocw. Jako wskanik jakoci owocw by uyty obliczony parametr P (kombinacja wartoci pH, potencjau redoks i przewodnictwa elektrycznego). Najnisz wartoci P i potencjau redoks charakteryzoway si odmiany Kroka i Miranda. Wskazuje to na najwysz jako owocw tych odmian i ich dobr przydatno do spoycia w stanie surowym. Owoce innych odmian wykazay due zrnicowanie badanych cech. Zawarto karotenoidw ogem wahaa si od 0,90 to 11,25 mg100 g-1. Aktywno antyoksydacyjna, wyraona jako procent inhibicji DPPH, wahaa si od 12% dla odmiany emiuina do 89% dla odmiany Justynka. Okrelono rwnie zalenoci midzy badanymi cechami owocw dyni. Midzy innymi wykazano istotn korelacj midzy ogln zawartoci karotenoidw a aktywnoci antyoksydacyjn owocw.

CHEMICAL COMPOSITION AND SENSORY QUALITY OF DRY AND FROZEN HERB OF DILL (ANETHUM GRAVEOLENS L.)
Janina GAJC-WOLSKA, Wiesawa ROSON, Ewa OSISKA Department of Vegetable and Medicinal Plants Warsaw Agricultural University Nowoursynowska 159, 02-776 Warsaw, Poland
Summary Two cultivars of dill (Szmaragd and Moravan) were used for this experiment. The seeds were sown two times: in the second ten days of May and in the second ten days of June. The raw material (shoots with leaves) was harvested in first ten days of July and in first ten days of August. After harvest the raw material was dried in the laboratory drying chamber at 35C, and stabilized by deep freezing at minus 85C for 48 hours. Then it was kept at -20C for two months. Two-years results of the study indicate that the content of volatile oil as well as overall odour intensity and intensity of the dill odour were affected by the method of stabilization, time of harvest and cultivar. The main component of the volatile oil from dry raw material was -phellandrene, whereas from deep frozen herb dill ether and p-cymene. The high overall odour intensity and the positive note of the dill odour were characteristic for the frozen raw material of the cultivar Szmaragd from the first harvest and from the cultivar Moravan from the second harvest as well as the dried raw material of the cultivar Szmaragd from both harvests. key words: cultivars, term of harvest, volatile oil, sensory analysis INTRODUCTION Dill (Anethum graveolens L.) has been long known in the Polish cuisine as a common and valuable spice vegetable. Usually, for culinary purposes, fresh young leaves are used or entire plants, collected at the beginning of fruit ripening. Each part of this plant contains volatile oil, whose amount and chemical composition determine the quality of the material (Gra & Wooszyn 2001). Under climatic conditions of Poland, fresh dill herb is available from May to October. In other months the seasoning appears in the market in a processed form - dried, salted or frozen. During preservation of the material special attention is taken for the quality of the product, particularly to its appearance, smell and taste (Lisiewska & Kmiecik 1997). Chemical composition is also an im-
portant quality indicator of an herba, as it affects its nutritive and dietetic values (Abbott 1999, Justesen & Knuthsen 2001). The application of a proper preservation method enables the maintainenance of a high quality product and minimize physical, chemical and sensory changes. The aim of the study was to analyse chemical composition and sensory evaluation of dill herb (leaves and shoots) (Anethum graveolens L.) preserved through drying and freezing. MATERIAL AND METHODS The experiment was conducted in 2002 and 2003 on the Experimental Field, in the Laboratory of Chemical Analysis, and in the Sensory Laboratory of Vegetable and Medicinal Plants Department Warsaw Agricultural University. The material studied was comprised of two dill varieties: Szmaragd and Moravan. Szmaragd is a medium late, high-yielding variety. The plants are erect, with very leafy stems, slightly covered with waxy cutin. Owing to high content of volatile oils the herb is very aromatic. Moravan is a variety of dark green foliage and strong aroma. The plants have good growing capacity, bloom late. The variety well suited for drying and freezing. Dill was sown on two dates: in the second half of May and in the second decade of June, on plots of a 5 m2 area, in four replicates. On each plot 12.5 g of seeds were sown. The material (shoots and leaves) was collected in the first decade of July (I period of harvest) and in the first decade of August (II period of harvest). Average temperatures and sums of rainfall are given in Table 1. After harvesting, the material was stabilized either by drying at 35C or by deep-freezing at -85C for 48 hours, and stored at the temperature maintained at -20C. After three moths the material was subjected to chemical and sensory analyses. The content of volatile oil was extracted with vapour distillation, according to F.P. VI. The chemical compound of the oil was analysed with a gas chromatography method using an Anglia Instrument gas chromatograph at the following separation conditions: detector temperature 250C; inlet chamber temperature 220C; program oven temperature: initial temperature 60C, a 4C temperature gain per minute, 220C maintained for five minutes; a capillary column - Carbowax 20M (length 25 mm, 0.32 mm); sample size 0.32 mm; carrier gas helium. Sensory analysis concerned the evaluation of dill herb profile odour, its intensity and overall intensity. Sensory profile evaluation followed the methodology given in Polish Standard PN ISO 11035. The samples of 0.5 g of dried dill and 5 g of frozen dill were taken to the estimation. The selection of attributes applied, definitions and sequencing of particular odour notes appearing was estimated by nine panellists. Overall odour of dill herb and the intensity of particular attributes were estimated with the use of a ten centimetre, non-structural linear scale. Evaluations plotted on the scale had been then converted into numerical values and expressed in conventional units of 1-10 scale. The obtained results had been statistically analyzed using Three-Way Analysis of Variance.
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Differences between sources of variation were defined with Fisher-Snedecor test at P=0.01 and P=0.05. Tuckey test was used for detailed comparison between means. RESULTS AND DISCUSSION Among many factors affecting the content of volatile oil in dill herb, the method of its stabilization is of great importance (Venskutonis 1997, Barbieri et al. 2004). Results of the present study indicate that the amount of volatile oil was significantly higher in dried material as compared to the frozen one. The content of this compound appeared to depend also on harvest time and on the particular variety. The material collected in the first decade of August showed higher content of the oil as compared to the material collected at the beginning of July. Moreover, significantly higher amount of the oil was found in Szmaragd plants compared to the variety Moravan(Table 1). The quality of the investigated material was strictly related to fractions of different chemical compounds in volatile oil. According to Gra & Wooszyn (2001) such compounds as -phellandrene, p-cymene, limonene and dill ether (3,6-dimethyl-2,3,3a,4,5,7a-hexahydro-1-benzofuran) are specific for dill herb. The results of the study allow a comparison of stabilization methods responsible for chemical compounds composition in the oil. Both in the first and in the second harvests, dried material showed higher content of -phellandrene as compared to frozen material. The content of dill ether and p-cymene was in both dates higher in frozen material comparing to the dried one. The amount of limonene was similar in dried and frozen material, irrespective of the date of harvest (Table 2).
Table 1. Content of volatile oil (mL.100 g-1) (mean for 2002-2003) I period of harvest II period of harvest Form of dill herb Szmaragd Moravan Szmaragd Moravan Dried 1.03 b 0.93 c 1.12 a 1.13 a Frozen 0.10 f 0.10 f 0.23 d 0.15 e Mean for period 0.54 b 0.66 a of harvest Mean for Szmaragd 0.62 a varieties Moravan 0.58 b Mean for processing method 1.05 a 0.14 b
Note: means marked with the same letter do not differ significantly (P=0.05)
Table 2. Content of identified constituents of essential oil from dill herb (%) (mean for 2002-2003)
Constituent (% in essential oil) Form p - phel - phelVariety of dill liDill cyme landrene landrene herb pinene pinene monene ether ne I period Szmaragd dried 3.01 58.78 8.24 17.39 5.68 of frozen 1.73 0.03 23.99 9.96 7.97 33.62 15.26 harvest Moravan dried 2.79 55.14 18.22 10.93 7.48 frozen 1.92 37.45 13.86 24.23 12.44 II peSzmaragd dried 3.10 0.04 66.60 17.17 7.57 4.35 riod of frozen 2.28 36.65 13.78 12.97 27.13 16.00 harvest Moravan dried 3.55 0.07 53.14 15.69 9.49 9.07 frozen 2.36 46.36 12.12 26.27 13.58 Period of harvest
mirist caricin vone 0.56 1.75 0.79 1.75 0.11 1.57 0.39 -
Overall odour intensity and dill odour intensity are primary sensory attributes for evaluating dill herb quality. The characteristic odour note of dill is affected by the content of volatile oil as well as its chemical composition (Blanc & Grosh 1991, Pino et al. 1995). The present study proved that such factors as the method of material stabilization, date of harvest as well as variety significantly influence both the overall odour intensity and dill odour intensity. Dried material showed both higher overall odour intensity and dill odour intensity as compared to frozen material. Slight difference in the overall and dill odour intensity between dried and frozen material results from the fact that the amount of the material used for the analysis was in both cases equal when expressed as dry matter. Dill material collected in the second date obtained slightly higher marks of odour intensity for both odour attributes comparing to the material collected in the first date. Szmaragd showed a significantly higher intensity of dill odour than Moravan (Table 3 & 4).
Table 3. Overall intensity of dill herb odour (scale 0-10) (mean for 2002-2003) I period of harvest II period of harvest Form of dill herb Szmaragd Moravan Szmaragd Moravan Dried 6.30 b 4.10 d 7.90 a 6.70 b Frozen 5.60 c 5.40 c 5.00 cd 3.20 e Mean for period 5.35 b 5.70 a of harvest Mean for Szmaragd 6.20 a varieties Moravan 4.85 b Note: see Table 1 Mean for processing method 6.25 a 4.80 b
Table 4. Intensity of dill odour (scale 0-10) (mean for 2002-2003) Form of dill I period of harvest II period of harvest herb Szmaragd Moravan Szmaragd Moravan Dried 5.10 b 3.60 d 6.90 a 4.70 c Frozen 4.20 c 3.00 e 3.40 d 1.20 f Mean for period 3.98 b 4.05 a of harvest Mean for Szmaragd 4.90 a varieties Moravan 3.13 b Note: see Table 1 Mean for processing method 5.07 a 2.95 b
For profile odour evaluation of the material studied, ten attributes, potentially contributing to the overall odour note of dill herb had been chosen. Such attributes as dill odour, herbal odour and chamomile odour had been classified as positive, whereas sharp odour, acidic odour, chemical odour, hay odour, fungous odour and earthy odour had been regarded negative. The PCA graph, presenting the results of a profile evaluation, points to the differentiation of the dry material sensory quality in relation to the variety and the date of harvest. Szmaragd II (II period of harvest) showed high values of overall odour intensity and dill odour intensity, however, the presence of some negative traits as chemical odour, sharp odour and earthy odour slightly reduced the quality of the material. High note of dill odour was also obtained in Szmaragd I sample (I period of harvest). However, in his case a considerable intensity of acidic odour significantly deteriorated the quality of this material. The fact that samples of Moravan I (I period of harvest) and Moravan II (II period of harvest) were localized close to such attributes as fungous odour, hay odour and strange odour indicates a low sensory value of this material (Fig. 1). Processing the data of profile evaluation of frozen dill with the PCA method allowed us to find out that Szmaragd I sample (I period of harvest) shows high overall odour intensity and high note of dill odour. However, close localization of the latter to such odour attributes as hay odour and off odour may indicate a deteriorated sensory value of the material. Moravan II (II period of harvest) showed high intensity of dill odour but the quality of the material had been affected by a negative note of acidic odour. The localization of Szmaragd II (II period of harvest) and Moravan I (I period of harvest) samples close to the attributes of negative traits (chemical odour, grasses odour, acidic odour and earthy odour) points at a low sensory value of this material (Fig. 2).
2,7 4 1,7 PCA (61.51%) Moravan I 1 0,7 9 8 11 3 Moravan II -1,3 5 -2,7 -1,7 6 Szmaragd I 1,3 2,3 3,3 2 7 10 Szmaragd II
-0,3
-2,3
-0,7 0,3 PCA (30.15%)
Fig. 1. PCA projection of differences and similarities of sensory quality of dried dill herb varietys Szmaragd I, Moravan I (I term of harvest) and Szmaragd II, Moravan II (II term of harvest). Attributes evaluated: 1-overall intensity of odour, 2-sharp odour, 3-dill dried odour, 4-herbal odour, 5-chamomile odour, 6-acidic odour, 7-chemical odour, 8-hay odour, 9-fungous odour, 10-earthy odour, 11-off-odour
2,8 3 1,8 PCA (57.53%) 5 Moravan II 7 4 8 Moravan I 2 Szmaragd II -2,2 -2,8 -1,8 -0,8 PCA (27.77%) 0,2 1,2 2,2 3,2 6 9 1 Szmaragd I
0,8
-0,2
-1,2
Fig. 2. PCA projection of differences and similarities of sensory quality of frozen dill herb varietys Szmaragd I, Moravan I (I time of harvest) and Szmaragd II, Moravan II (II time of harvest). Attributes evaluated: 1-overall intensity of odour, 2-sharp odour, 3-dill odour, 4-grasses odour, 5-acidic odour, 6-chemical odour, 7-hay odour, 8-earthy odour, 9-off-odour
CONCLUSIONS High concentration of volatile oil as well as the overall odour and dill odour intensity in dill material depend on the method of material stabilization applied, the date of harvesting and the variety. Stabilization methods significantly affected the share of particular volatile oil compounds. -phellandrene predominated in dry material while dill ether and p-cymene in frozen material. The content of limonene was similar irrespectively the stabilization method. The intensity of both the positive odour notes (attributes) like the overall odour, dill odour and the negative odour notes like sharp odour, hay odour, chemical odour and fungous odour are of key importance in determining the sensory value of the frozen and dried material of dill. Among assessed samples of dried material, high sensory value was represented by Szmaragd samples collected from both harvest dates. In the case of frozen material the value was the highest for Szmaragd samples from the first harvest date and Moravan samples from the second date. The best method of material stabilization for ,Szmaragd variety was drying.
REFERENCES Abbott J.A. 1999. Quality measurement of fruit and vegetables. Postharvest Biol. Technol. 15: 207. Barbieri S., Elustondo M., Urbicain 2004. Retention of aroma compounds in basil dried with low pressure superheated steam. J. Food Engineering 65: 109-115. Blanc W., Grosch W. 1991. Evaluation of Potent Odorants in Dill Herb (Anethum Graveolens L) by Aroma Extract Dilution J. Food Sci. 56 (1): 63-67. Gra J., Wooszyn A. 2001. Olejek kopru ogrodowego. Aromaterapia 7 (3): 5-12. [in Polish] Justesen U., Knuthsen P. 2001. Composition of flavonoids in fresh herbs and calculation of flavonoid intake by use of herbs in traditional Danish dishes. Food Chem. 73: 245-250. Lisiewska Z., Kmiecik W. 1997. Effect of freezing and storage on quality factors in Hamburg and leafy parsley. Food Chem. 60(4): 633-637. Pino J.A., Rosado A., Goire L., Roncal E. 1995. Evaluation of flavour characteristic compounds in dill herb essential oil by sensory analysis and gas chromatography. J. Agric. Food Chem. 43(5): 1307-1309. Venskutonis P.R. 1997. Effect of drying on the volatile constituents of thyme (Thymus vulgaris L.) and sage (Salvia officinalis L.). Food Chem. 59(2): 219-227.
SKAD CHEMICZNY I JAKO SENSORYCZNA SUCHEGO I WIEEGO ZIELA KOPRU OGRODOWEGO (ANETHUM GRAVEOLENS L.) Streszczenie Do bada wzito dwie odmiany kopru ogrodowego Szmaragd i Moravan. Nasiona wysiano w dwch terminach: w drugiej dekadzie maja i czerwca. Ulistnione pdy zbierano w dwch terminach: w pierwszej dekadzie lipca i sierpnia. Po zbiorze suro-
wiec utrwalono stosujc metod suszenia w temperaturze 35C oraz gbokie mroenie w temperaturze -85C przez 48 godzin, a nastpnie utrzymujc temperatur na poziomie -20C. Dwuletnie wyniki bada wskazuj, e zawarto olejku eterycznego jak rwnie oglna intensywno zapachu i intensywno zapachu kopru zaleay od terminu zbioru metod stabilizacji oraz odmiany. Metody utrwalania istotnie wpyny na udzia pojedynczych skadnikw olejku eterycznego. W surowcu suchym dominowa felandren, a w surowcu mroonym dill eter i p-cymen. Wysokie noty oglnej intensywnoci zapachu i zapachu kopru uzyska surowiec mroony odmiany Szmaragd zbierany w pierwszym terminie i odmiany Moravan zbierany w terminie drugim, jak rwnie surowiec suchy odmiany Szmaragd zbierany w obu terminach.

