INFLUENCE OF DURATION OF STORAGE ON VIABILITY, GERMINATION AND SEEDLING GROWTH OF Treculia africana Decne

BY

Nuga, O.O*, Okonkwo, H. O**, and Deckor, A*,

*Department of Forestry and Wildlife Management, Faculty of Agriculture, University of Port-Harcourt, P.M.B.5323, Port-Harcourt, Nigeria

** Swamp Forest Research Station, Onne P.M.B 006 Eleme-Nchia, Rivers State, Nigeria

Corresponding Author: Okonkwo, H.O E- mail: richychrist4ever@yahoo.com

ABSTRACT
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Treculia africana is an important, multi-purpose, under-utilised, indigenous tree species in Nigeria and in Africa as a whole. The seeds are widely consumed in these parts especially by rural dwellers thus, making the species a food security and rural livelihoods enhancer. The seeds of the species are however recalcitrant in nature and poses storage problems. The influence of duration of storage on seed viability, germination and seedling growth of Treculia africana were investigated. Seeds were stored for 7, 14, 21, and 28 days and were subsequently subjected to viability tests and germination trials at the end of each storage period. Seedlings from each of the seed treatments were further assessed to determine the effect of duration of storage on seedling growth. Some seeds were however sown immediately after extraction and these served as control. Viability tests results showed that control seeds had the highest percentage viability of 98% while seeds stored for 28 days had the least with 94%. Seeds stored for 7, 14, and 21days had 97%, 96%, and 95% viability respectively. Germination commenced 19days after sowing (DAS) in control seeds, while seeds stored for 7, 14, 21, and 28days germinated on the 22 nd , 28th ,29th ,and 32nd day respectively. Also, control seeds had the highest cumulative percentage germination (CPG) of 46% followed by 7days stored seeds with 28% germination. Seeds stored for 21days had 23%, while the ones stored for 28days had the least germination with 21%. The seeds stored for 14days were however exhumed by rodents from the germination tray. A progressive decrease in germination of the species seeds was observed as storage duration increased. In addition, storage duration had significant effect (p<0.05) on height and diameter growth of seedlings but showed no significant difference on the number of leaves. Seedlings of the control seeds had the highest number of leaves while seedlings from 7days stored seeds had the tallest plants and seedlings from 21, and 28days stored seeds had the best diameter growth. The study suggests that Treculia africana seeds should be sown immediately after extraction from the pulp. Where storage is necessary, however, it should be for just a week, if the storage method is such that exposes the seed to changes in temperature and humidity like the one used in the study. Sowing immediately after extraction will also enhance good foliage production where the growers intent is provision of fodder.

INTRODUCTION
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Seeds of several tropical fruit crops show recalcitrant storage behaviour. The seeds do not withstand drying or are unable to survive low temperatures during storage. Thus they are difficult to store for longer periods as noted by Ellis and Hanson (1984). The exact causes of recalcitrant seed death and its relationship with moisture content are not fully understood (Fu, et al,1993) even though loss of viability could either be caused by the moisture content falling below a certain critical value or simply a general physiological deterioration with time as pointed out by (Chin, et al, 1984). However , Hanson(1984) noted that several pre-harvest factors including cumulative effect of environment during seed maturation, harvesting, drying and the pre-storage environment, time of seed harvesting, duration of drying and the subsequent period before seed is placed in store determine the longevity of seeds. Onyekwelu and Fayose (2007) positioned that large volumes of high quality seeds are lost for planting yearly because of inadequate drying and storage of seeds. Treculia africana (African bread fruit) is a large evergreen tropical food tree species belonging to the Moraceae family (Onyekwelu and Fayose, 2007). It is popularly called Ukwa by the Igbo tribal group of south-eastern Nigeria. The plant produce large compound fruits (usually round) which are covered with rough pointed outgrowth and has the seeds buried in spongy pulp of the fruits (Keay, 1989). Three varieties (africana, inversa, and mollis) have been distinguished (Okafor, 1990). Their taxonomic differences are mainly based on fruit-head size, hairiness of branchlets and leaves. Variant africana produces fewer and larger fruit with superior seed weight while variant inversa produce twice as many branches than variant africana (Okafor 1990; WAC, 2005). The seeds of African bread fruit are highly nutritious and constitute a cheap source of vitamins, minerals, proteins, carbohydrates and fat (Okafor and Okolo, 1974). The seeds have an excellent polyvalent dietetic value, the biological value of its protein exceeding even that of soybeans (WAC, 2005). The seeds are widely consumed, thus playing important role in food security, especially among rural dwellers in West Africa (Onyekwelu and Fayose, 2007). Many rural dwellers in Nigeria and Cameroon are said to be engaged in collection, processing and sale of seeds of the species as a means of livelihood. Despite the socio-economic importance of Treculia africana to a very large population of people and particularly, people in south-eastern Nigeria, a lot of seeds are being lost annually to improper storage (in terms of duration of storage) and this is greatly hampering the regular cultivation and conservation of the species thereby pre-disposing it to extinction.
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Purohit and Doijode (1998) noted that germplasm can be conserved in several ways but conservation in the form of seeds has many advantages as it is simple to use, easy to handle, practicable, inexpensive and capable of maintaining genetic stability during storage. If seeds are not properly stored in terms of the required length of time they stand to lose viability as the length of time seeds will remain viable varies greatly by species and storage conditions (Onyekwelu and Fayose, 2007). There is information gap on the length of time for which Treculia africana seeds stay viable in storage on the basis of being a recalcitrant species and for the purpose of germplasm conservation, hence, the aims of this investigation were (i) to confirm the existence of recalcitrant storage behaviour of the seeds and (ii) to determine the effect of storage time on seed viability and germination as well as seedling growth.

