Protein resources in wild barley,Hordeum spontaneum,in Israel: Predictive method by ecology and allozyme markers

Geographic variation in protein content of wild barley,Hordeum spontaneum, and the associations of protein content with ecological and allozyme markers were tested in an attempt to derive predictive guidelines for conservation and utilization in breeding programs. The study involved 195 genotypes of wild barley from 25 populations, 15 central and 10 marginal. These populations had been tested earlier for allozymic variation (Nevo & al. 1979 a, b). The results indicate that protein content varies both within, but particularly between populations. Notably, the 10 marginal populations exhibit high protein content but low kernel weight, as compared with the 15 central populations which displayed lower protein content but high kernel weight. Three variable combinations of climatic factors explain 40% of the variability in protein content among populations. Likewise, 3 variable combinations of allozyme allele frequencies explain a significant degree of spatial variance in protein content (R square = 0.63). — We conclude that natural populations of wild barley in Israel contain large amounts of yet untapped genes for protein content. These could be effectively screened and utilized for producing high protein cultivars of barley by following ecological and allozymic markers as predictive guidelines in screening natural populations of wild barley.     

Resistance of wild wheat to stripe rust: Predictive method by ecology and allozyme genotypes

From 114 accessions of wild emmer wheat from 11 sites in Israel, known for their allozymic variation (Nevo & al. 1982), individual genotypes were tested for resistance to one isolate of stripe rust both in the seedling stage in a growth chamber and in the adult plant stage in the field. The results indicate that resistance to stripe rust in seedlings and adults are significantly correlated (r_s = 0.40, p < 0.001). Genetic polymorphisms of resistance to stripe rust vary geographically and are predictable by climatic, as well as allozymic markers. Three variable combinations of rainfall, evaporation, and temperature explain significantly 0.40–0.53 of the spatial variance in disease resistance to stripe rust, suggesting the operation of natural selection. Several allozyme genotypes are significantly associated with disease resistance. We conclude that natural populations of wild emmer wheat in Israel contain large amounts of disease resistance genes. These populations could be effectively screened and then utilized by the phytopathologist for identifying resistant genotypes and producing new resistant cultivars.Patterns of Resistance of Wild Wheat to Pathogens in Israel II.     

Photosynthetic performance in wild emmer wheat,Triticum dicoccoides: ecological and genetic predictability

Summary In a twin study, we have shown that wild emmer wheat, Triticum dicoccoides, the progenitor of all cultivated wheats, harbours important genetic variation (Vg) in photosynthetic characteristics. This Vg resides within and between populations and ecogeographical regions in Israel, which is the center of origin and diversity of wild emmer wheat. Here we analyzed, by univariate and multivariate methods, the significant differentiation of variation in photosynthetic characteristics of 107 genotypes from 27 populations of wild emmer in Israel, distributed in three ecogeographical regions including central, xeric (northern cold and eastern warm) marginal, and mesic (western) marginal populations. The highest photosynthetic efficiency was displayed by populations of the xeric marginal region, but most variation for photosynthetic capacity occurs between accessions within ecogeographical regions and populations. Genotypes and populations of T. dicoccoides having high photosynthetic capacity can be identified by climatic factors and isozyme markers. The identification by genetic markers, if substantiated by testcrosses, can facilitate the maximization of conservation, in situ or ex situ, and utilization of these photosynthetic genetic resources for improvement of hexaploid wheat (T. aestivum).     

Patterns of resistance of Israeli wild emmer wheat to pathogens I. Predictive method by ecology and allozyme genotypes for powdery mildew and leaf rust

The association of ecological factors and allozyme markers with genotypes of tetraploid wild emmer wheat,Triticum dicoccoides, varying in resistance to four cultures of the pathogenErysiphe graminis tritici, and to one culture ofPuccinia recondita tritici, which incite the diseases powdery mildew and leaf rust respectively, were explored theoretically and practically. The study involved 233 accessions comprising 10 populations representing the ecological range ofT. diococcoides in Israel. Our results indicate that genetic polymorphism for resistance to both pathogens is structured geographically, and is predictable by climatic as well as allozymmc markers. Three variable combinations of water factors and temperature differentials significantly explain 0.27 and 0.14 of the spatial variance for resistance to powdery mildew and leaf rust, respectively, suggesting the involvement of natural selection. Several allozyme genotypes, singly, or in combination, are significantly associated with disease resistance. We conclude thatT. dicoccoides populations in Israel, which grow in the center of diversity of the species, contain large amounts of unexploited disease resistant genotypes. The populations could be effectively screened and utilized for producing resistant cultivars by means of ecological factors and allozyme markers as predictive guidelines.     