EFFECT OF POSTHARVEST IRRADIATION ON HEALTH AND SENSORY RELATED PROPERTIES OF APPLES AND BROCCOLI
Gunnar B. BENGTSSON1, Sidsel Fiskaa HAGEN1,2, Grethe Iren A. BORGE1, Roman SCHNER1, Emanuele LOMBARDO1, Jennifer SCHNER1, Wolfgang BILGER3, Arvid BERGE2, Karin HAFFNER2, Knut Asbjrn SOLHAUG2 1 Matforsk AS Norwegian Food Research Institute, Osloveien 1, NO-1430 Aas, Norway 2 Norwegian University of Life Sciences, Department of Ecology and Natural Resource Management, and Department of Plant and Environmental Sciences, NO-1432 Aas, Norway 3 University of Kiel, Botanical Institute, Am Botanischen Garten 1-9, DE-24118 Kiel, Germany
Summary Flavonoids and chlorogenic acid can be induced postharvest in apple skin. However, the effect of postharvest irradiation on product quality in general has been investigated in fresh fruit and vegetables to a very limited extent. As examples we have studied the effects of post-harvest radiation treatments on several parameters of apples and broccoli related to taste and human health. Norwegian grown broccoli and apples were treated 10-12 days with various combinations of visible light and UV radiation in controlled climate at 5C and 10C, respectively. L-ascorbic acid, chlorogenic acid and flavonoids were quantitated by HPLC, and total phenols, soluble solids and titratable acidity by other methods. Antioxidant capacity was measured in methanol extracts by the ORAC (Oxygen Radical Absorbance Capacity) method. In broccoli, the levels of flavonols and the antioxidant capacity were much higher in flower buds than in stalks. The natural variation between broccoli plants was very large in flower buds: tenfold for flavonol content and twofold for antioxidant capacity. This variation was not reduced by the radiation treatment. Flavonol levels in flower buds tended to increase after treatment with visible light + UV-A + UV-B, but the increase was just outside the border of statistical significance. Radiation treatment did not change the antioxidant capacity in broccoli. Treatment of apples with UV-B and visual light increased red colour, antioxidant capacity and the contents of flavonols, anthocyanins, chlorogenic acid and ascorbic acid in the peel, but not in the flesh. The effect was strongest in suboptimal (green) apples harvested from the inner tree canopy. The only changes after treatment with visible light were increases in quercetin glycosides and ascorbic acid in the skin of green apples. There was no influence on the contents of catechins, pro-
cyanidins, phloridzin, total phenols, soluble solids or titratable acidity in apples after any of the treatments. The study shows that it is possible to increase not only the contents of specific phenolic constituents but also other health-related properties in apples by postharvest irradiation, while at the same time sensory related properties did not change. The broccoli experiment was probably carried out at sub-optimal conditions and the number of properties studied was small. The fact that controlled radiation treatment postharvest can change the health-related quality is an indication that also the incident light during distribution of fruit and vegetables could have an effect, which might be positive or negative depending upon the conditions. key words: Brassica oleracea L. var. italica, Malus domestica Borkh, UV irradiation, postharvest, flavonoids, ascorbic acid, ORAC, flavour INTRODUCTION Ultraviolet (UV) radiation from the sun can damage living cells, and therefore there exists a defence mechanism in plants where radiation at certain wavelengths induces the formation of UV-absorbing flavonoids in the epidermis. In addition to colourless flavonoids such as flavones and flavonols, anthocyanins with a red colour are synthesised in response to the sun. There is extensive epidemiological evidence that a high consumption of fruit and vegetables is associated with a reduced risk of various illnesses (World Cancer Research Found 1997). Probably this is connected with the presence of health beneficial constituents in addition to nutrients, for instance flavonoids, carotenoids and glucosinolates. The levels of health-related constituents in plant products are determined by the genome, as well as by the conditions during growth, harvest and postharvest storage and treatment. It has been earlier shown that some flavonoids and chlorogenic acid can be induced postharvest in apple skin by irradiation (e.g. Arakawa et al. 1985, Lancaster 2000, Ubi et al. 2006). However, the effect of postharvest irradiation on product quality in general has been investigated in fresh fruit and vegetables to a very limited extent. As examples we have studied the effects of irradiation with UV and visible light on apples and broccoli on several parameters related to sensory quality and human health. MATERIALS AND METHODS Norwegian grown broccoli (Brassica oleracea L. var. italica, cv. Marathon) or apples (Malus domestica Borkh. cv. Aroma) were treated with various combinations of visible light (Vis) and UV radiation in controlled climate rooms. Broccoli heads were stored in vases for 12 days at 5C with the following radiation treatments 6 h per day: Vis, Vis + UV-A and Vis + UV-A + UVB. This was achieved by using appropriate lamps and filters with selective
G.B. BENGTSSON et al. EFFECT OF POSTHARVEST IRRADIATION ... 155 ____________________________________________________________________________________________________
transparency giving an approximate irradiance of 70 molm-2s-1 for PAR and 0.5 Wm-2 for UV radiation (Bengtsson et al. 2006). Similarly, apples were stored for 10 days at 10C and 95% relative air humidity with radiation treatment 12 h per day: Vis and Vis + UV-B, with an irradiance of 30 molm-2s-1 for PAR and 0.20 Wm-2 for UV radiation. Respective control samples were stored in the dark in the same rooms. Flavonoid aglycons were measured by high-performance liquid chromatography (HPLC) in flower buds, floret stalks and the main stem of the broccoli heads after acidic hydrolysis of methanol extracts (Bengtsson et al. 2006). Skin and flesh samples of apples were separated after freezing tissue plugs in liquid nitrogen, and L-ascorbic acid, chlorogenic acid and flavonoids were quantitated by HPLC (Hagen et al. 2006). Total phenols were measured by the FolinCiocalteu method. In apple juice from edible tissue (skin and flesh), soluble solids were measured by refractometry and titratable acidity by titration with 0.1 M NaOH to pH 8.1. Total antioxidant capacity was measured in methanol extracts by the Oxygen Radical Absorbance Capacity (ORAC) method (Cao et al. 1999). The storage kinetics of epidermal absorbance in apple skin at 375 and 470 nm, correlating to quercetin glycosides and anthocyanins, respectively, was measured by a non-destructive method (Bilger et al. 2001), using UV-A-PAM Chlorophyll Fluorometer (Gademann Instruments, Wrzburg, Germany) and PAM-2000 Fluorometer (Walz, Effeltrich, Germany) as described elsewhere (Hagen et al. 2006). RESULTS The postharvest treatment of broccoli heads and apples did not change the apparent quality; the products were considered to be of good market value after treatment. The natural variation between broccoli plants was very large: for flavonol content tenfold and for the ORAC value more than twofold in dark green flower buds. This variation was not reduced by the post-harvest radiation treatment. The levels of flavonols and total antioxidant capacity were much higher in flower buds than in stalks of the inflorescence or in the stem (Table 1). Flavonol levels in flower buds tended to increase after treatment with visible light + UVA + UV-B, but the change was just outside the border of being statistically significant (Table 2). Radiation treatment did not change the total antioxidant capacity (Table 2). The correlation between flavonoid contents and total antioxidant capacity was weak: r=0.37, p<0.05 (Fig. 1).
Table 1. Distribution of flavonoids in broccoli heads (mg100 g-1 fresh matter, mean SD) Flavonoid Quercetin (Q) Kaempferol (K) Q+K Flower buds (n=8) 3.32 1.45 13.0 7.5 16.3 8.7 Floret stalks (n=8) 0.24 0.17 0.83 0.84 1.07 0.99 Stem (n=2) 0.10 0.11 0.21
Table 2. Effect of postharvest irradiation on flavonoid contents (mg100 g-1 fresh matter) and antioxidant capacity, ORAC (mol Trolox eq. 100 g-1 fresh matter) in broccoli flower buds (mean SD, n = 7-8) Parameter Quercetin (Q) Kaempferol (K) Q+K ORAC Control (dark) 4.67 1.97 17.0 9.9 21.6 11.7 20.1 3.7 Vis 5.73 2.85 24.8 13.0 30.5 15.8 21.5 6.5 Vis + UV-A 5.38 1.56 18.2 7.0 23.6 8.0 19.3 3.8 Vis + UV-B 6.16 1.77 26.0 12.9 32.1 14.6 20.6 5.4
35
r = 0,37 (p<0,05)
ORAC
25
15
10
20
30
40
50
60
K+Q
Fig. 1. Relation between antioxidant capacity (ORAC) and content of flavonols (K+Q = kaempferol + quercetin) in broccoli flower buds. Broccoli heads from all postharvest treatments are included. The units are: ORAC value (mol Trolox eq./g) and flavonols (mg aglycons100-1 g), fresh matter basis
Postharvest irradiation of apples changed the sum of flavonoids significantly with Vis + UV-B and only in the peel of green, suboptimal apples according to the wet chemistry data (Fig. 2). There were no changes in the flesh of any parameter. Treatment with Vis + UV-B increased red colour (data not shown), antioxidant capacity and the content of flavonols (quercetin-3glycosides), anthocyanins (represented by the predominant cyanidin-3galactoside), chlorogenic acid and L-ascorbic acid in the peel of the green apples (Table 3). Radiation treatment, or storage in the dark, had no influence on the content of (-)epicatechin (the predominant catechin), procyanidins, phloridzin, total phenols, soluble solids or titratable acidity in the peel of any of the apples. Of the parameters measured by wet chemistry in red apples, only chlorogenic acid and antioxidant capacity showed significant increases in the peel, from 6.5 to 17.9 mg100 g-1 and from 9.1 to 13.7 mol Trolox eq. per 100 g fresh matter.
Red apples
250
. Flavonoids (mg 100 -1 g fresh matter)
Green apples
Before light treatment After light treatment
200
150
***
100
50
0
PAR + UV, PAR apples +red UV-B PAR, red PAR apples Control, C red apples PAR + UV, PAR green + UV-B apples PAR, green PAR apples Control, C green apples
***) significant at P0.01