MATERIALS AND METHODS

Fruit Collection and Processing The fruits of Treculia africana used for this study were collected from the Swamp Forest Research Station, Onne, Rivers State. The area is located on Lat. 4 0 410 23 N and 70 091 28 E. Annual rainfall is 2500mm with a mean value of 75% relative humidity in February and 80% in July. The mean minimum temperature is 250C (ICRAF/IITA, 1994). While, the germination trials were conducted at the nursery site of the Department of Forestry and Wildlife Management, Faculty of Agriculture, University of Port-Harcourt, Choba, Rivers State. The area is located on Lat. 40 53I N and Long. 60 54I N. The fruits were allowed to deteriorate for two (2) weeks to ease seed extraction. They were de-pulped on the expiration of the period by washing the already fermented pulp in tap water and the seed removed. Viability Test To ensure that the seeds used in the experiment were high quality and of the same viability status, the sample lot was subjected to viability test using the floatation test (Onyekwelu and Fayose, 2007). The seeds were soaked in a bucket half filled with water for twenty-four (24) hours. The seeds that floated on the water on the expiration of the soaking period were taken as unviable and discarded while those that sank to the bottom of the bucket were collected and used for the experiment. Storage periods
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Sample seeds used in the control experiment were sown immediately after seed extraction. Subsequently the seeds were sun-dried for three hours, airdried for three (3) days at room temperature (270C) and finally stored in a perforated metal container. Four (4) storage treatments 7, 14, 21, and 28days were used in the experiment. One hundred sample seeds were collected from the lot at the expiration of each storage regime and tested for viability, germination and growth performance. Germination Studies The germination medium was sharp river sand. Watering was done once each day. Plumule emergence was taken as an indication of germination. Germination count lasted sixty (60) days. Factors determined were the germination date, rate and duration of germination. Growth studies Thirty (30) uniformly growing seeds were randomly selected from the seedlings of germinated seeds. These were transplanted after a period of sixty (60) days into poly pots filled with top soil and of same sizes. After a week of weaning in the green house the seedlings were laid out under a partially shaded shed for data collection. The experimental layout was a Completely Randomised Design replicated four times. Collar diameter, height, and number of leaves were the growth factors evaluated. Height was measured with a simple transparent 30cm ruler, collar diameter with an electronic digital veneer calliper with a measuring range; 0-150mm, 0-200mm, 0-300mm, and leaf were counted. Statistical analysis Germination rate was calculated using a simple calculator, analysis of variance (ANOVA) was used to test for significant differences between and among the growth factors evaluated and Duncan Multiple Range test was used for mean separation.

