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Journal of Plant Nutrition

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Seasonal Variation in Nutrient Status of Foxglove Leaves

Luis Roca-Prez , Rafael Boluda & Pedro PrezBermdez
a a a a

Departamento de Biologa Vegetal, Facultat de Farmcia, Universitat de Valncia, Valencia, Spain Available online: 22 Sep 2006

To cite this article: Luis Roca-Prez, Rafael Boluda & Pedro Prez-Bermdez (2006): Seasonal Variation in Nutrient Status of Foxglove Leaves, Journal of Plant Nutrition, 29:6, 1077-1084 To link to this article: http://dx.doi.org/10.1080/01904160600689191

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Journal of Plant Nutrition, 29: 10771084, 2006 Copyright Taylor & Francis Group, LLC ISSN: 0190-4167 print / 1532-4087 online DOI: 10.1080/01904160600689191

Seasonal Variation in Nutrient Status of Foxglove Leaves

Luis Roca-P rez, Rafael Boluda, and Pedro P rez-Bermudez e e
Departamento de Biologa Vegetal, Facultat de Farm` cia, Universitat de Val` ncia, a e Valencia, Spain

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ABSTRACT
The seasonal variation of mineral elements and the relationships among them were studied in natural populations of foxglove (Digitalis obscura). Young and mature leaves were collected in 10 different populations and on four sample dates (May, July, October, and February). Leaf mineral elements [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu)] were determined. The highest concentrations of N, P, and K in young leaf were recorded in May, followed by a decrease in the other months, while in contrast Ca and Fe showed the lowest concentration in May. Mature leaves showed differential seasonal behavior. Besides seasonal variations, signicant uctuations of N/P and Ca/Mg ratios were observed in young leaves. Strong positive correlations existed among N, P and K, while negative correlations were found between Ca and N, P, or K. Keywords: Digitalis, nutrients, seasonal, correlations, ratios

INTRODUCTION The analysis of the nutrient leaf status allows testing hypotheses involving plant nutrient concentration and different factors (climatic, soil nutrient availability, seasonal, leaf age, etc.), associated with the inter- or intraspecic variations observed. Besides individual element analyses, the ratios between nutrients are important for studying plant communities and their responses to environmental changes. In the case of seasonal variations in the plant nutrient concentrations and their ratios, several studies have shown that dilution and retranslocation are
Received 29 March 2005; accepted 4 January 2006. Address correspondence to Luis Roca-P rez, Departamento de Biologia Vegetal, e Facultat de Farm` cia, Universitat de Val` ncia, Av. Vicent Andr s Estell s s/n, 46100 a e e e Burjassot, Valencia, Spain. E-mail: luis.roca@uv.es 1077

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the main factors related to these uctuations (Thiec et al., 1995; Drossopoulos et al., 1996; Vemmos, 1999; G sewell, 2004). u On the other hand, there are large specic and nonspecic interactions among plant nutrients that, in some cases, could be characterized by correlation analysis between different nutrients (Garten, 1976; Meerts, 1997; Thompson et al., 1997; Trabaud, 2001; Maier and Chvyl, 2003). This work deals with Digitalis obscura L. (Scrophulariaceae), or foxglove, a perennial shrub found in the Iberian Peninsula whose importance lies in its capacity for cardiac glycosides biosynthesis. Previous works have revealed the possible relationships between mineral nutrition and cardenolide accumulation in natural plants of this species (Roca-P rez et al., 2002, 2004a, 2004b, e 2005); therefore, it is necessary to know the variations in the concentration of mineral elements, as they could affect the production of these secondary metabolites. On the basis of what has been stated, the objectives of this work were to study seasonal variations in mineral nutrients, as well as to determine the relationships between different elements in young and mature leaves of D. obscura plants growing in wild populations.

MATERIALS AND METHODS Sampling and Analyses Sampling took place throughout the year (in February, May, July, and October) from 10 natural populations of Digitalis obscura L. located in the Valencian Community (Spain): Ayora, Camporrobles, Llanorel, Garb, Olocau, Sinarcas, Toro, Tu jar, Vall dAlcal` -1, and Vall dAlcal` -2. The different stand charace a a teristics, and the physical and chemical properties of the soils under the studied plant populations are provided in Roca-P rez et al. (2004a). e Ten plants were selected from each population and for each sample date. Each sample consisted of 40 young leaves (the rst four underneath the apical bud), or 40 mature leaves (the rst four above the clearly senescent leaves) collected separately. The leaves were washed, dried, and pulverized. Samples were digested with nitric acid for the nutrient analysis. Potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) concentrations were measured by atomic absorption spectrometry (Perkin Elmer 2080); phosphorus (P) was determined by spectrophotometry; and the nitrogen (N) content was assessed by the standard Kjeldahl procedure. Accuracy of methods for acid digestions was veried by means of recovery assay using certied reference material from the Community Bureau of Reference, CRM 129 Hay Power, and CRM 281 Rye Grass. The recovery values for the analyzed elements were greater than 85%.