Fig. 2. Effect of postharvest treatment of whole Aroma apples with visible or visible + UV-B radiation on flavonoid content in peel
With the non-destructive method using chlorophyll fluorescence small changes in epidermal absorbance in the red apples were statistically significant: 12-15% increase corresponding to anthocyanins or UV-A absorbing flavonoids. Treatment with visible light only gave a significant increase in quercetins from a not detectable level to 8.3 mg100 g-1 fresh matter in the peel of green
apples only. Otherwise, there were no changes in any apples due to irradiation or parallel storage in the dark.
Table 3. Effect of postharvest irradiation on various constituents and properties in the peel of green Aroma apples (mean, n = 6) Constituent or property (mg100 g-1 fresh matter if not otherwise stated) Peel Epicatechin Procyanidins (B1 + B2) Phloridzin Cyanidine-3-galactoside Quercetin-3-glycosides Sum flavonoids Chlorogenic acid Sum phenols Total phenols (mg gallic acid eq. 100 g-1 fresh) ORAC (mol Trolox eq./g fresh) L-ascorbic acid Peel + flesh Soluble solids (Brix) Titratable acidity (% malic acid eq. 100 g-1 juice) *,*** - significant at P0.05 or 0.0, respectively01 ND = not detectable Control (dark) 18.3 11.9 1.4 ND 2.5 34.1 4.4 38.5 355 3.9 1.7 8.7 0.53 After Vis + UV-B 24.9 11.9 1.5 8.9*** 48.7*** 95.9*** 18.0*** 114*** 507 8.4* 10.2*** 9.4 0.54
Using individual fruit data from all the treatments correlations were calculated. The antioxidant capacity of the peel was better correlated to the sum of phenols (r = 0.70, P < 0.001) than to the concentration of L-ascorbic acid (r = 0.54, P < 0.001). Individual phenols were positively correlated to the ORAC value (r=0.51-0.65, P < 0.001) with the lowest correlation to procyanidins (r = 0.46, P < 0.01). The red colour in the apple skin (a*) had a positive correlation to the sum of phenols (r = 0.91), L-ascorbic acid (r = 0.83), soluble solids (r = 0.66) and antioxidant capacity (r = 0.63) in the apple peel, all with P < 0.001. The total phenols (Folin-Ciocalteu) were correlated to the sum of phenols (r = 0.88, P < 0.001), but chlorogenic acid was poorly correlated to the concentration of flavonoids (r = 0.44, P < 0.01). Titratable acidity was not correlated to any of the other parameters measured. DISCUSSION The present study shows that it is possible to induce not only phenolics, as has been found by others, but also to increase other health-related constituents in fruit and vegetables by postharvest irradiation treatment. Increases in total antioxidant capacity and L-ascorbic acid were demonstrated for apples. At the same time the sensory related properties soluble solids and titratable acidity
were not changed. Nevertheless, looking at the individual variation among all apples having a history of differing pre- and postharvest conditions we found that antioxidant capacity was correlated to the sum of phenols and to ascorbic acid in the peel. However, the intensity of red skin colour correlated better to these parameters, and, in addition, but weaker, also to soluble solids. The only parameter not affected by irradiation treatment and not correlated to any of the other parameters was titratable acidity. Thus, redness would be a good sensory parameter to indicate health value and sweet taste, since the ratio soluble solids/titratable acidity increased with the intensity of red colour. Furthermore, red colour would be a better indicator than antioxidant capacity (ORAC method). This indicates that both health-related properties and flavour in Aroma apples are dependent upon preharvest radiation conditions, and that postharvest irradiation can increase the health value but not change the flavour, provided that the increase in phenolic constituents does not change the flavour. The broccoli experiment was probably carried out at sub-optimal conditions and the number of constituents analysed was rather small. The lack of significant increase in flavonols could have been caused by the large variation in contents both before and after radiation treatment, or by the presence of UVA during the treatment. UV-A has been found to counteract the flavonoid inducing effect of UV-B in another Brassica species (Wilson et al. 2001). Irradiation had only effect on the directly exposed organs, i.e. on the skin, but not the parenchyma of apples, and on the flower buds, but not the stalks of broccoli florets. The fact that controlled radiation treatment post-harvest can change the health-related quality is an indication that also the incident light during distribution of fruit and vegetables could have an effect, which might be positive or negative depending upon the conditions. CONCLUSIONS Although not fully optimised, we have shown for one fruit and one vegetable that postharvest irradiation with Vis and UV-B can increase the health value, and at the same time maintain an acceptable quality for marketing. Irradiation with other composition and intensity may have other effects and this should be investigated further.
REFERENCES Arakawa O., Hori Y., Ogata R. 1985. Relative effectiveness and interaction of ultraviolet-B, red and blue light in anthocyanin synthesis of apple fruit. Physiol. Plant. 64: 323-327. Bengtsson G.B., Schner R., Lombardo E., Schner J., Borge G.I.A., Bilger W. 2006. Chlorophyll fluorescence for non-destructive measurement of flavonoids in broccoli. Postharvest Biol. Technol. 39: 291-298. Bilger W., Johnsen T., Schreiber U. 2001. UV-excited chlorophyll fluorescence as a tool for the assessment of UV-protection by the epidermis of plants. J. Exp. Bot. 52: 2007-2014.
Cao G., Prior R.L. 1999. Measurement of oxygen radical absorbance capacity in biological samples. Methods Enzymol. 299: 50-62. Hagen S., Solhaug K.A., Bengtsson G.B., Borge G.I.A., Bilger W. 2006. Chlorophyll fluorescence as a tool for non-destructive estimation of anthocyanins and total flavonoids in apples. Postharvest Biol. Technol. [In press.] Lancaster J.E., Reay P.F., Norris J., Butler R.C. 2000. Induction of flavonoids and phenolic acids in apple by UV-B and temperature. J. Hortic. Sci. Biol. 75: 142-148. Ubi B.E., Honda C., Bessho H., Kondo S., Wada M., Kobayashi S., Morigushi T. 2006. Expression of anthocyanin biosynthesis genes in apple skin: Effect of UV-B and temperature. Plant Sci. 170: 571-578. Wilson K.E., Thompson J.E., Huner N.P.A., Greenberg B.M. 2001. Effects of ultraviolet-A exposure on ultraviolet-B-induced accumulation of specific flavonoids in Brassica napus. Photochem. Photobiol. 73: 678-684. World Cancer Research Fund, American Institute for Cancer Research 1997. Food, nutrition and the prevention of cancer: a global perspective. American Institute for Cancer Research, Washington, DC. WPYW NAPROMIENIOWYWANIA POZBIORCZEGO NA ZDROWOTNO I CECHY SENSORYCZNE JABEK I BROKUW Streszczenie Badano wpyw pozbiorczego napromieniowywania jabek i brokuw wiatem widzialnym oraz UV-A i UV-B na kilka cech organoleptycznych wiadczcych o smaku i majcych wpyw na zdrowie konsumenta. Jabka i brokuy uprawiane w Norwegii byy po zbiorze traktowane przez okres 10-12 dni rnymi dawkami wiata widzialnego i UV w warunkach kontrolowanego przechowywania w temperaturze 5o i 10oC. W owocach jabek i rach brokuu badano zawarto: kwasu L-askorbinowego, kwasu chlorogenowego i flawonoidw (met. HPLC), fenoli ogem, substancji rozpuszczalnych oraz kwasowo miareczkow. Aktywno antyoksydacyjn analizowano metod pomiaru aktywnoci absorpcyjnej rodnikw tlenkowych w ekstraktach metanolowych. Stwierdzono m.in., e w rach brokuu zawarto flawonoli i aktywno antyoksydacyjna bya duo wysza w porwnaniu z czci odygow brokuu. Zaobserwowano du zmienno naturaln pomidzy rami pochodzcymi z rnych rolin; m.in. dziesiciokrotne rnice w zawartoci flawonoli i dwukrotne rnice w aktywnoci antyoksydacyjnej. Zawarto flawonoli w rach brokuu miaa tendencj wzrostow po zastosowaniu kombinacji napromieniowywania wiatem widzialnym + UV-A+UV-B. Aktywno antyoksydacyjna brokuw nie zmienia si pod wpywem napromieniowania. Traktowanie jabek promieniowaniem UV-B i widzialnym wpyno na ich czerwony kolor, aktywno antyoksydacyjn oraz na zawartoc flawonoli, antocyjanw, kwasu chlorogenowego i askorbinowego w skrce, lecz nie miao to wpywu na poziom tych wskanikw w miszu jabek. Najwyszy efekt zaobserwowano u jabek zielonych zbieranych z wewntrznej czci korony jaboni. Badania wykazay, e zastosowanie promieniowania pozbiorczego wpywa na wzrost zawartoci specyficznych zwizkw fenolowych i innych czynnikw prozdrowotnych przy zachowaniu waciwoci sensorycznych.

CONTENT OF HIGHER FATTY ACIDS IN CHERRY STONES GROWN IN NORWAY AND SLOVENIA
Lars SEKSE1, Janez HRIBAR2, Rajko VIDRIH2 Bioforsk, Ullensvang Research Center, 5781 Lofthus, Norway 2 Biotechnicasl Faculty, Department of Food Science and Technology, Jamnikarjeva 101, 1000 Ljubljana, Slovenia
1
Summary Content of water, total fats and fatty acids composition have been determined in cherry seeds grown in Slovenia and in Norway. We used sixteen Slovenian samples of cherries: B moreau, Brooks, Burlat, Celeste, Ferrovia, Garnet, Isabella, Kordia, Lapins, Ljubljanska, New star, R12 x 10/2, Starking hardy giant, Stella, Sunburst, Vigred. Beside that we have analyzed the seed oil from five samples grown in Norway: Lapins, Celeste, Starking hardy giant, New star, Burlat. Slovenian kernels contain from 7.3 to 19.9 weight percent (w.%) of water and from 14.6 w.% to 47.2 w.% of oil. The dominant fatty acids were: linoleic acid (from 36.48 w.% to 63.47 w.%), oleic acid (from 15.89 w.% to 49.89 w.% palmitic acid (from 7.43 w.% to 14.99 w.%), stearic acid (from 2.34 w.% to 5.26 w.%), arachidic acid (from 1.12 w.% to 2.12 w.%) and linolenic acid (from 0 w.% to 1.82 w.% ). Norwegian cherry seed oil contains: linoleic acid (from 43.01 w.% to 52.52 w.%), oleic acid (from 34.26 w.% to 43.32 w.%), palmitic acid (from 8.85 w.% to 10.84 w.%), stearic acid (from 2.42 w.% to 3.46 w.%), arachidic acid (from 1.14 w.% to 1.7 w.%) and linolenic acid (from 0 w.% to 0.34 w.%). Most Norvegian cherry cultivars contain more unsaturated fatty acids compared to the same cultivars grown in Slovenia. key words: cherry stones, higher fatty acids, climatic conditions INTRODUCTION Cherry is an old commodity grown in China 4000 BC. It originates from central Asia. Hjalmarsson & Ortiz (2000) report that evidence of cherry consumption was found in graves in Norway 500 A.D. Nowadays are cherries grown all over the world. Cherries are mostly consumed as fresh fruit, some are processed in brandy, compots and marmalades. According to traditional medicine cherries have healing effect in cardiovascular and stomach diseases and they function as an diuretic. From the nutritional point of view cherries contain dietary fibers, ascorbic acid and some important minerals (Table 1). According to (Willfort 1988) cherries play some role in the development of teeth, bones and are recommended during various dietetic treatments.