RESULTS AND DISCUSSION

Seeds in the control experiment showed the highest percentage viability. Subsequently there was a gradual but continuous decline in the viability of the stored seeds as the storage duration increased (Fig. I).
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Fig. I: Influence of storage duration on the viability of Treculia africana seed Germination commenced earlier in the control seeds than in the stored seeds (Table I). Similarly percentage germination was highest in the control when compared to those of the seeds subjected to storage treatment. After the first seven days of storage, almost half the germination capacity of the seed was lost, dropping from 46 to 28%. By the 28 th storage day, which is barely a month, the percentage germination had reduced to 21%. This is equivalent to losing more than half of a batch of seed within less than a month of storage. The fourteen (14) days stored seeds were exhumed from the germination tray by rodents in the course of the experiment, as a result, data was not available for the calculation of the percentage germination and subsequent growth analysis was hampered. Onyekwelu and Fayose, (2007) reported that apart from reduction in viability and germination capacity, storage also reduces the nutritive value of Treculia africana seeds. Moreover, Fig. II, show a reduction in vitality or germination rate, as well, with storage duration. According to Nwoboshi (1982) seeds with low rate of germination tend to produce weak and sometimes abnormal seedlings.

Fig. II: Influence of storage duration on the germination of Treculia africana seed

The main aim of seed storage is to maintain the viability of the seed at the highest level possible after storage period. Some species, however, are short-lived and normally lose their viability rapidly even within a few days or months of collection irrespective of the storage conditions (Nwoboshi, 1982). Treculia africana seed has been classified among the inherently short-lived tropical species (Onyekwelu and Fayose, 2007). The study therefore confirms the observation. The particular factor responsible for the behaviour, Nwoboshi, (1982) reported is the seed coat, long-lived seeds have been observed to have hard seed coats, while the short-lived or recalcitrant ones do not. Seeds with hard impermeable seed coats, if highly viable and undamaged its life span will hinge primarily on species characteristics and the temperature and humidity condition prevalent during storage.

Table I: Influence of storage on the Cumulative Germination Percentage of Treculia africana seed

Storage Duration (days)

Germination day

CGP (%)

Germination days

0 (Control) 7 14 21 28

19 22 28 29 32

46 28 23 21

41 39 32 29

Moreover, several authors (Harrington, 1972; Nwoboshi, 1982; Onyekwelu and Fayose, 2007) have observed that seed moisture content plays important role in maintaining seed viability during storage. Low seed moisture is favourable for keeping seed viability since it leads to a reduction in the rate of vital physiological processes and controls the insect and microflora infestation of the seed. Therefore, there is a need to dry seeds before storage. The danger here, however, is that of over-drying which can be harmful to the seed if the critical moisture content level is exceeded for the species in question. Since the critical seed moisture level varies with species, the solution to seed viability loss during storage of Treculia africana may lie in working out the value for the species together with the relative humidity of the storage atmosphere. The result of the growth data show that seedlings from the seven (7) days stored seed performed better in height compared to those from the control seeds, followed by 21days, and the 28days respectively (Fig. III). This is contrary to the observations in the viability and the germination tests results in which the control performed better than the stored seeds. Since the seven (7) days stored seeds germinated three (3) days after the control (Table I), the expectation was that the control should perform better in growth than the seedlings from the stored seeds. However the results show a contrary development. Analysis of variance reveal a significant difference (p>0.05) in height growth among the storage treatments. However, Duncan multiple range test reveal that there is no significant difference (p>0.05) in height between seedlings of 21days and 28days stored seeds. This is suggestive of a relationship between short term seed storage and early height growth of seedlings of the species. Seedlings of the two longest stored seeds 21 and 28days showed better diameter growth compared to those from the control and the 7days stored seeds (Fig. III). Seedlings from
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7days stored seeds recorded higher diameter growth than the control. Duncan multiple range test, however, suggests there is no significant difference (p>0.05) in diameter growth between the seedlings of the control and the 7days. The result suggests a relationship between diameter growth and storage duration between 21 and 28days. Control seedlings recorded highest number of leaves, followed immediately by the 7days, the 28days and the 21days respectively (Fig. III). Hence little or no storage translates to leaf accumulation in early growth of the species. Analysis of variance, however, shows that there was no significant difference (p>0.05) in leaf number among the four (4) storage treatments.