Microsatellite polymorphism in natural populations of wild emmer wheat,Triticum dicoccoides,in Israel

Diversity in 20 microsatellite loci of wild emmer wheat, Triticum dicoccoides, was examined in 15 populations (135 genotypes) representing a wide range of ecological conditions of soil, temperature, and water availability, in Israel and Turkey. An extensive amount of diversity at microsatellite loci was observed despite the predominantly selfing nature of this plant species. The 20 Gatersleben wheat microsatellites (GWM), representing 13 chromosomes of genomes A and B of wheat, revealed a total of 364 alleles, with an average of 18 alleles per GWM marker (range: 5–26). The proportion of polymorphic loci per population averaged 0.90 (range: 0.45– 1.00); genic diversity, He, averaged 0.50 (range 0.094– 0.736); and Shannon’s information index averaged 0.84 (range 0.166–1.307). The coefficients of genetic distance between populations were high and averaged D=1.862 (range 0.876–3.320), an indication of sharp genetic divergence over short distances. Interpopulation genetic distances showed no association with geographic distance between the population sites of origin, which ruled out a simple isolation by distance model. Genetic dissimilarity values between genotypes were used to produce a dendrogram of the relationships among wild wheat populations by the unweighted pair-group method with arithmetic averages (UPGMA). The results showed that all the wild emmer wheat populations could be distinguished. Microsatellite analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides, originating from diverse ecogeographical sites in Israel and Turkey, with 88% of the 135 genotypes correctly classified into sites of origin by discriminant analysis. Our present microsatellite results are non-random and in agreement with the previously obtained allozyme and RAPD patterns, although the genetic-diversity values obtained with microsatellites are much higher. Significant correlates of microsatellite markers with various climatic and soil factors suggest that, as in allozymes and RAPDs, natural selection causes adaptive microsatellite ecogeographical differentiation, not only in coding, but most importantly in non-coding genomic regions. Hence, the concept of ”junk DNA” needs to be replaced by at least partly regulatory DNA. The obtained results suggest that microsatellite markers are useful for the estimation of genetic diversity in natural populations of T. dicoccoides and for the tagging of agronomically important traits derived from wild emmer wheat. Received: 27 February 2001 / Accepted: 22 March 2001     

Spatiotemporal allozyme divergence caused by aridity stress in a natural population of wild wheat, Triticum dicoccoides, at the Ammiad microsite, Israel

Spatiotemporal diversity at 35 allozyme loci was assayed over 6 years in 1,207 individuals of wild emmer wheat (Triticum dicoccoides)from a microgeographic microsite, Ammiad, north Israel. This analysis used new methods and two additional sample sets (1988 and 1993) and previous allozymic data (1984–1987). This microsite includes four major habitats (North-facing slope, Valley, Ridge, and Karst) that show topographic and ecological heterogeneity. Significant temporal and spatial variations in allele frequencies and levels of genetic diversity were detected in the four subpopulations. Significant associations were observed among allele frequencies and gene diversities at different loci, indicating that many allele frequencies change over time in the same or opposite directions. Multiple regression analysis showed that variation in soil-water content and rainfall distribution in the growing season significantly affected 10 allele frequencies, numbers of alleles at 8 loci, and gene diversity at 4 loci. Random genetic drift and hitchhiking models may not explain such locus-specific spatiotemporal divergence and strong allelic correlation or locus correlation as well as the functional importance of allozymes. Natural ecological selection, presumably through water stress, might be an important force adaptively directing spatiotemporal allozyme diversity and divergence in wild emmer wheat at the Ammiad microsite. Received: 3 July 2000 / Accepted: 1August 2000     

Correlation of ecological factors and allozymic variations with resistance toErysiphe graminis hordei inHordeum spontaneum in Israel: Patterns and application

The association of ecological factors and allozymic markers of wild barley,Hordeum spontaneum, with genotypes varying in resistance to 3 cultures of the pathogenErysiphe graminis hordei, which incites the disease powdery mildew of barley, were explored theoretically and practically. The study involved 275 accessions comprising 16 populations largely representing the ecological range ofH. spontaneum in Israel. From earlier studies of allozymic variation and disease resistance it now becomes apparent that genetic polymorphisms for resistance toE. graminis hordei are structured geographically, and are predictable by climatic as well as allozymic variables. Three-variable combinations of temperature and water factors explain significantly 0.32 of the spatial variance in disease resistance between localities. Also, several allozyme genotypes, singly or in combination, are significantly associated with disease resistance. A high correlation was found between the standard deviation of infection types of the culture of the pathogen from Israel, and allozymic polymorphism,P (r_s = 0.86, p < 0.001). Consequently, the IsraelH. spontaneum populations, growing in the center of diversity of the species, contain large amounts of unexploited disease resistance polymorphism. These could be effectively screened and utilized for producing resistant barley varieties by using ecological factors and allozymic variants as guidelines.     