Cherry stones could be an alternative source of fats. In Cherries kernel represent relatively high portion compared to the whole fruit. Cherry fats can be used as technical fats, in cosmetics and in nutrition. Cherry fats contain also phospholipids and may be an alternative source for production of phospholipids (Zlatanov & Janakieva 1998). Zlatanov & Janakieva (1998) found in cherries cv. Hebros 43.7% of fatts in dry weight. Cherry fats contain 97.7% threeacylglicerols, 0.3% free fatty acids and 1.1% phospholipids. Linolic acid is a prevalent fatty acid in cherry stones followed by oleic acid (Table 2); among saturated fatty acids cherry stones contain 15% of palmitic acid and cca. 1% of stearic acid.
Table 1. Nutrient composition of cherry mesocarp (Wills et al. 1984) Water (%) Proteins (%) Fats (%) Sugar (%) Dietary fibbers (%) Acids (%) Ascorbic acid (mg100 g-1) Content of some nutritionally important minerals K (mg100 g-1) Na (mg100 g-1) Ca (mg100 g-1) Mg (mg100 g-1) Fe (mg100 g-1) 82 0.9 0.2 13 1.5 1.1 18
240 0.3 24 9 0.3
Table 2. Content of fatty acids (%) in cherries (Zlatanov & Janakieva 1998) Palmitic acid Palmitooleic acid Stearic acid Oleic acid Miristic acid Linolic acid Linolenic acid 15.4 0.2 1.0 36.9 0.3 45.7 1.0
MATERIAL AND METHODS Fatty acid composition was analysed in cherry cultivars from Slovenia (B moreau, Brooks, Celeste, Ferrovia, Garnet, Isabella, Lapins, Ljubljanska, New star, R12x10/2, Starking hardy giant, Stella, Sunburst, Vigred and Kordia. From Norway following cultivars were included: Burlat, Celeste, Lapins, New star and Starking hardy giant. Approximately 200 kernels were used to carry out the analyses of water content, total fat content and fatty acid composition. Water content was analysed by heating the grinded seeds at 105C till constant weight. Total fat content was determined by means of Soxhlet method.
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Fatty acids were analysed according to the method described by Garces & Mancha (1993). Methyl esters of fatty acids were analysed on GC and peaks were recorded. GC: Agilent technologies 6890N. Column: SUPELCO SPB PUFA; 30m x 0.25 mm x 0.2 m. Detector: FID. Column temperature: 210C. Detector temperature: 260C. Injector temperature: 250C (split 1: 100). Carrier gas: He 1 mL/min. Injection volume: 1,0 L. RESULTS Seeds of cherry cultivars contain form 8 to 20% of water (Table 3). Cherry seeds contain from 14 to 47 % of total fats calculated on dry matter. Linolic acid is a predominant fatty acid in cherry seeds (Table 4). The average amount of linolic acid is 50%, followed by oleic acid that amounts to 35%. Among saturated fatty acids palmitic acid prevails with 10%, followed by stearic 3% and arachidic 0.5%.
Table 3. Content of water and total fats in Slovenian cherries cultivar B moreau Brooks Burlat Celeste Ferrovia Garnet Isabella Kordia Lapins Ljubljanska New star Unknown R 12 x 10/2 Starking hardy giant Stella Sunburst Vigred Water (g100 g-1) 10.9 12.4 9.98 19.9 9.4 15.7 10.2 11.2 8.05 10.5 7.3 12.1 11.7 8.8 9.3 10.3 9.5 Total fats (g100 g-1 dry matter) 26.5 30.2 17.3 33.6 15.6 33.5 36 14.6 22.8 47.2 35.8 17.6 43.9 23.4 19.4 22.5 40.1
Table 4. Content of fatty acids in total samples of Slovenian and Norvegian cherries n C 16 : 0 C 18 : 0 C 18 : 1 C 18 : 2 C 18 : 3 C 20 : 0 189 189 189 189 189 189 Average ratio (%) 10.1 3.2 35.7 49.1 0.4 0.5 Minimal ratio 7.4 2.3 15.9 36.5 0 1.12 Maximal ratio 15.0 5.3 50 63.4 1.8 2.1 Standard deviation 1.7 0.6 7.7 6.2 0.4 0.2
n total number of samples
In Slovenian samples early varieties Burlat and B moreau have the highest content of palmitic acid. Kordia has the highest content of stearic acid (Table 5). Later harvesting varieties Starking hardy giant, New star, Lapins and Sunburst have the highest content of oleic acid. On the other hand have early ripening variety Burlat the lowest content of oleic acid. Among all cultivars, burlat has the highest content of linolic acid. Later ripening varieties Starking hardy giant and Lapins have the lowest content of linolic acid.
Table 5. Fatty acid composition of cherry cultivars grown in Slovenia (as % of total fatty acids; Duncan test)
parameter cultivar B moreau Brooks Ferrovia Garnet Isabella Kordia Ljubljanska R12 x 10/2 Stella Sunburst Vigred Burlat Celeste Lapins New star Starking hardy giant C 16:0 13.53b 12.54c 10.25f 9.16h 10.24f 9.92g 9.03h 8.61i 10.01fg 10.63e 9.20h 14.71a 11.47d 9.92g 8.33i 8.53i C 18:0 4.05b 3.41f 3.53ef 3.23gh 2.47l 4.61a 2.995ij 3.03ij 3.09hi 3.88c 2.88j 2.68k 2.50l 3.47ef 3.32fg 3.71d C 18:1 24.61i 23.94i 37.56e 36.85ef 32.52h 34.35g 38.81d 40.59c 35.18g 43.55b 36.39f 16.66j 23.92i 43.96b 43.66b 48.39a C 18:2 54.87d 57.96c 46.79hi 49.09g 52.93e 49.54fg 47.44h 46.12i 50.24f 39.94l 49.89fg 62.27a 59.92b 40.82k 42.92j 37.86m C 18:3 1.34b 0.84c 0.33g 0.28ghi 0.45e 0.17lm 0.22jkl 0.19klm 0.3hij 0.39f 0.24ijk 1.74a 0.6d 0.3gh 0.23ijk 0.15m C 20:0 1.61b 1.31g 1.54bc 1.39f 1.40f 1.41ef 1.51cd 1.47de 1.22h 1.61b 1.39f 1.94a 1.6b 1.53cd 1.54bc 1.37fg
Note: Means with different superscript within groups differ significantly (P=0.05)
In cherry stones from Norway, Lapins has the highest content of palmitic acid and Burlat the lowest (Table 6). Lapins has also the highest content of oleic acid what corresponds to the situation in Slovenia. Starking hardy giant has the highest content of linolic acid what appears the opposite in Slovenia.
Table 6. Fatty acid composition of cherry cultivars grown in Norway (as % of total fatty acids; Duncan test)
Parameter cultivar Burlat Celeste Lapins New star Starking hardy giant
Note: see Table 5
C 16:0 8.96c 9.1bc 9.88a 9.33b 9.09bc
C 18:0 2.45d 3.35a 2.61c 3.14b 2.64c
C 18:1 36.34c 35.16d 42.58a 39.77b 34.9d
C 18:2 50.4b 50.7b 43.68d 46.15c 51.65a
C 18:3 0.32a 0.08c 0.06c 0.20b 0.19b
C 20:0 1.54b 1.62a 1.19d 1.42c 1.54b
Cherry seeds from Norway contain more unsaturated fatty acids (P=0.05), than seeds from the same cultivars grown in Slovenia (Table 7). Only one cultivar (New star) grown in Norway contains less unsaturated fatty acids compared to the same cultivar grown in Slovenia.
Table 7. Fatty acid composition of cherry cultivars grown in Slovenia and Norway (as % of total fatty acids; Duncan test) Ratio of fatty acids (%) Cultivar Fatty acid Slovenia Norway C 16:0 14.71 a 8.96 b C 18:0 2.68 a 2.45 b C 18:1 16.66 b 36.34 a Burlat C 18:2 62.27 a 50.4 b C 18:3 1.74 a 0.32 b C 20:0 1.94 a 1.54 b 19.32 a saturated 12.94 b unsaturated 80.67 b 87.05 a C 16:0 11.47 a 9.1 b C 18:0 2.50 b 3.35 a C 18:1 23.92 b 35.16 a Celeste C 18:2 59.92 a 50.7 b C 18:3 0.6 a 0.08 b C 20:0 1.6 a 1.62 a saturated 15.56a 14.06b unsaturated 84.44b 85.94a C 16:0 9.92 a 9.88 a C 18:0 3.47 a 2.61 b C 18:1 43.96 a 42.58 a Lapins C 18:2 40.82 b 43.68 a C 18:3 0.3 a 0.06 b C 20:0 1.53 a 1.19 b saturated 14.91a 13.67b unsaturated 85.09b 86.32a C 16:0 8.33 b 9.32 a C 18:0 3.32 a 3.14 b C 18:1 43.66 a 39.77 b New star C 18:2 42.92 b 46.15 a C 18:3 0.23 a 0.20 a C 20:0 1.54 a 1.42 b saturated 13.19 b 13.88 a unsaturated 86.81 a 86.12 b C 16:0 8.53 b 9.09 a C 18:0 3.71 a 2.64 b C 18:1 48.39 a 34.9 b Starking hardy giant C 18:2 37.86 b 51.65 a C 18:3 0.15 a 0.19 a C 20:0 1.36 b 1.54 a saturated 13.60 a 13.26 b unsaturated 86.40 b 86.74 a
Note: see Table 5
DISCUSSION Content and ratio of fatty acids is cultivar dependent what is in agreement with Lotti et al. (1971). All cultivars contain low content of linolenic acid, similar results were published by Takagi & Itabashi (1981). Cherry oil belongs to the group of oils with prevailing linolic acid (Gunstone 1994). Fatty acid composition is cultivar dependent and there are some indications that there are differences between early and late ripening cultivars. This hypothesis needs further investigations. It was reported that there are less saturated fatty acids in seeds of stone fruits grown in northern areas compared to fruits grown in southern areas. In our experiment less saturated (palmitic, stearic and arachidic acid) was found in 4 out of 5 cherry cultivars from Norway. Among unsaturated fatty acids more linoleic acid was found in 3 out of 5 cultivars from Norway compared to the same cultivars grown in Slovenia. CONCLUSIONS Linolic acid is the prevalent acid in cherry seeds followed by oleic acid. The main difference between cherries grown in Norway and Slovenia is the content of unsaturated fatty acids. More unsaturated fatty acids was found in cherry seeds from Norway. This finding corresponds to the observation of other authors that there is more unsaturated fatty acids in seeds grown in northern (colder) areas.
REFERENCES Gunstone F. 1994. Speciality oils of vegetable origin. International Food Ingredients 3: 51-54. Garces M., Mancha M. 1993. One step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Analytical Biochemistry 211: 139-143 Hjalmarsson I., Ortiz R. 2000. In situ and ex situ assessment of morphological and fruit variation in Scandinavian sweet cherry. Sci. Hort. 85(1-2): 37-49. Lotti G., Pisano G., Anelli G., Baragli S. 1971. Gli oli di semi di una serie di cultivar di ciliege. Scienza dell'Alimentazione, 16(7): 248-253. Takagi T., Itabashi Y.1981. Occurence of mixtures of geometrical isomers of conjugated octadecatrienoic acids in some seed oils: analysis by open tubular gas liquid chromatography and high performance liquid chromatography. Lipids, 16(7): 546-551. Willfort R. 1988. Zdravilne rastline in njih uporaba. Maribor, Zaloba obzorja Maribor: 507 pp. [in Sloven] Wills R.B.H., Scriven F.M., Greenfield H. 1984. Nutrient composition of stone fruit (Prunus spp.) cultivars: apricot, cherry, nectarine, peach and plum. J. Sci. Food & Agricult. 34(12): 1383-1389. Zlatanov M., Janakieva I. 1998. Phospholipid composition of some fruit-stone oils of Rosaceae species. Fett/Lipid, 100(7): 312-315.
ZAWARTO WYSZYCH KWASW TUSZCZOWYCH W PESTKACH OWOCW CZERENI UPRAWIANYCH W NORWEGII I SOWENII Streszczenie Mierzono zawarto wody, tuszczw ogem i kwasw tuszczowych w pestkach czereni. Badaniami objto szesnacie nastpujcych odmian uprawianych w Sowenii: Bigarreau moreau, Brooks, Burlat, Celeste, Ferrovia, Garnet, Isabella, Kordia, Lapins, Ljubljanska, New Star, R12 x 10/2, Starking Hardy Giant, Stella, Sunburst i Vigret Ponadto w nastpujcych piciu odmianach rosncych w Norwegii: Lapins, Celeste, Starking Hardy Giant, New Star i Burlat oznaczono tuszcze ogem i kwasy tuszczowe zawarte w pestkach owocw. Pestki z owocw uprawianych w Sowenii zawieray od 7,3 do 19,9 procent wagowych wody i od 14,6 do 47,2% tuszczw ogem. Dominujce kwasy tuszczowe to: kwas linolowy (od 36,48% do 63,47%), kwas oleinowy (od 15,89% do 49,89%), kwas palmitynowy (od 7,43% do 14,99%), kwas stearynowy (od 2,34% do 5,26%), kwas arachidowy (1,12% do 2,12%) i kwas linolenowy (od 0% do 1,82%). Pestki owocw czereni uprawianych w Norwegii zawieray kwas linolowy (od 43,01% do 52,52%), kwas oleinowy (od 34,26% do 43,32%), kwas palmitynowy (od 8,85% do 10,84%), kwas stearynowy (od 2,42% do 3,46%), kwas arachidowy (od 1,14% do 1,7%) i kwas linolenowy (od 0% do 0,34%). Wikszo norweskich czereni zawierao wicej nienasyconych kwasw tuszczowych w pestkach owocw w porwnaniu do tych samych odmian uprawianych w Sowenii.