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Statistical Analysis Data were subjected to analyses of variance and means were compared by Tukeys test. Either the Pearson or Spearman correlation tests were used to establish relationships between mineral nutrients. All analyses were performed with the SPSS 9.0 program. RESULTS AND DISCUSSION Seasonal Variations in Nutrient Contents

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The results provided in Table 1 show the seasonal variations of macro- and micronutrient contents determined in D. obscura leaves. The quantity of macronutrients in the young leaves varied markedly among seasons (Table 1). Here, we observed that the leaves collected in May showed signicantly higher values of N, P, and K than those found during the other months; in contrast, Ca concentration was the lowest of the year in this particular month. Magnesium was the only macronutrient that did not present signicant seasonal variations in young leaf. In general, these data are in accordance with the results reported for other plant species, which indicated that the concentration of elements such as N (Vemmos, 1999), P (Thiec et al., 1995), and K (Drossopoulos et al., 1996) decreased throughout a seasonal cycle, whereas Ca content increased as a result of the leaf aging process (Drossopoulos et al., 1996; Vemmos, 1999).
Table 1 Seasonal variation of nutrients in young and mature D. obscura leaves Leaf type Young May July October February Mature May July October February % (DW) N P K Ca Mg Fe mg Kg1 (DW) Mn Cu Zn

2.48a 1.10b 1.22b 1.40b 1.03a 1.01a 1.12a 1.08a

0.29a 0.11bc 0.09c 0.14b 0.12a 0.09bc 0.08c 0.10ab

1.63a 0.92b 1.01b 1.04b 0.90a 0.93a 1.09a 0.91a

0.59c 0.79b 1.02a 0.99a 1.52a 1.12b 1.09b 1.15b

0.21a 0.20a 0.23a 0.21a 0.29a 0.25a 0.24a 0.23a

50b 105a 92a 82a 84b 172a 111b 110b

24a 25a 34a 31a 47a 35a 36a 36a

9bc 8c 11b 14a 9b 9b 11ab 13a

37a 26b 32ab 38a 52a 38a 40a 46a

For each leaf type and mineral element, values with the same letter represent a nonsignicant difference according to Tukeys test (p < 0.05). Data are the means of the 10 populations studied.

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With regard to the micronutrients in young leaves, Fe showed signicantly lower values in spring than those observed during the rest of year; Cu and Zn concentrations showed signicant differences among sampling dates, although they did not present a clear seasonal trend; and Mn foliar concentration did not undergo substantial changes (Table 1). These results are comparable to those for olive tree leaves by Fern ndez-Escobar et al. (1999), who reported a pattern a for Fe identical to that in D. obscura and varying seasonal proles for the rest of the microelements. In the case of mature leaf (Table 1), some mineral nutrients showed signicant differences among sampling dates, but seasonal trends were not found, which could be related to nutrient imbalances due to retranslocation and to an increased leaching of nutrients due to the fact that these leaves are presenescent organs. The results obtained suggest that the decrease in the N, P, and K percentages between May and July is due to an increase in the contents of structural materials (cellular wall) and other compounds such as starch, caused by the foliar-development process that takes place throughout spring and early summer. Moreover, the retranslocation of these mobile minerals to different sink organs (i.e., owers and fruits) could also contribute to this decrease, especially because the soils where D. obscura populations grow present low contents of N and P (Roca-P rez et al., 2005). On one hand, it is known that species from ine fertile environments tend to be more procient at retranslocating nutrients than those from fertile environments (Aerts and Chapin, 2000). On the other hand, despite the dilution effect, Ca and Fe contents in D. obscura leaves increased throughout the seasons due to the low or intermediate mobility in phloem of these elements and the high availability of these minerals in the soils studied (Roca-P rez et al., 2004a, 2005). e Comparisons of the nutrient contents in young and mature leaves showed a very low interpopulation variability in macronutrients (Roca-P rez et al., 2005) e and in micronutrients (Roca-P rez et al., 2004a). The only exception was Mg; in e plants from Ayora, content of this element was signicantly (P < 0.05, Tukeys test) higher than in the rest of the populations. In this plant population, the leaf contents were 0.36% and 0.44% dry weight in young and mature leaves, respectively.

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Variation in Nutrient Ratios Of the possible ratios between the mineral nutrients, only the seasonal variations of N/P and Ca/Mg are shown out (Table 2). The seasonal pattern of the N/P ratio showed a signicant increase of this parameter in October, which along with other processes could be related to a higher retranslocation of P than of N. This phenomenon is especially evident in young leaves, which become completely developed from spring to autumn and consequently under go no further growth.