RAPD polymorphism of wild emmer wheat populations, Triticum dicoccoides, in Israel

 Genetic diversity in random amplified polymorphic DNAs (RAPDs) was studied in 110 genotypes of the tetraploid wild progenitor of wheat, Triticum dicoccoides, from 11 populations sampled in Israel and Turkey. Our results show high level of diversity of RAPD markers in wild wheat populations in Israel. The ten primers used in this study amplified 59 scorable RAPD loci of which 48 (81.4%) were polymorphic and 11 monomorphic. RAPD analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides originating from diverse ecogeographical sites in Israel and Turkey, with 95.5% of the 100 genotypes correctly classified into sites of origin by discriminant analysis based on RAPD genotyping. However, interpopulation genetic distances showed no association with geographic distance between the population sites of origin, negating a simple isolation by distance model. Spatial autocorrelation of RAPD frequencies suggests that migration is not influential. Our present RAPD results are non-random and in agreement with the previously obtained allozyme patterns, although the genetic diversity values obtained with RAPDs are much higher than the allozyme values. Significant correlates of RAPD markers with various climatic and soil factors suggest that, as in the case of allozymes, natural selection causes adaptive RAPD ecogeographical differentiation. The results obtained suggest that RAPD markers are useful for the estimation of genetic diversity in wild material of T. dicoccoides and the identification of suitable parents for the development of mapping populations for the tagging of agronomically important traits derived from T. dicoccoides. Received: 13 July 1998 / Accepted: 13 August 1998     

Natural selection of allozyme polymorphisms: a microgeographic climatic differentiation in wild emmer wheat (Triticum dicoccoides)

Summary Allozymic variation in proteins encoded by 48 loci was analyzed electrophoretically in 1984 and 1985 in 137 individual plants of wild emmer wheat, Triticum dicoccoides, from a microsite in Yehudiyya, northeast of the Lake of Galilee, Israel. The test involved two climatic microniches in the open Tabor oak forest (1) sunny between trees and (2) shady under trees' canopies. Significant genetic differentiation at single-, two- and multilocus structures was found between neighboring climatic niches, which were only separated by a few meters. Our results suggest that allozyme polymorphisms in wild emmer wheat are partly adaptive, and differentiate primarily at the multilocus level by climatic factors presumably related to aridity stress.     

The structure of wild and domesticated emmer wheat populations,gene flow between them,and the site of emmer domestication

The domestication of emmer wheat (Triticum turgidum spp. dicoccoides, genomes BBAA) was one of the key events during the emergence of agriculture in southwestern Asia, and was a prerequisite for the evolution of durum and common wheat. Single- and multilocus genotypes based on restriction fragment length polymorphism at 131 loci were analyzed to describe the structure of populations of wild and domesticated emmer and to generate a picture of emmer domestication and its subsequent diffusion across Asia, Europe and Africa. Wild emmer consists of two populations, southern and northern, each further subdivided. Domesticated emmer mirrors the geographic subdivision of wild emmer into the northern and southern populations and also shows an additional structure in both regions. Gene flow between wild and domesticated emmer occurred across the entire area of wild emmer distribution. Emmer was likely domesticated in the Diyarbakir region in southeastern Turkey, which was followed by subsequent hybridization and introgression from wild to domesticated emmer in southern Levant. A less likely scenario is that emmer was domesticated independently in the Diyarbakir region and southern Levant, and the Levantine genepool was absorbed into the genepool of domesticated emmer diffusing from southeastern Turkey. Durum wheat is closely related to domesticated emmer in the eastern Mediterranean and likely originated there. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Natural selection of allozyme polymorphisms: a microgeographical differentiation by edaphic,topographical, and temporal factors in wild emmer wheat (Triticum dicoccoides)