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THE CONTENT OF PHENOLICS IN SWEET CHERRIES (PRUNUS AVIUM L.) RELATED TO CULTIVAR AND STAGE OF MATURITY
Eivind VANGDAL1, Rune SLIMESTAD2, Lars SEKSE1 1 Bioforsk Vest Ullensvang, Lofthus, Norway 2 Plantchem, Sandnes, Norway
Summary Fruit of sweet cherry cultivars grown in Norway were analysed for content of phenolic compounds. The major phenolic compounds in fruit with dark coloured juice were anthocyanins. In yellow fleshed swet cherry fruit the major phenolic compounds were the phenolic acids neochlorogenic acid and pcoumaroylquninic acid. The major anthocyanins found in sweet cherries were cyanidine 3-rutinoside and cyanidine 3-glucoside. In addition peonidine 3rutinoside and pelargonidine 3-rutinoside were detected in fruit with dark coloured juice. Large differences were found between cultivars. The cultivar with the highest content of phenolic compounds had 17 times higher content than the cultivars with lowest registered content. Annual variations were observed. However, cultivars having high contents of phenolics one year, were also among the cultivars having the highest contents in other years. During ripening the content of anthocyanins increased significantly. Hence a significant increase in total phenolic content was found. No significant changes in the contents of phenolic acids were observed during ripening. Sweet cherry fruits are susceptible to develop cuticular fractures towards optimum harvest date. Fruit with many cuticular fractures had higher contents of phenolic acids and total phenolics than fruits with few cuticular fractures. This may indicate that phenolics play a role in the natural defence of fruit against fruit rot. key words: anthocyanins, chlorogenic acid, coumarylquinic acid, cuticular fractures, phenolic acids INTRODUCTION Phenolic compounds in fruit include anthocyanins with the characteristic colours and colourless phenolic acids. They are important in many biochemical reactions in the cells in the fruit (Van Buren 1970).
The major anthocyanins in sweet cherries are cyanidine 3-rutinoside and cyanidine 3-glucoside (Van Buren 1970). In addition to the major anthocyanins Gao & Mazza (1995) found the minor anthocyanins peonidine 3-rutinoside, pelargonidine 3-rutinoside and traces of peonidine 3-glucoside. The major phenolic acids were identified as neochlorogenic acid and p-coumaroylquninic acid. Gao & Mazza (1995) studied the variation in the contents of phenolic compounds and antioxidative effect in sweet cherry cultivars. Large variations were found between cultivars and degree of ripening, and annual variation due to climatic conditions has also been reported (Gonzalves et al. 2004). Mozetic et al. (2002) reported analyses of phenolics in Slovenian sweet cherry cultivars. Large culticar differences were found. A lot of work has focused on the phenolic compounds role in defence mechanisms to protect the fruit against decay and fruit rot (Harborne 1994, De Gara et al. 2003). In the later years the important health benefit of a diet rich in fruit and vegetables has been shown (World Cancer Research Fund 1997). This health promoting effect has ben related to the content of antioxidants (Halvorsen et al. 2002). Several major phenolic compounds are strong antioxidants (Elstner et al. 1994). In the present study sweet cherries grown in Norway were analysed for content of anthocyanins and phenolic acids. In addition slightly ripe and tree ripe fruit were analysed, as well as fruit with few or many cuticular fractures. Cuticular fractures develop in the outer layers of the fruit skin in the apical end (Glenn & Poovaiah 1989, Sekse 1995). MATERIALS AND METHODS The fruits were picked in the cultivar testing orchards at Bioforsk Vest Ullensvang Research Centre in Lofthus, Western Norway. In 2001 screening analyses of fruits of 41 different cultivars and selections were performed. In the years 2002-2004 9 cultivars were included in the study. The experiments included 6 cultivars with fruit with dark coloured juice: Burlat, Chelan, Kristin, Lapins, Ulster and Vista. The following cultivars with yellow fleshed fruit were also analysed: Merton Glory, Sue and Vega. The trees were grafted on Prunus avium L. seedling rootstock F12/1 and most of them planted in 1993. Trees of some cultivars were younger, however, not less than 5 years old. Planting space was 2x4 m and the trees were vertical axis shaped. A 1 m wide stripe in the tree row was kept weed free, and the management of the orchard was made according to standard procedures in the area with fertilizers and plant protection program. The fruits were picked at optimum harvest date. In addition fruit samples of the cultivars Ulster, Van and Lapins were picked when slightly ripe. Each fruit in samples of 100 fruits from the cultivars Merton Glory, Vega and Sue were classified according to amount of cuticular fractures. Using a stereo magnifier (Carl Zeiss, Germany, magnification 16X, objective
E. VANGDAL et al. THE CONTENT OF PHENOLICS ... 171 _____________________________________________________________________________________________________
10X), the fruits were graded into five classes as described by Sekse (1995). Fruit in class 1 had no visible cuticular fractures around the stylar scar, while class 5 fruit had many distinct fractures covering large parts of the fruit surface. Within each sample of fruit the amount of cuticular fractures was calculated as average classification. A sample of approximately 20 g of fruit were weighed and extracted in 100 ml 0.5% HCl in methanol for 24h at roomtemperature and in the dark. In some selected sweet cherry cultivars 5 single fruits were exctracted to find the variations between fruits within a cultivar. The fruits were extracted as whole fruits. However, the stones were carefully removed. After the extraction periode, the fruits were crushed and the juice filtered and mixed with the extract. The pH-differential method was used (Giusti & Wrolstad 2001). 500 l of the extract was diluted in 4.5 ml of two buffers, pH 1.03 and pH 4.52. Absorption at 520 nm and 700 nm were measured, and the content of anthocyanins was expressed in terms of cyanidin3-glucoside equivalents according to the equation: A = (A520nm A700nm)pH 1.03 (A520nm - A700nm)pH 4.52 The analysis were performed according to Price & Butler (1997), Graham (1992) and Hagerman (2002). 100 L of extract were diluted with 3 mL water and 1 ml of each reagent were added. The reaction was terminated after 15 min. The results were standarised against gallic acid. Analyses of anthocyanins and phenolic acids HPLC-analysis, as described by Vangdal & Slimestad (2005), was used to identify and quantify the phenolic compounds. Neochlorogenic acid and p-coumaroylquinic acid were detected at 320 nm, whereas the anthocyanins were recorded at 520 nm. Standards were supplied by PlantChem, Norway. The programme Excel with statistical analyses package was used for statistical analyses of differences between cultivars, maturity stages, years and amount of cuticular fractures. RESULTS AND DISCUSSION The anthocyanins cyanidine 3-rutinoside and cyanidine 3-glucoside were found in all tested cultivars. In cultivars with fruits with dark coloured juice the anthocyanins peonidine 3-rutinoside and pelargonidine 3-rutinoside were also present. According to Mazza & Miniati (1993) peonidine 3-glucoside has been detected in sweet cherries. This anthocyanin was not found in fruit of the 41 sweet cherry cultivars included in this study. In yellow fleshed cultivars no other anthocyanins than cyanidine 3-rutinoside and cyaniside 3-glucoside were observed. Cyanidine 3-rutinoside was the major anthocyanin in both cultivars with coloured fruit juice and the yellow fleshed ones. It made up from 77% to 95% of the total anthocyanins in the analysed sweet cherries (Table 1). The other anthocyanins comprised up to 17% of the total anthocyanins. The content of different anthocyanins as per cent of total anthocyanin content were quite similar in the tested cultivars. Fruits of Lapins, however, had higher content of
peonidine 3-rutinoside than the other cultivars. In Lapins 17% of the total anthocyanins was peonidine 3-rutinoside. Content of this anthocyanin as per cent of total anthocyanins ranged in the other cultivars tested from 0.2 to 8%. As shown in Table 2 large differences in anthocyanin content were found between cultivars with fruit with coloured juice. Among the cultivars with fruit with dark coloured juice the highest content was 7 times the lowest registered content. In total phenolic content the highest value was 10 times the lowest. The differences were less in the yellow fleshed cultivars. The major phenolic acids detected were neochlorogenic acid and pcoumaroylquinic acid. The contents of phenolic acids found were higher in yellow fleshed cultivars than in cultivars with fruit with dark coloured juice. However, the differences observed were not significantly different. The content of anthocyanins and total phenolic compounds were significantly higher in dark coloured fruits than in yellow fleshed fruit. The most prominent change during maturation in content of phenolic compounds was a significant increase in anthocyanins towards the ripe stage (Table 3). Hence, a significant increase in total phenolics content was found as the fruits ripened. No significant difference was found in the content of phenolic acids. Comparing the contents of phenolics in different years, differences were observed (Table 4). This has been reported by Gonzalves et al. (2004). Variations between years may explain some of the differences found when comparing our results with values reported previuosly (Gao & Mazza 1995). Another reason could be different sample preparation methods (homogenization, extraction and filtration) as discussed in a previous paper (Vangdal & Slimestad 2005). Analyses of sweet cherry fruits with few or many cuticular fractures showed larger amounts of both major phenolic acids and total phenolic compounds in fruits with many fractures. The amount of cuticular fractures increases during ripening of sweet cherries. However, the content of anthocyanins was not affected by the amount of fractures (p=0.4), indicating that the fruits with few cuticular fractures were at the same stage of maturity as the fruits with many fractures. As the fracures are wounds in the cuticula and act as infection sites for pathogens (Brve et al. 2002) the increases observed in phenolic compounds may be explained as a part of the fruits defence against pathogens.
Table 1. Content of different anthocyanins as per cent of total anthocyanin content. Average and range of 6 cultivars with dark coloured juice and 3 yellow fleshed cultivars in two years Fruit with dark coloured juice (per cent of total anthocyanin content) Average Range 87 77 - 93 9 6 13 4 0 17 0.4 01 Yellow fleshed fruit (per cent of total anthocyanin content) Average Range 95 94-96 5 4-6 -
Anthocyanin Cyanidine 3-rutinoside Cyanidine 3-glucoside Peonidine 3-rutinoside Pelargonidine 3-rutinoside
Table 2. Average and range of content (mg pr 100 g FW) of anthocyanins, phenolic acids and total phenolic compounds in sweet cherries with dark coloured juice (6 cultivars) and yellow fleshed fruit (3 cultivars). Average of two years Content of anthocyanins (mg pr 100 g FW) Dark coloured sweet Yellow fleshed sweet cherries cherries Average Range Average Range 86 11 184 2 12 6 3 - 11 8 6 11 7 3 - 10 19 17 21 99 22 - 201 29 26 33
Compound Anthocyanins Neochlorogenic acid 3-p-coumarylquinic acid Total phenolic compounds
Table 3. Content (mg pr 100 g FW) of anthocyanins, phenolic acids and total phenolic compounds in sweet cherries at different stages of maturity. Average of three dark fruited cultivars and three years. Ripe Slightly ripe (optimum harvest date) -1 (mg100g FW) (mg100g-1 FW) 6 24 30 37 36 61
Compounds Anthocyanins Phenolic acids Total phenolics
P-value 0.02 0.4 0.05
Table 4. Content (mg pr 100 g FW) of total phenolic compounds in sweet cherries in two different years. Average of 6 cultivars with dark coloured juice and 3 cultivars with yellow flesh Content of total phenolic compounds (mg100g-1 FW) 2002 2004 99 32 75 30 P-value
Sweet cherry type Dark coloured juice Yellow flesh
0.02 0.5
Table 5. Content (mg pr 100 g FW) of the major phenolic acids and total phenolic compounds in sweet cherries with few or many cuticular fractures. Average of 3 cultivars with yellow flesh and three years. Compound Neo-chlorogenic acid 3-p-Coumarylquinic acid Total phenolics Few cuticular fractrures (Class 1) 7 17 26 Many cuticular fractures (Class 5) 9 21 32 P-value 0.05 0.03 0.03
CONCLUSIONS The major anthocyanins detected in sweet cherries were cyanidine 3rutinoside and cyanidine 3-glucoside. In fruits with dark coloured juice peonidine 3-rutinoside and pelargonidine 3-rutinoside were also found. The major phenolic acids were identified as neochlorogenic acid and p-coumaroylquninic acid. Large variations in content of phenolic compounds were observed due to differences between cultivars, stages of maturity and years. Fruits with many cuticular fractures had higher contents of phenolic acids and total phenolics.
REFERENCES Brve J., Skaar E., Sekse L., Meland M., Vangdal E. 2002. Rain protective covering of sweet cherry trees effects of different covering methods on fruit quality and microclimate. Horttech. 13: 143-148. De Gara L., de Pinto M.C., Tommasi F. 2003. The antioxidant systems vis-a-vis reactive species during plant-pathogen interaction. Plant. Phys. Biochem. 41: 863-870. Elstner E.F., Osswald W., Volpert, R., Schempp H. 1994. Phenolic Antioxidants. Acta Hort. 381: 304-334. Gao L., Mazza G. 1995. Characterization, Quantification, and Distribution of Anthocyanins and Colorless Phenolics in Sweet Cherries. J. Agric. Food Chem. 43: 343-346. Glenn, G. M., Poovaiah, B. W. 1989. Cuticular properties and postharvest calcium applications influence cracking of sweet cherries. J. Amer. Soc. Hort. Sci. 114: 781-788. Gonzalves B., Landbo A.K., Knudsen D., Silva A.P., Moutinho-Pereira J., Rosa E., Meyer A.S. 2004. Effect of ripeness and postharvest storage on the phenolic profiles of sweet cherries (Prunus avium L.). J. Agric. Food Chem. 52(3): 523-530. Graham A.D. 1992. Stabilization of the Prussian blue color in the determination of polyphenols. J. Agric. Food Chem. 40: 801-805. Guisti M., Wrolstad R.E. 2001. Current Protocols in Food Analytical Chemistry, unit F1.2. Hagerman A.E. 2002. The Tannin Handbook: 45-46. Halvorsen B.L., Myhrstad M.C.W., Barikmo, I., Hvattum E., Remberg, S.F., Wold A.B., Haffner K., Baugerd, H., Frost Andersen L., Moskaug J.., Jacobs D.R., Blomhoff, R. 2002. A Systematic Screening of Total Antioxidants in Dietary Plants. J. Nutr. 132: 461-471. Harborne J.B. 1994. Do natural plant phenols play a role in ecology? Acta Hort. 381: 36-43. Mazza G., Miniati E. 1993. Anthocyanins in Fruits, Vegetables, and Grains. CRC Press, Boca Raton, USA, 362p. p:57. Mozetic B., Trebse P., Hribar J. 2002. Determination and quantification of anthocyanins and hydroxycinnamic acids in different cultivars of sweet cherries (Prunus avium L.) from Nova Gorica region (Slovenia). Food Tech. and Biotech. 40(3): 207-212. Price M.L., Butler L.G. 1977. Rapid visual estimation and spectrophotometric determination of tannin content of sorghum grain. J. Agr. Food Chem. 25: 1268-1273. Sekse L. 1995. Cuticular fracturing in fruits of sweet cherry (Prunus avium L.) resulting from changing soil water contents. J. Hort. Sci. 70: 631-635.
Van Buren J. 1970. Frit phenolics. 269-304. In: The biochemistry of fruits and their products Volume 1 (A.C. Hulme ed.) Academic Press, London, UK. Vangdal E., Slimestad R. 2005. Methods to determine antioxidative capacity in fruit. J. Fruit Ornam. Plant Res. 13: (in press). Vangdal E., Slimestad R., Sekse L. 2006. Phenolics and other compounds with antioxidative effect in stone fruit Preliminary results. Presentation at The 8. International Plum and Prune Symposium, Lofthus, Norway 2004 (accepted for publication in Acta Hort.) World Cancer Research Fund / American Institute for Cancer Research 1997. Food, Nutrition and the Prevention of Cancer: A Global Perspective. American Institute of Cancer Research, Washington DC.
ZAWARTO FENOLI W CZERENIACH (PRUNUS AVIUM L.) W ZALENOCI OD ODMIANY I STADIUM DOJRZAOCI Owoce czereni odmian uprawianych w Norwegii analizowano pod wzgldem zawartoci zwizkw fenolowych. Gwnymi zwizkami fenolowymi w owocach o ciemno zabarwionym soku byy antocyjany, natomiast w owocach o jasnym miszu wystpoway gwnie kwasy fenolowe, kwas neochlorogenowy i kwas pkumarylochinowy. Gwnymi antocyjanami obecnymi w czereniach byy 3-rutynozyd cyjanidyny i 3-glukozyd cyjanidyny. Dodatkowo w owocach o ciemnym soku wykryto 3-rutynozyd peonidyny i 3-rutynozyd pelargonidyny. Obserwowano due rnice w zawartoci zwizkw fenolowych pomidzy odmianami. Odmiana o najwyszej zawartoci zwizkw fenolowych miaa 17 razy wysz ich zawarto ni odmiany, u ktrych zanotowano najnisz zawarto zwizkw fenolowych. Obserwowano rwnie zmiany w poszczeglnych latach bada. Jakkolwiek odmiany, ktre miay wysok zawartoci fenoli w jednym roku byy rwnie obecne wrd odmian o najwyszej zawartoci fenoli w innych latach. Podczas dojrzewania zawarto antocyjanw znacznie si zwikszaa. Wskutek tego nastpowa znaczny wzrost oglnej zawartoci fenoli. Nie obserwowano natomiast podczas dojrzewania istotnych zmian w zawartoci kwasw fenolowych. Owoce czereni w czasie optymalnym do zbioru s podatne na pknicia skrki. Owoce z wieloma pkniciami skrki miay wysz zawarto kwasw fenolowych i oglnych fenoli ni owoce z niewieloma uszkodzeniami. Moe to wskazywa na rol fenoli w naturalnej obronie owocw przeciwko gniciu.