Nutrient Seasonal Variation in Foxglove Table 2 Seasonal variation of N/P and Ca/Mg ratios in young and mature D. obscura leaves Ratio N/P Ca/Mg Leaf type Young Mature Young Mature May 8.38b 8.70b 2.89b 5.63a July 9.91b 11.49ab 4.10ab 5.06a October 13.34a 14.20a 4.83a 5.24a February 10.60b 12.11ab 5.09a 5.35a

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Values with the same letter in each row are not signicantly different according to Tukeys test (p < 0.05). Data are the means of the 10 populations studied.

At this time the P requirements for RNA are greatly reduced and nucleic-acid P can, therefore, be mobilized and retranslocated to sink organs, leading to higher plant-level N/P ratios (G sewell, 2004). This ratio has been used as diagnostic u indicator of N saturation and/or of the vegetative growth limitation by these nutrients, as well as to identify thresholds of nutrient limitation (G sewell, 2004). u The N/P ratios for the 10 populations of D. obscura studied are similar to the average ratios of terrestrial plant species (N/P = 1013/1) at their natural eld sites (Garten, 1976; G sewell, 2004); therefore, the plants studied seemingly u did not present a clear limitation for these nutrients. With regard to Ca/Mg ratio, the signicant increase observed in young leaves from May to February was due to Ca accumulation associated to leaf maturation. Nevertheless, in most populations, the values of this ratio in D. obscura leaves (young leaf = 2.15.6; mature leaf = 2.68.4) are lower than the Ca/Mg ratio of 7.7 reported for vascular and nonvascular plants (Garten, 1976). Of the different D. obscura populations, the lowest value of the Ca/Mg ratio was obtained in plants from Ayora, where it was 2.1 or 2.6 for young or mature leaf, respectively. This fact could be explained by a possibly greater Mg uptake by these plants, as the soil where this population grows showed the maximum EDTA-extractable Mg among all soils studied, and the lack of selectivity during Ca and Mg uptake and transport by plants is well known (White, 2001). Correlation Coefcient Between Elements The correlation coefcients between mineral elements in young and mature leaves are shown in Table 3. Young leaves presented a higher number of signicant correlations than did mature leaves; this fact could be related to a nutritional imbalance in the latter leaf type, as these leaves are pre-senescent. The maximum correlation coefcient observed in young leaves was obtained between N and P, which is comparable to the results reported in previous studies performed

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Table 3 Correlation coefcients between nutrients in young and mature D. obscura leaves Leaf type Young P K Ca Mg Fe Mn Cu Zn Mature P K Ca Mg Fe Mn Cu Zn

Ca

Mg

Fe

Mn

Cu

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0.858 0.756 0.772 0.517 0.644 0.513 n.s. n.s. n.s. 0.483 0.638 0.445 n.s. n.s. n.s. n.s. n.s. n.s. 0.502 0.465 0.379 n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.

n.s. 0.369 n.s. 0.354 n.s.

n.s. n.s. 0.313 n.s. n.s. n.s. n.s.

n.s. 0.363

0.375

0.338* 0.493 0.336 n.s. n.s. n.s. n.s. n.s. 0.345 n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.

n.s. n.s. n.s. n.s.

n.s. n.s. n.s.

n.s. 0.715

n.s.

p < 0.05; p < 0.01; n.s.: non-signicant correlation (n = 40).

with several species in natural environments (Meerts, 1997; Thompson et al., 1997). Thus, when a species is sampled at different eld sites, N and P are well correlated with each other because the same sites tend to be N- and P-rich, and because non-nutritional factors (e.g., shade, drought) have similar impacts on N and P (G sewell, 2004). The high correlation of these elements reects their u close association in plant biochemistry, particularly protein synthesis (Garten, 1976). On the other hand, Ca and Fe presented signicant negative correlations with N, P, and K, which corroborates the results previously outlined in relation to the accumulation process of Ca and Fe, and to the retranslocation of N, P, and K accompanying leaf aging. It has been reported that the signicant positive or negative correlations found between mineral elements in different plant organs may indicate synergistic or antagonistic interactions, respectively, of the nutrients analyzed (Maier and Chvyl, 2003; Saat i and Ya Mur, 2000). In this regard, our analyses showed c no signicant negative correlations between the micronutrients in D. obscura leaves (Table 3), which would lead to the conclusion that there are no antagonisms between the microelements measured. This result is supported by the fact that the soils of the 10 communities studied did not contain high concentrations of micronutrients (Roca-P rez et al., 2004a); therefore, the antagonism e

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phenomenon could not be provoked under the natural conditions of D. obscura populations. ACKNOWLEDGMENTS This research was supported by the Generalitat Valenciana (Project GV-DRN-12-147-96) and by a research grant to Luis Roca-P rez within the project e GV-CAPA00-03. REFERENCES

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