Summary Allozymic variation in proteins encoded by 47 loci was analyzed electrophoretically in 1983/4 and 1984/5 in 356 individual plants of wild emmer wheat, Triticum dicoccoides, from a microsite at Tabigha, north of the Sea of Galilee, Israel. Each year the test involved two 100-meter transects, each equally subdivided into basalt and terra rossa soil types, and comparisons were based on 16 common polymorphic loci. Significant genetic differentiation, genetic phase disequilibria, and genome organization according to soil type were found over very short distances. Our results suggest that allozyme polymorphisms in wild emmer wheat are partly adaptive, and that they differentiate at both single and multilocus structures primarily from environmental stress of such ecological factors as soil type, topography, and temporal changes, probably through aridity stress.     

Geographical variation in heading traits in wild emmer wheat, Triticum dicoccoides. I. Variation in vernalization response and ecological differentiation

 Geographical variation in vernalization response and narrow-sense earliness was investigated for accessions of wild emmer wheat, Triticum dicoccoides, collected in Israel. Wide variation between and within populations was observed in both characters. The analysis of vernalization response showed that 2 accessions from Tabigha were of a strong spring growth habit, and thus wild emmer wheat was classified into four types, i.e., strongly spring type, moderately spring type, moderately winter type, and strongly winter type, according to their vernalization response. Whereas winter types were frequently found in most populations except that of Tabigha, the distribution of spring types was sporadic and restricted to warmer areas. It was thus suggested that spring type in T. dicoccoides might have evolved from a winter prototype as an adaptation to warmer conditions. Within moderately winter and moderately spring types, quantitative differences in vernalization response, measured as Dof₇₀/Dof₂₀ and Dof₂₀/Dof₀, were observed between populations. Inter- and intra-population variation in vernalization response could be explained to some extent by the difference in growing conditions at each habitat. It was clearly indicated that environmental heterogeneity caused ecogenetic differentiation in wild emmer wheat in Israel. Wild emmer wheat also varied considerably for narrow-sense earliness, ranging from 32.9 days to 69.5 days among accessions. However, it was difficult to explain its geographical variation simply by a linear relationship with environmental factors, and a nonlinear relationship and/or unknown microgeographic heterogeneity may be responsible. Received: 18 March 1996/Accepted: 13 December 1996     

Wheat storage proteins: diversity of HMW glutenin subunits in wild emmer from Israel

Summary The diversity of HMW glutenin subunits in the tetraploid wild progenitor of wheat, Triticum turgidum var. dicoccoides was studied electrophoretically in 231 individuals representing 11 populations of wild emmer from Israel. The results show that (a) The two HMW glutenin loci, Glu-A1 and Glu-B1, are rich in variation, having 11 and 15 alleles, respectively, (b) Genetic variation in HMW glutenin subunits is often severely restricted in individual populations, supporting an island population genetic model, (c) Significant correlations were found between glutenin diversity and the frequencies of specific glutenin alleles and physical (climate and soil) and biotic (vegetation) variables. Our results suggest that: (a) at least part of the glutenin polymorphisms in wild emmer can be accounted for by environmental factors and (b) the endosperm of wild emmer contains many allelic variants of glutenin storage proteins that are not present in bread wheat and could be utilized in breeding varieties with improved bread-making qualities.     

Genetic diversity for drought resistance in wild emmer wheat and its ecogeographical associations

Wild emmer wheat (Triticum turgidum spp. dicoccoides (Körn.) Thell.), the tetraploid progenitor of cultivated wheat, is a potential source for various agronomical traits, including drought resistance. The objectives of this study were to characterize (1) the genetic diversity for drought resistance in wild emmer wheat, and (2) the relationship between drought responses of the wild emmer germplasm and the ecogeographical parameters of its collection sites. A total of 110 wild emmer accessions consisting of 25 populations and three control durum wheat cultivars were examined under two irrigation regimes, well-watered (’wet’) and water-limited (’dry’). Wide genetic diversity was found both between and within the wild emmer populations in most variables under each treatment. A considerable number of the wild emmer accessions exhibited an advantage in productivity (spike and total dry matter) over their cultivated counterparts. Most wild emmer wheat accessions exhibited a greater carbon isotope ratio (δ¹³C, indicating higher water-use efficiency) under the dry treatment and higher plasticity of δ¹³C relative to the cultivated controls, which may have contributed to the drought adaptations in the former. The most outstanding drought-tolerance capacity (in term of productivity under the dry treatment and susceptibility indices) was detected in wild emmer populations originated from hot dry locations. The results suggest that wild emmer has the potential to improve drought resistance in cultivated wheat.     