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THE EFFECT OF VINEYARD TREATMENTS TO INCREASE BERRY SIZE ON CHEMICAL COMPOSITION AND FRUIT QUALITY IN SEEDLESS TABLE GRAPES
Susan LURIE, Amnon LICHTER, Yakob VINEKUR, Victor RODOV Department of Postharvest Science, Volcani Center, Agricultural Research Organization, Bet Dagon 50250, Israel
Summary There are several vineyard management techniques which can be used to increase the size of seedless table grapes, including spraying with gibberellic acid (GA) and girdling. Spraying with GA increases grape size by increasing intracellular water content. Girdling works by diverting sugars and other photoassimilates to the berries. Spraying with GA and girdling can be used together to produce large grapes with a high sugar content. However, these techniques can also alter the content of other taste components, including polyphenols, which give grapes an unpleasant astringent taste. The aim of this study was to determine how spraying with GA and early or late girdling, alone or in combination, affect size, soluble solids content (SSC), titratable acidity (TA), total phenolics content, tannin content, and antioxidant activity in Superior seedless table grapes, and to correlate the results with taste testing scores for sweetness, sourness and astringency. The taste testers clearly preferred the unsprayed late-girdled grapes, giving them the highest scores for sweetness and the lowest scores for sourness. These grapes had the highest SSC and the lowest TA. The testers clearly disliked the sprayed early-girdled grapes, giving them the lowest scores for sweetness and the highest scores for sourness. These grapes had the lowest SSC and the highest TA. The testers assigned the highest scores for astringency to the sprayed early-girdled grapes, which had a total phenolics content more than twice that of unsprayed, ungirdled grapes. key words: Table grapes, gibberelin acid (GA), girdling, chemical composition, polyphenols, antioxidant activity INTRODUCTION The main taste component of seedless table grapes is sugar (Sonego et al. 2002). Therefore, the main harvest criterion for table grapes is soluble solids content. However, there are many other compounds which contribute to the taste of particular varieties, including phenolic compounds such as polyphenols, which can give grapes an unpleasant astringent taste. Procyanidins are a class of
polyphenols consisting of chains of the flavan-3-ol monomers catechin and epicatechin. Procyanidins react with saliva and oral mucus to create the sensation of astringency (Lea 1992). As grapes ripen, astringency decreases as polyphenols either breakdown into monomers or aggregate into tannin compounds, which do not have a significant effect on taste (Brossaud et al. 2001). However, in some years, the astringent taste does not entirely disappear by harvest time, prompting consumer complaints. There are several vineyard management techniques which can be used to increase the size of seedless table grapes. These include irrigation, fertilization, hand thinning the clusters, girdling the grapevine, and applying the plant hormone gibberellic acid (GA). GA decreases fruit set when applied at blossoming time, and increases berry size when applied later. Treatments to increase berry size may affect the rate at which astringency decreases during the ripening process. In this study, we compare the effects of girdling and spraying with GA on astringency in Superior table grapes. MATERIALS AND METHODS The experiment was conducted over a two-year period in vineyards in Moshav Lachish in central Israel. In the first year, grapes were harvested from two vineyards when they met the criteria for commercial ripeness. At one vineyard, the vines were sprayed with GA, but left ungirdled. At the other vineyard, half the grapevines were sprayed with GA and girdled, and the other half was neither sprayed nor girdled, and served as the control. After harvest, the grapes were analyzed in terms of maturity indices. The grapes were then peeled, and the peels and flesh were frozen separately for later analysis. In the second year, four rows of grapevines in one vineyard were divided into random replicates of six plants each. Half of the plants were sprayed with 15 g/l GA eight weeks before harvest. The other half were left unsprayed. Girdling was performed on vines of both sprayed and unsprayed plants either early (eight weeks before harvest) or late (two weeks before harvest). Some sprayed plants and unsprayed plants were left ungirdled. After harvest, the grapes were analyzed in terms of maturity indices. Taste testing was carried out. The grapes were then peeled, and the peels and flesh were frozen separately for later analysis. Maturity indices were measured in five bunches of grapes from each replicate for each treatment. Twenty grapes from each bunch were weighed. The grapes were then juiced before further analysis. Soluble solids content (SSC) was measured with a refractometer. Titratable acidity (TA) was measured by titrating against 0.1 N NaOH, and was recorded as equivalents of tartaric acid. Taste testing was performed by two panels, the first consisting of twenty untrained testers, and the other consisting of six trained testers. Parameters recorded included astringency, sweetness and sourness. All were scored on a scale from 1 to 10.
S. LURIE et al. THE EFFECT OF VINEYARD TREATMENTS ... 179 _____________________________________________________________________________________________________
In order to determine total phenolics content, the grapes were first lyophilized and ground in a glass homogenizer. The powder was extracted three times with 50% methanol for a final volume of 5.5 ml. Aliquots were analyzed for total phenolic compounds using the FolinCiocalteau reagent. Tannins were determined by precipitating the tannins in 2 ml of extract with 100 mg polyvinyl polypyrrolidone (PVPP), centrifuging, measuring the remaining phenolic compounds in the supernatant, and subtracting this amount from the total phenolic compounds. Gallic acid was used as the standard. Another aliquot was analyzed for antioxidant activity by measuring the scavenging capacity of the extract for the 2,2-azino-bis-(3-ethylbenzothiazoline)6-sulfonate cation radical (ABTS+). 1 mM Trolox was used as the standard (Vinekur et al. 2006). All data were elaborated by analysis of variance, followed by means separation using the Student-Newman-Keuls test at P 0.05. All calculations were performed with the help of the SPSS software package. RESULTS AND DISCUSSION The results from the first year are presented in Table 1. Maturity indices at harvest time were about the same for all three treatments. The control grapes were much smaller than the sprayed, ungirdled grapes. Total phenolics content per unit of fresh weight was higher in the peels than in the flesh. Antioxidant activity was also higher in the peels than in the flesh, although there was not always a strict relationship between total phenolics content and antioxidant activity. Total phenolics content and antioxidant activity were twice as high in the sprayed, girdled grapes than in either the sprayed, ungirdled grapes or the control grapes. The results from the second year are presented in Table 2. Soluble solids content was lower in sprayed grapes than in unsprayed early-girdled grapes or the unsprayed late-girdled grapes. This was because spraying with GA increased the size of the grapes by increasing water content, thereby diluting the sugar content. Titratable acidity was not affected by either spraying with GA or girdling. Because of the number of treatments, taste testing with the untrained panel was conducted twice: once in the morning, and once in the afternoon. The treatments were divided between the two sessions. Sprayed ungirdled grapes and unsprayed ungirdled grapes were included at both sessions. The results of taste testing with the untrained panel are presented in Table 3. The scores for sweetness given by the untrained testers were highly correlated with SSC. They clearly preferred the unsprayed late-girdled grapes, giving them the highest scores for sweetness and the lowest scores for sourness. Chemical analysis revealed that these grapes had by far the highest SSC and the lowest TA. The panel clearly disliked the sprayed early-girdled grapes, giving them the lowest scores for sweetness and the highest scores for sourness. These
grapes had the lowest SSC and the highest TA. The panel did not find any clear differences in sweetness and sourness among the other treatments.
Table 1. Ripeness parameters, total phenols and antioxidants of 'Superior' table grapes measured at harvest from the first season
Girdling GA + + + Peel Pulp Peel Pulp phenols phenols antiox. antiox. (%) (%) (g) (GE/g FW) (TE/g FW) 15.70.9ab 0.620.02a 9.70.8a 4.10.9a 0.90.3a 75.57.6a 123.5a 16.71.6a 0.520.02c 4.90.3c 1.90.6b 0.20.1b 52.76.5b 1.90.5b 14.70.9b 0.570.03b 7.40.7b 1.30.2b 0.40.2b 35.72.8c 2.20.1b SSC TA Weight
GE gallic acid equivalents; TE Trolex equivalents Table 2. Ripenenss parameters of 'Superior' table grapes measured at harvest from the second season GA + + + Girdling Early Late + + + + SSC (%) 15.21.23b 15.90.67b 15.450.73b 17.20.28a 17.30.55a 15.30.56b TA (%) 0.570.05a 0.520.07ab 0.530.05ab 0.530.02ab 0.480.04b 0.530.05ab Weight (g) 6.240.32a 6.301.17ab 6.530.49ab 5.520.43b 5.890.92ab 5.580.49ab
Table 3. Taste tests of 'Superior' grapes after harvest from the second season. Half the grape treatments were taste tested in the morning and half in the afternoon with two treatments, with and without GA with no girdling, being tasted at both times. There were 20 tasters. Rating was from 1 to 10 GA + + + Girdling Early Late + + + + Sweetness Morning Afternoon 5.77 6.95 6.50 6.70 6.90 7.36 6.68 6.90 Acidity Morning Afternoon 5.00 4.55 4.50 4.00 4.30 3.50 3.86 4.36
Table 4. Astringency rating (on a scale from 1 to 10), total phenols, tannins and total antioxidants in grapes from the second season that received a GA spray and early girdling, or no GA or girdling GA + Girdling Early + Astringency rating 1st test 2nd test 3.42 4.57 1.75 2.37 Total phenols Tannins (GE/g FW) 7.40.5a 3.60.3a 3.20.2b 1.70.2b Antioxidants (TE/g FW) 6.50.3a 2.70.1b
GE gallic acid equivalents; TE Trolex equivalents
The untrained testers also could not find any clear differences in astringency among the treatments. Scores for astringency ranged from 1.76 to 2.95. Taste testing with the trained panel was performed on two different days. The results of taste testing with the trained panel are presented in Table 4. The trained testers assigned the highest scores for astringency to the sprayed early-girdled grapes. They gave these grapes a score of 3.42 on the first day, and 4.57 on the second day. The testers assigned the lowest scores for astringency to unsprayed ungirdled grapes. They gave these grapes a score of 1.75 on the first day, and 2.37 on the second day. Total phenolic compounds, tannins and antioxidant activity were measured in both groups of grapes. Total phenolic compounds and tannins were twice as high in the most astringent grapes than in the least astringent grapes. Antioxidant activity was also significantly higher in the most astringent grapes than in the least astringent grapes. Spraying with GA and girdling had an effect on maturity indices in both years of the experiment. Fruit size was greatest in the sprayed early-girdled grapes. Early girdling directs more of the photo-assimilates to the berries, increasing turgor and causing cell expansion (Peacock et al. 1977). GA also causes cell expansion (Guelfat-Reich & Safran 1973, Weaver & McCune 1959). The grapes sprayed with GA were larger, but had a lower SSC because the sugar inside them was diluted with water. The sprayed grapes also ripened slower, as could be seen by their higher TA content at harvest time. If the sprayed grapes had been left on the vine to ripen longer, SSC and TA would have eventually reached the same levels as in the unsprayed grapes. However, because all the grapes in this study were harvested on the same day, the sprayed grapes were significantly less ripe than the unsprayed grapes. In the second season, SSC was highest in the unsprayed early-girdled grapes and the unsprayed late-girdled grapes. This was consistent with our expectations. Harvest criteria for table grapes include color, size, SSC, TA, and SSC/TA ratio. The criterion best correlated with the results from the taste testing was SSC, although this accounted for only 60% of the variability (Sonego et al. 2002). In this study, the grapes which received the best scores in the taste testing were the unsprayed late-girdled grapes, which had the highest SSC and the lowest TA. Little research has been done on astringency in table grapes. On the other hand, polyphenols in wine grapes have been thoroughly studied, not only because they impart unpleasant astringent and bitter tastes to the wine, but also because they play a role in the oxidation and browning processes (Brossaud et al. 2001, De Freitas & Glories 1999). Procyanidins in wine are potent scavengers of free radicals, and also help protect against arteriosclerosis (Ricardo da Silva et al. 1991, Rigo et al. 2000). The concentration and composition of procyanidins in grapes depend on the location of the vineyard, weather conditions during the growing season, and the specific cultivar under cultivation. In this study, the content of phenolic compounds was significantly affected by two vineyard management techniques:
spraying with GA, and girdling. Further study is needed to determine how GA and girdling affect the content of phenolic compounds in table grapes. In this study, the grapes with a higher total phenolics content had an unpleasant astringent taste. Paradoxically, they also had a higher antioxidant activity, and were therefore more beneficial for human health. However, phenolic compounds are not the only substances which contribute to antioxidant activity. Other compounds, especially ascorbic acid, also play a role. In some grape cultivars, ascorbic acid is in fact the greatest component of antioxidant activity (Vinson et al. 2001). Therefore, it should be possible to decrease the content of phenolic compounds in grapes without reducing antioxidant activity. Further research is needed to develop techniques for improving fruit quality and taste in Superior table grapes while reducing the content of phenolic compounds responsible for astringency.
REFERENCES Brossaud F., Cheynier V., Noble A.C. 2001. Bitterness and astringency of grape and wine polyphenols. Aust. J. Grape and Wine Research, 7:33-39. De Freitas V., Glories Y. 1999. Concentration and compositional changes of procyanidins in grape seeds and skin of white Vitis vinifera varieties. J. Sci. Food Agric. 79: 1601-1606. Guelfat-Reich S., Safran B. 1973. Maturity responses of Sultanina grapes to gibberellin acid treatments. Vitis 12: 33-37. Lea A.G. 1992. Flavour, colour, and stability in fruit products: the effect of polyphenols. In: Plant Polyphenols (ed. R.W. Hemingway and P.E. Laks) Plenum Press, NY, pp. 827-837. Peackock W.L., Jensen F., Else J., Leavitt G. 1977. The effects of girdling and ethephon treatments on fruit characteristics. Amer. J. Enol. Vitic. 28: 228-231. Ricardo da Silva J.M., Darmon N., Fernandez Y., Mijavila S. 1991. Oxygen free radical scavenger cipacity in aqueous models of different procyanidins from grape seeds. J. Sci. Food Chem. 39: 1549-1552. Rigo A., Vianello F., Cleminti G., Rosetto M. Scarpa M. Vrhovsek U., Mattivi F. 2000. Contribution of the proanthocyanidins to the peroxy-radical scavenging capacity of some Italian red wines. J. Agric. Food Shcem. 48: 1996-2002. Sonego L., Zuthi Y., Kaplonov T., Kosto I., Lurie S. 2002. Factors affecting taste scores of early season seedless table grapes Mystery and Prime. J. Agric. Food Chem., 50: 121-125. Vinekur Y., Rodov V., Reznick N., Goldman G., Horev B., Umiel N., Friedman H. 2006. Rose petal tea as an antioxidant-rich beverage: cultivar effects. J. Fd Sci. 71:S42-S47. Vinson J.A., Su X., Zubik L., Bose P. 2001. Pehnol antioxidant quantity and quality in foods: fruits. J. Agric. Food Chem. 49: 5315-5321. Weaver R.J., McCune S.B. 1959. Effect of gibberllin on seedless Vitis vinifera. Hilgardia 29: 247-275.
WPYW SPOSOBU TRAKTOWANIA WINOROLI NA ZWIKSZENIE WIELKOCI, SKAD CHEMICZNY I JAKO BEZPESTKOWYCH OWOCW WINOGRON Streszczenie Jest wiele technik prowadzenia rolin winoroli, ktre pozwalaj na zwikszenie wielkoci bezpestkowych owocw m.in. opryskiwanie kwasem giberelinowym (GA) i obrczkowanie. Opryskiwanie GA powoduje wzrost owocw winogron poprzez zwikszenie zawartoci wody w przestrzeniach midzykomrkowych. Obrczkowanie natomiast powoduje przepyw cukrw i innych foto-asymilantw do owocw. Opryskiwanie winoroli GA i obrczkowanie moe by stosowane jednoczenie, co pozwala na zwikszenie wielkoci owocw i zawartoci cukrw. Zabieg taki moe jednak prowadzi do zmiany zawartoci skadnikw odpowiedzialnych za smak, wczajc polifenole, co daje nieprzyjemny cierpki smak owocw. Celem bada byo okrelenie wpywu opryskiwania GA i wczeniejszego lub pniejszego obrczkowania, stosowanego samodzielnie lub z opryskiwaniem, na wielko owocw, zawarto substancji rozpuszczalnych (SSC), kwasowo miareczkow (TA), ogln zawarto fenoli, tanin i aktywno antyredukcyjn bezpestkowych winogron odmiany Superior i korelacj tych rezultatw ze smakiem, a szczeglnie sodkoci, kwanoci i cierpkoci owocw. Oceniajcy zdecydowanie preferowali owoce z winoroli nie opryskiwanych i pniej obrczkowanych dajc im najwysz ocen pod wzgldem sodkoci i najnisz pod wzgldem kwanoci. Owoce te zawieray najwicej substancji rozpuszczalnych (SSC) i miay najnisz kwasowo miareczkow (TA). Oceniajcy zdecydowanie negatywnie ocenili owoce z winoroli opryskiwanych GA dajc im najnisz ocen pod wzgldem sodkoci i najwysz pod wzgldem kwasowoci. Owoce te miay najnisz zawarto substancji rozpuszczalnych (SSC) i najwysza kwasowo miareczkow (TA). Najwysz ocen pod wzgldem cierpkoci uzyskay owoce z winoroli opryskiwanych GA i obrczkowanych we wczeniejszym terminie, ktre zawieray dwukrotnie wicej substancji fenolowych ni owoce pochodzce z rolin nie opryskiwanych i nie obrczkowanych.