Fig. III: Influence of storage duration on the early growth of Treculia africana seedlings

According to Nwoboshi, (1982) and Ekeke (1995) seedling vigour or the ability of the young plants to grow rapidly and withstand environmental stresses tends to be greatest in seeds with high rates of germination, while embryos with a low rate of germination tend to produce weak and sometimes abnormal seedlings that are less able to withstand unfavourable environmental conditions. Apart from its performance in leave number, the growth of the control seedlings in height and diameter completely contradicts the observation of the two authors cited. The same control seeds that demonstrated highest viability and germination percentage produced seedlings of lesser vigour compared with those from the stored seeds which had earlier performed poorly in viability and germination capacity. Although the
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finding is subject to further scrutiny as other factors could contribute to the development, an example is variation in soil fertility. The conclusion of the study is that storage duration has effect on the early growth of Treculia africana seedlings.

CONCLUSION
The findings of the study confirms the earlier observations of FAO (1958) and Nwoboshi (1982) that seeds which do not possess hard impervious seed coats like Treculia africana, lose their viability rapidly as they are subject to temperature and humidity fluctuations. Closed containers are likely to be more suitable for the storage of the species. Further studies, therefore is going to be directed at determining which container type best stores Treculia africana seeds and for how long can it be stored with only a minimum loss of viability and germination energy. The study suggests that Treculia africana seeds should be sown immediately after extraction from the pulp. Where storage is necessary, however, it should be for just a week, if the storage method is such that exposes the seed to changes in temperature and humidity like the one used in the study. Sowing immediately after extraction will also enhance good foliage production where the growers intent is provision of fodder.

REFERENCES
Chin, H.F., Hor, Y. L., and Mohammed Lassim, M.B (1984): Identification of recalcitrant seeds. Seed Science and Technology: 12:429-436. Ekeke, (1995): Germination of Chrysophyllum albidium and Denneltia tripetala. Nig. J. Agric. Teacher Educ. IV (2): 179-185. VoTeX Publishers. Ellis, R. H (1984): Revised table of seed storage characteristics. Plant genetic new 1: 58: 1633.
FAO (1958): Choice of species. F.A.O. Forestry Development Paper No.13

Fu, J. R., Xia, O.H., Tang, L.F. (1993): Effects of domestication on excised embryonic axes of three recalcitrant seeds and studies on cryo-preservation seed science and technology: 21:85-95. Hanson, J. (1984): The storage of Tropical Fruit Trees. In: Crop genetic resources conservation and evaluation (J.H.W. Holden and William, eds) Goerge Allen and Unwin, London: Pp. 53-62.

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Harrington, J.F., (1972): Seed storage and longevity. In (Kozlowski T.T. ed) Seed biology Vol. 13: Insects, and seed collection, storage, testing and certification, Academic press N.Y.: 145-245. ICRAF/IITA (1994): Annual Report. International Centre for Research In Agroforestry/Tree improvement project, Onne, Nigeria: Pp. 50. Keay, R.W.J., (1989): Trees of Nigeria: A revised version of Nigerian Trees Vols. 1&2 Stanfield Dp, Clarendon Press, Oxford. Nwoboshi, L.C., (1982): Tropical Silviculture (Principles and techniques). Ibadan University Press Publishing House, University of Ibadan, Ibadan, Nigeria Okafor, J.C., (1990): Indigenous Trees of the Nigerian Rainforest. A paper presented in a Symposium on the Potentials of Domesticating and rebuilding Forest Resources. Yaounde Cameroon: Pp. 34-38. Okafor, J.C., and Okolo, H.C., (1974): Potentials of some indigenous fruit trees of Nigeria. Paper presented at the fifth Annual Conference of the forestry Association of Nigeria, Jos. Onyekwelu, J.C., and Fayose, O.J., (2007): Effect of storage methods on the germination and proximate composition of Treculia africana seed. Paper presented at the conference on International Agricultural Research for Development. Tropentas, Germany. Purohit, A.G., and Doijode., S.D., (1998): Activities at IIHR: Germplasm introduction, exchange and conservation with emphasis on fruit crops. In: tropical fruits in Asia Diversity, Maintenance, conservation and use (Arora, R.K., and Ramanatha Rac., V. Eds). Proceedings of the Ipgri-kar Utfanet Regional Training Course on the conservation and use of germplasm of tropical fruits in Asia held at Uhr, Bangalore, India: May 18-31, 1997. Pp 157 World Agroforestry Centre (WAC), (2005): Treculia africana in: Agroforestry Data Base. http:/www.WorldAgroforestry Spid=1651. org/Seg/Products/Addbases/At/Ass/Species/InforAsp?

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