A COMPARISON OF THE QUALITY OF WELL KNOWN AND SCAB RESISTANT APPLES IN EXPERT AND CONSUMER EVALUATION
Dorota KONOPACKA1, Katarzyna. JESIONKOWSKA1, Krzysztof RUTKOWSKI1, Witold POCHARSKI1, Kazimierz TOMALA2 1 Department of Storage and Processing, Research Institute of Pomology and Floriculture Pomologiczna 18, 96-100 Skierniewice, Poland 2 Department of Pomology, Warsaw Agricultural University, Nowoursynowska 166, 02-787 Warsaw, Poland
Summary The objective of this study was to compare the quality of well-known (standard cultivars) and scab resistant apples. Out of 12 standard and 5 scab resistant cultivars tested by experts, 3 standard and 3 scab resistant ones which were characterized by a high quality assessment and different specific traits described by PCA analysis were chosen for consumer tests. The chosen cultivars included Lodel, Melrose and Spartan (the standard) and Rajka, Rubinola and Topaz (scab resistant). Consumer tests were performed on adult citizens of Skierniewice in 2004 and 2005. The appearance and taste of apples were evaluated according to hedonic scale. The appearance of scab resistant cultivars seemed to be more sensitive to seasonal variability than the standard ones. In 2004 the scab resistant cultivars were perceived as less attractive than the standard ones, whereas in 2005 their appearance was more attractive and the differences between both groups were not significant. In the case of taste attribute in both seasons scab resistant cultivars were usually assessed lower than the standard ones, particularly in comparison to Melrose. Other standard cultivars were assessed usually between 5 and 6 points in 1-9 scale. Scab-resistant Rubinola was perceived on the same level as the well-known Spartan. The lowest taste appreciation was scored by Topaz (scab resistant) and Lodel (well-known cv.). The expert assessment indicates that a high firmness, juiciness and flavour of ripe apple increase the consumer acceptance of the apple taste. It was also found that the consumer evaluation is significantly correlated with the expert assessment (r = 0.72). key words: apple, sensory assessment, quality attributes, taste, appearance INTRODUCTION As the market of apples is more and more saturated, producers and managers are forced to search for ways of making consumers interested in their prod-
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ucts in order to be competitive. Scab resistant cultivars give growers the opportunity of reducing the cost of disease control, lessening the risk of environmental contamination associated with fungicide applications, and meeting consumer demands for reduced pesticide residue on the produce (Kuhn & Thybo, 2001). Especially the last issue needs attention because the care for food safety and human health is increasingly important for consumers (Luning et al. 2002). According to surveys performed by Baker (1999) and by Baker & Crosbie (1994) the consumer segment represented by people mostly interested in the safety issues increased from 16 to 49% during five years. Although in recent years several disease resistant varieties have been released which require only a very restricted use of fungicides, such cultivars are not popular since their fruits in most cases do not meet European consumer and grower preferences. Nevertheless tremendous breeder efforts have led to introducing new scab resistant cultivars which combine good resistance with improved taste and storability properties. To increase the acceptance of disease resistant apples, their diffusion and commercial success, marketing strategies should be based on a good knowledge of consumers quality orientation and quality perception (Steenkamp & Van Trijp 1996). Knowledge about sensory quality can facilitate the choice among many scab resistant cultivars for both fruit grower and the consumer (Kuhn & Thybo 2001). The aim of the current study was to compare the sensory quality evaluation of standard and scab resistant apples done both by expert panel and adult consumers. MATERIALS AND METHODS Twelve well-known cultivars (Alwa, Golden Delicious, Idared, Spartan, Gala, Jonagold, Lodel, Szampion, Gloster, Holiday, Melrose, Rubin) and five scab resistant ones (Rajka, Topaz, Rubinola, Wars, Sawa) were chosen as experimental material. Apples were harvested at optimal ripening stage and stored in normal atmosphere for about 2 months. After storage apples were kept for 24 hours at ambient temperature and then subjected to instrumental measurements and sensory assessment. The consumer tests were performed on apples stored in the cold room two weeks longer than those used for expert panel assessment. In this case apples were removed from cold storage also 24 hours prior to evaluation date. The investigation was performed in two successive seasons, 2004 and 2005. To check the objective parameters of apple quality the following analysis were performed: firmness by puncture test using Instron Model 4303 equipped with Magness-Taylor probe (N), juiciness according to the procedure described by Konopacka & Pocharski (2001) (%), titratable acidity (% of malic acid) and soluble solid content by refractometric method (%). The objective parameters were measured on the date of expert panel assessment and are presented in Table 1.
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Table 1. Quality characteristics of apples after 2 months of cold storage, cultivars used for consumer tests are marked in bold. Firmness (N) 2004 2005 61.5 68.1 72.7 64.5 87.8 65.6 39.5 63.3 61.1 45.8 77.3 69.2 49.3 53.5 43.3 44.5 71.3 74.1 62.7 65.7 38.5 37.1 53.1 53.9 43.9 55.7 49.3 48.1 32.8 32.1 60.7 79.7 36.2 35.7 Juiciness (%) 2004 2005 25.8 27.0 27.6 30.9 36.3 30.5 28.4 22.8 35.1 27.6 32.6 33.3 29.9 33.5 29.1 26.9 30.2 39.9 29.0 23.9 30.4 26.3 27.6 31.4 29.8 22.8 28.0 14.8 26.7 23.7 31.5 32.0 30.3 19.1 Soluble solids (%) 2004 2005 14.4 13.9 12.5 12.4 14.3 15.5 11.8 15.9 14.0 14.1 13.0 12.0 13.5 13.6 13.9 14.1 14.9 14.2 13.8 13.8 11.8 13.3 13.3 12.7 14.0 14.3 13.9 14.6 13.5 12.6 14.1 14.7 13.9 15.0 Acidity (%) 2004 2005 0.455 0.669 0.365 0.306 0.495 0.618 0.362 0.584 0.529 0.476 0.656 0.610 0.456 0.423 0.522 0.523 0.539 0.496 0.393 0.457 0.289 0.397 0.476 0.575 0.575 0.614 0.526 0.585 0.563 0.723 0.744 0.863 0.379 0.535
Cultivars Alwa Gala Gloster Golden D Holiday Idared Jonagold Lodel Melrose Spartan Szampion Rubin Rajka Rubinola Sawa Topaz Wars
The expert panel consisted of 12 people recruited from the staff of Research Institute of Pomology and Floriculture, trained and experienced in performing sensory assessment of apples. During three successive days experts assessed 17 cultivars. Each day 7 cultivars were evaluated (3 or 4 standard, 1 or 2 scab resistant with Topaz and Idared as the reference cultivars) in two repetitions (one session took place in the morning and another in the afternoon). Apples were cut into 8 segments. Two opposite segments (with skin but without core) were served as an individual sample. Each sample was served in a covered individual container to avoid aroma loss or aroma cross contamination. During one session seven samples were presented randomly to the experts. The evaluation was performed in separate booths, between samples experts used non-carboned mineral water as a neutralizer. The experts assessed 13 qualitative apple traits, which had been signed with consecutive numerals: acidic aroma (1), aroma of ripe apples (2), aroma of other fruit (4), cut grass aroma (4), other aroma (5), firmness (6), juiciness (7), sweetness (8), sourness (9), astringency (10), flavour of ripe apple (11), other flavours (12) and overall quality (13). Each attribute was rated on a continuous linear scale with anchor points at each end marked as 0 and 100 points. The scales were designated with accurate definitions and terms. The obtained results were transposed to numeric values con-
Scab resistant
Well-known
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sidering the whole scale to be 10 subjective units. The data were collected through the computerized data collecting system ANALSENT NT (LABNT), developed at Polish Academy of Sciences, Warsaw, Poland. The consumer survey was performed in the first week of December in 2004 and 2005 on adults respondents, mainly citizens of Skierniewice (a town of about 50 000 inhabitants). The number of respondents was 99 and 120 respectively in 2004 and 2005. Consumers were asked to assess the appearance and taste of 6 cultivars, 3 well-known (Lodel, Melrose and Spartan) and 3 scab resistant (Rajka, Rubinola and Topaz). Both appearance and taste assessments were performed using 1 to 9 hedonic scales. For the external assessment 4-5 apples of the same cultivar were presented on a white plate, while apples assigned for taste evaluation were cut with skin and without core into 8 segments just before being served to avoid browning effects. Cultivars were coded and presented in alternately sequences, well known and scab-resistant ones. The sensory profiling data were analyzed by the multivariate Principal Component Analysis (PCA) on standardized mean data using statistical module of ANALSENT NT (LABNT) system, developed by Polish Academy of Sciences, Warsaw, Poland. Statistical differences between consumer evaluation data were determined by the analysis of variance and Duncans Multiple Range test at 5% significance level (STATISTICA Version 7.1). RESULTS AND DISCUSSION The results of the profile sensory analyses obtained for all 17 cultivars in 2004 are presented in Fig. 1. Multivariate analysis of expert sensory data indicated the most important apple traits with different sensory quality of the investigated cultivars. The variation was generated mainly by texture attributes such as firmness and juiciness, which were placed along the horizontal axis (PC1 62.6 %). Although the traits which influenced flavour such as sourness and astringency were located along the vertical axis (PC2) explaining only 16.6%, the sourness vector was one of the longest, what means that this attribute significantly differentiated the cultivars quality evaluation. Other examined attributes such as sweetness, acidic aroma, other fruit aroma, cut grass aroma, other aroma and flavour did not substantially differentiate the quality of investigated cultivars; their vectors were very short and placed near the middle of the co-ordinate space created by PC1 and PC2. In Fig. 1 they were marked with hollow circles. After two months of storage the examined well-known cultivars were characterized with highly diversified traits and were scattered over the quality space. Gloster, Gala, Melrose and Spartan apples were characterized as the most firm and juicy cultivars. Idared was perceived as firm and sour while Lodel as sour and rather soft. Alwa, Golden Delicious and Szampion were distinctly softer than others. Spartan, Melrose and Holiday were highly scored for the overall quality. Among the scab resistant cultivars 3 of them: Wars,
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Sawa and Rajka were perceived by assessors rather as soft, with a low overall quality and in particular low juiciness. Rubinola was assessed as more firm and juicy with a higher flavour of ripe apple that gave it high overall quality. In turn Topaz was perceived differently than other scab resistant cultivars. Although it was the sourest cultivar it simultaneously was characterized by high firmness and juiciness, and thanks to that it scored highly on the overall quality. Looking for cultivars characterized by a high overall quality and simultaneously with different specific traits which would possibly cover the whole quality space two triangles were marked in the Fig. 1, which indicate two sets of cultivars. There were Melrose, Spartan and Lodel (standard cultivars) and Rubinola, Rajka, Topaz (scab resistant ones), which were then subjected to further analyses, i.e. for consumer tests.
Fig. 1. PCA analysis of well-known ( ) and scab resistant ( ) apples. Attributes which differentiated the evaluated cultivars: aroma of ripe apples (2), firmness (6), juiciness (7), sourness (9), astringency (10), flavour of ripe apple (11) and overall quality (13). Length of vectors joining attribute points with mid of co-ordinate system indicates significance of particular trait in quality differentiation. Data for 2004.
The results of consumer assessment are shown in Figures 2 & 3. As the appearance is concerned in 2004 the well known cultivars were evaluated as more attractive than the scab resistant ones (Table 2). In the next year the difference between those two categories were not significant. The reason was that in 2005 Rajka and Topaz were evaluated higher than in 2004 (Fig. 2). On the basis of the two seasons, it may be anticipated that the well-known cultivars are more stable and their appearance is not influenced by seasonal variability.
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9 Season 2004 Season 2005 bars mean 0.95 confidence intervals a a 7 b b bc b bc cd d 5 b bc
Like extremely
Like moderately
Appearance
Neither like nor dislike
Dislike moderately
1 Melrose Spartan Lodel Rajka Topaz Rubinola Well known cultivars Scab resistant cultivars
Dislike extremely
Fig. 2. Comparison of appearance of well known and scab resistant apples means marked with the same letters do not differ significantly at P=0.05 Table. 2. Comparison of appearance and taste of well-known and scab resistant apple cultivars evaluated in consumer tests, hedonic scale: 1 - dislike extremely, 9- like extremely Cultivar characteristics Well known Scab resistant Appearance 2004 2005 6.39 a 6.14 a 5.78 b 6.08 ab Taste 2004 6.05 a 5.58 b 2005 6.03 a 5.64 b
Note: means marked with the same letter do not differ significantly at P=0.05 (Duncans multiple range t-test)
In the case of taste attribute, generally in both seasons, scab resistant cultivars were assessed lower than the well-known ones. The reason was that in both seasons Melrose was indicated by consumers as the tastiest cultivar. The taste of other cultivars was assessed usually between 5 and 6 points (Fig. 3). Scab resistant Rubinola was perceived on the same level as well-known Spartan. The lowest taste appreciation was found for Topaz (scab resistant) and Lodel (well known cv.). In addition, within a particular cultivar, no significant differences in taste appreciation between years were observed, which means that consumer preferences are well-grounded. Looking for traits decisive in cultivar
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taste appreciation, based on expert sensory evaluation, the PCA projection of differences and similarities in apple quality was performed (Fig. 4). In this analysis only the data concerning cultivars also assessed by consumers were taken into account. The results are shown in the space created by the first two main components PC1 (63.1%) and PC2 (18.6%), which explains 81.8 % of the total variation in the sensory assessment of the cultivars examined in the successive seasons. For this set of cultivars the variation was explained by the texture (firmness, juiciness) as well as taste features (sweetness, sourness). Also the aroma and flavour of ripe apples and astringency differentiated the quality of investigated samples. Sensory traits characterizing the particular cultivar in the successive years were generally similar and the cultivar points for 2004 and 2005 were placed in the quality space relatively close together. The larger differences were observed for Rubinola and Lodel, which in 2005 were judged as softer and less juicy. Admittedly, this was reflected in consumer assessments but the differences were not significant (Fig. 3). In both seasons Melrose was perceived as the most firm and juicy, well balanced between sweetness and sourness and with sensible flavour of ripe apple. These features seem to correspond with high consumer appreciation. Comparing the data in Fig. 3 & 4 it may also be concluded that the combination of sourness and lack of firmness is much worse than the sweetness and lack of firmness; probably due to the fact that sweetness is usually associated with flavour of ripe apple, while sourness with astringency.
9 Season 2004 Season 2005 bars mean 0.95 confidence intervals Like extremely
a a 7 bc bc cd
Like moderately b bcd d bcd d Neither like nor dislike b bc
Taste
Dislike moderately
1 Melrose Spartan Lodel Rajka Topaz Rubinola Well known cultivars Scab resistant cultivars
Dislike extremely
Fig. 3. Comparison of taste of well-known and scab resistant apples means marked with the same letters do not differ significantly at P=0.05
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Fig. 4. PCA projection of profile sensory analysis for well-known ( ) and scab resistant ( ) cultivars subjected to consumer tests in 2004 and 2005. Attributes which differentiated the evaluated cultivars: aroma of ripe apples (2), firmness (6), juiciness (7), sweetness (8), sourness (9), astringency (10), flavour of ripe apple (11), others flavour (12) and over-all quality (13). Length of vectors joining attribute points with mid of co-ordinate system indicates significance of particular trait in quality differentiation.
Taking into consideration labour requirements and high costs of consumer tests it was interesting to check whether consumer appreciation of apple taste could be predicted by expert panel in laboratory tests. For this purpose the results of consumer taste assessment were compared with the overall quality evaluated by expert panel (Fig. 5). Both results seem to show good liner correlation (r = 0.72). This fact indicates that laboratory evaluation of the overall quality may be used as a prognostic indicator of consumer acceptance of apples among adult population.
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8
Consumer assessment of taste (points)
r = 0.722 **
7
MELROSE' 04
MELROSE' 05
RAJKA' 05 RUBINOLA' 04 RUBINOLA' 05 SPARTAN' 05 SPARTAN' 04 RAJKA' 04 TOPAZ' 04 LODEL' 04 LODEL' 05 TOPAZ' 05
Over-all quality (points)
Fig. 5. Relationship between overall quality evaluated by expert panel and taste assessed in consumer tests. Data for six apple varieties in two successive seasons are presented. Acknowledgement The research was performed within HIDRAS EU Project with the financial support from the Commission of the European Communities (contract N QLK5-CT-200201492). REFERENCES Baker G.A. 1999. Consumer preferences for food safety attributes in fresh apples: market segment, consumer characteristics, and marketing opportunities. J. Agric. Resource Economics 24: 80-97. Baker G.A., Crosbie P.J. 1994. Consumer preferences for food safety attributes: a market segment approach. Agribusiness: An Int. Journal 10(4): 319-324. Konopacka D., Pocharski W.J. 2001. The relationship between firmness and juiciness of Elstar, Gloster and Jonagold apples. J. Fruit Ornam. Plant Res. IX, 1-4: 9-17. Kuhn B.F., Thypo A.K. 2001. Sensory quality of scabresistant apple cultivars. Postharvest Biol. Techn. 23: 41-50. Luning P.A., Marcelis W.J., Jongen W.M.F. 2002. Food quality management. A techno managerial approach. Wageningen Press., Wageningen. Steenkamp J.B., Van Trijp H.C.M. 1996. Quality Guidance: A Consumer - Based Approach to Food Quality Improvement using Partial Least Squares. European Review of Agricultural Economics 23: 195-215.
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PORWNANIE JAKOCI JABEK ODMIAN TRADYCYJNYCH I PARCHOOPORNYCH W OCENIE EKSPERTW I KONSUMENTW Streszczenie Celem bada byo porwnanie jakoci jabek odmian dobrze znanych i tradycyjnie uprawianych (standardowych) oraz odpornych na parcha. Na podstawie obrazu PCA dla profilowej oceny sensorycznej wykonanej przez ekspertw dla 12 odmian standardowych i 5 parchooodpornych, do testw konsumenckich wybrano po 3 odmiany standardowe i odporne na parcha, charakteryzujce si wysok ocen jakoci, ale zrnicowane pod wzgldem cech sensorycznych. Wybrane odmiany to Lodel, Melrose i Spartan (odmiany standardowe) oraz Rajka, Rubinola i Topaz (parchoodporne). Testy konsumenckie przeprowadzano w sezonie 2004 i 2005 wrd dorosych mieszkacw Skierniewic. Wygld i smakowito jabek oceniano w skali hedonicznej. Wygld zewntrzny odmian parchoodpornych wydaje si by bardziej podatny na zmienno sezonow ni odmian standardowych. W roku 2004 odmiany parchoodporne byy postrzegane jako mniej atrakcyjne ni standardowe, podczas gdy w 2005 ich wygld by bardziej atrakcyjny i rnice pomidzy grupami stay si nieistotne. W przypadku oceny smakowitoci w obu sezonach odmiany parchoodporne byy zwykle oceniane niej ni standardowe, szczeglnie w porwnaniu do Melrose. Inne odmiany standardowe byy zwykle oceniane pomidzy 5 i 6 w skali 1-9 punktowej. Parchoodporna odmiana Rubinola bya postrzegana na tym samym poziomie jak standardowy Spartan. Najnisze oceny za smakowito uzyskiway Topaz (odmiana parchoodporna) i Lodel (standardowa). Ocena ekspertw wskazuje, e wysoka jdrno, soczysto, smakowito dojrzaych jabek zwikszaj akceptacj jakoci jabek w ocenie konsumentw. Ponadto stwierdzono, e ocena konsumentw jest istotnie skorelowana z ocen ekspertw (r = 0,72).

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EFFECT OF DIFFERENT FOLIAR SPRAYS ON THE STORAGE POTENTIAL AND FRUIT QUALITY OF CONFERENCE PEARS
Maciej GSTO1, Jan BASZCZYK1, Iwona DOMAGAA-WITKIEWICZ2 1 Department of Pomology and Apiculture 2 Department of Soil Cultivation and Fertilization Agricultural University Al. 29 Listopada 54, 31-425 Krakw, Poland e-mail: rogastol@cyf-kr.edu.pl
Summary The experiment was carried out in years 2004-2005. The effect of different foliar fertilizers (calcium chloride, Kalcisal, Kalcisal+Kalcifos and Sanisal) on fruit quality as well as on calcium content of Conference pear was investigated. The treatments consisted of five foliar sprays (at the rate of 0.5%) in two week intervals followed by five (at the rate of 1.0%) in one week intervals. At harvest and, additionally after 150 days of cold storage at 0C, fruits were analysed for firmness, soluble solids content (SSC) and titratable acidity (TA). The occurrence of fungal diseases was also investigated. Samples of fruits were mineralized and the content of calcium and potassium was assessed using atomic absorption spectroscopy. The highest fruit firmness, both at harvest and after the storage, was noted for these treated with Kalcisal+Kalcifos. Pears sprayed with calcium chloride revealed the highest SSC. The investigated foliar fertilizers significantly affected TA. At harvest, the lowest TA was noted for pears sprayed with Sanisal (0.19%), whereas the highest for CaCl2 (0.23%). The lowest acidity reduction during the storage for Kalcisal+Kalcifos was observed. All of investigated fertilizers had a beneficial effect on fungal diseases reduction. Calcium content was very differentiated and varied from 75.9 mgkg-1 f.w. (Control) to 101.3 mgkg-1 f.w. (CaCl2). key words: fruit quality, pears, calcium, fungal diseases INTRODUCTION Mineral content is one of major factors determining quality and storability of fruits (Bramlage et al. 1993). Calcium is of special importance, influencing many metabolic pathways. It increases cell wall resistance and rigidity, this way decreasing susceptibility to storage disorders (Marshner 1988). High Ca fruit content leads to decreasing respiration rates and delays senescence (Poovaiah et
al. 1993, Tomala & Trzak 1994). Increasing of fruit calcium content by spraying with calcium salt during fruit development leads to an increase in the fruit firmness (Cooper & Bangerth 1976). The most common way of improving Ca fruit level is use of foliar fertilizer, however the efficiency of the treatment depends on many factors, such as: timing, number of treatments, weather conditions and chemical formulation. Therefore, the objectives of the present work was: to determine the effect of different foliar fertilizers on flesh firmness, soluble solids content, acidity and fruit decay and to assess the effectiveness of these fertilizer in respect of increasing Ca fruit content. MATERIAL AND METHODS The experiment was conducted in years 2004-2005 in pear orchard at the Experimental Station in Garlica Murowana, near Krakw (Poland). The soil of the plot where the fruit trees were planted was in the valuation class II b. It is classified as a brown soil type developed from loess and represents a type described as silt loam. In the middle of July soil samples were taken for analysis, separately from the herbicide strips (HS), grass strips (GS), as well as from the layers of 0-20 and 20-40 cm in depth. Soil pH was determined in H2O and 1 M KCl. Potassium, magnesium and calcium were extracted according to the universal method (with 0.03 M CH3COOH) and measured by atomic absorption spectroscopy (Ostrowska et al. 1991) (Table 1).
Table 1. Contents of P, K, Mg and Ca (mgdm-3) in orchard soils pHH2O Herbicide/ Grass strips Soil layer depth (cm) HS GS 0-20 20-40 5.22 6.19 5.83 5.58 pHKCl 4.23 5.36 4.96 4.63 K 99.3 129.0 146.5 81.8 Mg 61.8 84.8 83.6 63.0 K/Mg 1.6 1.5 1.7 1.3 Ca 452.6 610.4 542.0 520.9
The plant material was composed of 16-year old Conference trees grafted on Pyrus caucasica rootstock. In the orchard, the soil cultivation system was a herbicidal fallow in rows and grass in the inter-rows. The pear trees were spaced 3.04.0 m. The crowns of trees were trimmed in a spindle form. The protection of the trees was carried out according to the recommendations accepted for commercial orchards. The experiment was established in a randomized block design, each treatment being represented by five replications plots of two trees each. The following treatments were used in the experiment: 1. control trees sprayed with water, 2. calcium chloride, 3. Kalcisal (11% Ca, 0.1% Mg, 0.02% B), 4. Kalcisal + Kalcifos (2% Ca, 18% P, Mg 0.1%, 0.02% B), 5. Sanisal (kaolin clay).
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The treatments consisted of five foliar sprays (at the rate of 0.5%) at two week intervals followed by five (at the rate of 1.0%) at one week interval. At harvest 20 pears for each plot were assessed for: firmness (8 mm plunger), soluble solids content and titratable acidity. The same analyses were performed after the cold storage for 150 days at 0C and relativ air humidity of 90-92%. Additionally, the incidence of fungal diseases was investigated. Fruits from the treatments were washed and ten pears were taken for mineral content analyses. Stems and seeds were removed. Fruit flesh was mineralized in a mixture of HNO3:HClO4:H2SO4 (6:2:0.25) and analyzed using atomic absorption spectroscopy (AAS) to assess the content of potassium and calcium. The measurements were listed and subjected to analysis of variance. Differences between the means were ascertained with a multiple Duncan Test, using a Statistica 6.0 program. The mean values for the combinations labeled with the same letters do not significantly differ at the significance level P=0.05. RESULTS AND DISCUSSION The effect of foliar fertilizers on investigated quality indicators, such as: firmness, soluble solids content and titratable acidity was significantly affected by the weather conditions during the vegetative period. As far as fruit firmness is concerned, a beneficial effect of calcium fertilizers was observed, both at harvest and after the storage. This confirms the previous report (Raese 1994) that calcium treatment increased fruit firmness. However, the effect on firmness was really small (although statistically significant). The same revealed Frances et al. (1999) - intensive calcium treatments had only a slight effect on firmness retention. Sanisal treatment did not influenced pears firmness at harvest, though it reduced fruit softening during the storage (Table 2). Mean soluble solids concentration, measured for control fruits at harvest time, was higher as compared with the rest of treatments. The only exception was CaCl2. Pears treated with CaCl2 maintained the highest SSC also after storage (Table 3). Fertilizers had no or little effect on TA when measured at harvest (Table 4). Only fruits treated with Sanisal revealed lower TA. The lowest reduction of titratable acidity after storage was found for fruits sprayed with Kalcisal+Klacifos. The lowest TA level after the storage time was recorded for control fruits and calcium chloride. During the experiment the incidence of blue mould (Penicillium expansum) and grey mould (Botritis cinerea) was observed. However, the percent of infected fruits was low (Fig. 1). Calcium fertilizers significantly reduced incidence of these fungal diseases. The only exception was CaCl2. Surprisingly, this treatment increased percent of infected fruits, but only in the second year of the experiment. This was mainly connected with stronger blue mould infection. Mean potassium and calcium content was strongly affected by the weather conditions. In 2004 pears contained higher K and Ca concentration, whereas the higher K/Ca ratio was noted in 2005 (Table 5). The mineral content was also
significantly affected by foliar fertilizers. In 2004, the highest Ca level was found for trees sprayed with Kalcisal (125.5 mg Ca kg-1 f.w.), whereas in 2005 with calcium chloride (101.3 mg Ca kg-1 f.w). K/Ca ratio varied from 16-35 and 21-35, for year 2004 and 2005, respectively. As far as K/Ca proportion is concerned, fruits treated CaCl2 and Kalcisal did not differ as compared with control ones. Fruits from trees sprayed with Kalcisal+Kalcifos and Sanisal revealed high K/Ca ratio.
Table 2. Firmness (kG) of Conference pears as affected by foliar fertilizers Treatment Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal At harvest 2005 Mean for 2004-2005 6.77 a 6.90 a 6.94 a 7.16 bc 6.85 a 7.13 b 7.55 b 7.37 c 6.73 a 6.86 a After storage 2.87 a 4.10 a 3.57 b 4.44 bc 3.03a 4.47 bcd 4.13 c 4.62 e 3.99 c 4.86 cd
2004 7.04 a 7.37 b 7.41 b 7.18 ab 6.98 a 5.33 ab 5.30 ab 5.91 c 5.58 b 5.25 a
Note: Mean separation according to Duncan multiple range test. Numbers followed by the same letter within the same column and the analysis time are not different at P=0.05
Table 3. Soluble solids content (Brix) of Conference pears as affected by foliar fertilizers Treatment Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal Note: see Table 2. At harvest 2005 Mean for 2004-2005 13.50 d 12.85 d 13.90 e 13.08 e 13.20 c 12.48 c 12.50 a 12.33 b 12.60 b 12.23 a After storage 12.70 a 13.30 a 15.07 e 14.73 c 14.20 d 13.70 ab 13.97 c 14.10 b 13.57 b 13.38 a
2004 12.20 b 12.27 b 11.77 a 12.17 b 11.87 a 13.90 b 14.40 c 13.20 a 14.23 c 13.20 a
Table 4. Titratable acidity (% of malic acid equivalents) of Conference pears as affected by foliar fertilizers Treatment Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal Note: see Table 2. At harvest 2005 Mean for 2004-2005 0.191 a 0.211 bc 0.200 bc 0.231 c 0.211 b 0.217 bc 0.198 b 0.202 b 0.202 c 0.194 a After storage 0.158 b 0.142 a 0.167 c 0.140 a 0.134 a 0.147 b 0.173 c 0.165 d 0.185 d 0.160 c
2004 0.232 d 0.262 e 0.223 c 0.207 b 0.187 a 0.126 b 0.113 a 0.160 e 0.156 d 0.136 c
7 Year 2004 6 Year 2005
LSD(P=0.05) Year = 0.320 Factor: Treatment = 0.507 Year x Treatment = 0.717
% of diseases
0 Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal
Treatment
Fig. 1. Fungal diseases incidence of Conference pears as affected by different foliar sprays
Table 5. Potassium and calcium fruit content (mg kg-1 f.w.) and K/Ca proportion as affected by different foliar fertilizers in years 2004-2005 Treatment Control CaCl2 Kalcisal Kalcisal+Kalcifos Sanisal Mean for year LSDYear (P=0.05) Note: see Table 2. K 2004 2005 1380 a 1228 a 1374 a 1317 ab 1655 c 1258 ab 1757 b 1544 c 1800 b 1363 b 1593 1342 45.5 Ca 2004 2005 108.8 b 75.9 b 113.8 b 101.3 c 125.5 c 92.8 a 89.1 a 95.0 a 91.6 a 91.1 a 105.8 91.2 2.50 K/Ca 2004 17 a 16 a 18 a 30 b 35 c 23.4 1.66 2005 25 ab 21 a 21 a 35 c 28 b 26.0
CONCLUSIONS Quality parameters of Conference fruits as well as their mineral content were strongly affected by weather conditions. Calcium fertilizers had a beneficial effect on firmness (both at harvest and after the storage) and overall fungal diseases incidence. With an exception of CaCl2 used fertilizers significantly slowed down titratable acidity reduction. Treatments also affected the soluble solids content fruits treated with calcium salts revealed higher SSC after the storage. Calcium chloride and Kalcisal treatment significantly increased Ca fruit content. Although Sanisal and Kalcisal+Kalcifos favoured high K/Ca ratio (2835), no physiological disorders for these treatments were observed.
REFERENCES Bramlage W.J., Barden C.L., Watkins C.B. 1993. Comparing potential predictors of scald susceptibility of apples (Malus domestica Borkh.). Acta Hort. 326: 237-244. Cooper H.D., Bangerth F. 1976 The effect of Ca and Mg treatment on the physiology, chemical composition and bitter pit development. Sci. Hort. 5: 49-57. Frances J., Juan J.L., Montesinos E., Vilardell P. 1999. Minimised post-harvest chemical treatments, fruit density per tree and calcium sprays affect the storability of Passe Crassane and Conference pears in Girona (Spain). Acta Hort. 485: 161-166. Marschner H. 1988. Mineral nutrition of higher plants. Acadmic Press Ltd., London. Ostrowska A., Gawliski S., Szczubiaaka Z. 1991. Metody analizy i oceny waciwoci gleb i rolin. Instytut Ochrony rodowiska, Warszawa. [in Polish] Poovaiah J.W. 1993. Biochemical and molecular aspects of calcium action. Acta Hort. 326: 139-147. Raese J.T. 1994. Effect of calcium sprays on control of black end, fruit quality, yield and mineral composition of Barlett pears. Acta Hort. 367: 314-322. Tomala K., Trzak M. 1994. Occurence of cork spot (pit) in Alexander Lucas pears depends on fruit mineral element content. Acta Hort. 368: 570-577.
WPYW NAWOENIA DOLISTNEGO NA JAKO I WACIWOCI PRZECHOWALNICZE GRUSZEK ODMIANY KONFERENCJA Streszczenie Celem dowiadczenia prowadzonego w latach 2004-2005 bya ocena wpywu nawozw dolistnych (chlorek wapnia, Kalcisal, Kalcisal+Kalcifos i Sanisal) na parametry jakociowe i zawarto wapnia w owocach. Drzewa opryskiwane byy piciokrotnie roztworami nawozw w steniu 0,5% w odstpach dwutygodniowych, a nastpnie piciokrotnie roztworami 1,0% w odstpach tygodniowych. Bezporednio po zbiorze oraz po 150 dniach przechowywania gruszek w chodni zwykej (temp. 0C) zmierzono ich jdrno, zawarto ekstraktu i kwasowo. Okrelono take stopie poraenia owocw chorobami grzybowymi. W zmineralizowanych prbkach owocw okrelona zostaa zawarto wapnia. Najwysz jdrno, zarwno po zbiorze, jak i po przechowywaniu odnotowano dla owocw traktowanych Kalcisalem+Kalcifosem. Gruszki opryskiwane chlorkiem wapnia posiaday najwysz zawarto ekstraktu. Zastosowane nawozy dolistne istotnie wpyny na kwasowo owocw. Po zbiorze najnisz kwasowo zmierzono w gruszkach opryskiwanych Sanisalem (0,19%), podczas gdy najwysz chlorkiem wapnia (0,23 %). Najniszy spadek zawartoci kwasw w trakcie przechowywania zaobserwowano dla kombinacji Kalcisal+Klacifos. Wszystkie z zastosowanych nawozw korzystnie wpyny na obnienie udziau owocw poraonych przez choroby grzybowe. Zawarto wapnia w owocach bya silnie zrnicowana i wahaa si od 75,9 mgkg-1 w.m. w kontroli do 101,3 mgkg-1 w.m. w traktowanych chlorkiem wapnia.

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PL ISSN 1506-9427

Vegetable Crops Research Bulletin

RESEARCH INSTITUTE OF VEGETABLE CROPS INSTYTUT WARZYWNICTWA SKIERNIEWICE, POLAND

Copyright by RIVC Printed in RIVC Skierniewice, Poland

Instructions for authors of publications in Vegetable Crops Research Bulletin

2006 VEGETABLE CROPS RESEARCH BULLETIN 65

FUSARIUM BASAL ROT IN THE NETHERLANDS

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Fig. 1. A field with Fusarium basal rot

Fig. 2. Onion bulbs with Fusarium basal rot symptoms

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infection level of the soil

Fig. 3. Influence of plant condition on infection

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Fusarium at harvest (%)

Bio-assay result (%)

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Soaking in excessive water amount

Soaking in limited water amount

Control 30% m.c. 35% m.c. 40% m.c.

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-0.427 xx -0.027 0.018

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VEGETABLES SEEDS NUTRITIONAL ASPECTS IN RESPONSE TO GERMINATION TIME

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Germination time (days)

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0% Raw 1 3 5 Germination time (days)

Fig. 2. Chemical composition of raw and germinated alfalfa seeds

Germination time (days)

Fig. 3. Chemical composition of raw and germinated radish seeds

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