Genetic resources of wild barley in the Near East: structure, evolution and application in breeding

Genetic diversity and structure of populations of the wild progenitor of barley, Hordeum spontaneum, from three countries, Israel, Turkey and Iran, in the Near East Fertile Crescent, are compared and contrasted. The analysis is based on electrophoretically discernible allozymic variation in proteins encoded by 27 shared loci in 2125 individuals representing 52 populations of wild barley. The results indicate that: (a) H. spontaneum in the Near East Fertile Crescent is very variable genetically; (b) genetic differentiation of populations includes some clinal but primarily regional and local patterns often displaying sharp geographic differentiation over short distances; (c) the average relative genetic differentiation (G_st) was 54% within populations, 39% among populations, and 8% between the three countries; (d) allele distribution is characterized by a high proportion of unique alleles (51%), and a high proportion of common alleles that are distributed either locally or sporadically; (e) discriminant analysis by allele frequencies successfully clustered wild barley of each of the three countries (96% correct classification); (f) a substantial portion of the patterns of allozyme variation in the wild gene pool is significantly correlated with the environment and is predictable ecologically, chiefly by a combination of humidity and temperature variables; (g) natural populations of wild barley are, on the average, more variable than two composite crosses and land races of cultivated barley. The spatial patterns and environmental correlates and predictors of genetic variation of H, spontaneum in the Fertile Crescent indicate that genetic variation in wild barley populations is not only rich but at least partly adaptive and predictable by ecology and allozyme markers. Consequently, conservation and utilization programmes should optimize sampling strategies by following the ecological-genetic factors and allozyme markers as effectively predictive guidelines.     

Genetic diversity of wild emmer wheat in Israel and Turkey

Summary Allozyme variation in the tetraploid wild emmer wheat, Triticum dicoccoides, the progenitor of all cultivated wheats, was studied for the proteins encoded by 42 gene loci in 1815 plants representing 37 populations - 33 from Israel and 4 from Turkey - sampled in 33 localities from 1979 to 1987. The results showed that: (a) 6 loci (14%) were monomorphic in all populations, 15 loci (36%) were locally polymorphic, and 21 loci (50%) were regionally polymorphic. These results are similar to those obtained earlier on 12 Israeli populations. All polymorphic loci (except 4) displayed high local levels of polymorphism (>/ 10%). (b) The mean number of alleles per locus, A, was 1.252 (range: 1.050–1.634); the proportion of polymorphic loci per population averaged 0.220 (range: 0.050–0.415); genic diversity, He, averaged 0.059 (range: 0.002–0.119). (c) Altogether there were 119 alleles at the 42 putative loci tested, 114 of these in Israel, (d) Genetic differentiation was primarily regional and local, not clinal; 70% of the variant alleles were common (>/ 10%) and not widespread, but rather localized or sporadic, displaying an archipelago population genetics and ecology structure. The coefficients of genetic distance between populations were high and averaged D = 0.134; range: 0.018–0.297, an indication of sharp genetic differentiation over short distances, (e) Discriminant analyses differentiated Israeli from Turkish populations, and within Israel, between central and 3 marginal regions, as well as between different soil-type populations, (f) Allozymic variation comprised 40% within and 60% between populations, (g) Gametic phase disequilibria were abundant, their number being positively correlated (r_s = 0.60, P<0.01) with the humidity, (h) Multilocus organization was substantive, also positively correlated with humidity, (i) Allozyme diversity, overall and at single loci, was significantly correlated with, and partly predictable by, climatic and edaphic factors, (j) The distrubition of the significant positive and negative values and the absence of autocorrelations in the correlogram revealed no similar geographic patterns across loci, eliminating migration as a prime factor of population genetic differentiation. These results suggest: (I) during the evolutionary history of wild emmer, diversifying natural selection, through climatic and edaphic factors, was a major agent of genetic structure and differentiation at both the single and multilocus levels; (II) wild emmer harbors large amounts of genetic diversity exploitable as genetic markers in sampling and abundant genetic resources utilizable for wheat improvement.     

Genetic diversity of photosynthetic characters in native populations of Triticum dicoccoides

Wild emmer wheat (Triticum dicoccoides Korn) has shown wide genetic diversity for disease resistance and morpho-physiological traits of economic importance. Our objectives were to test for genetic variation (V_G) in photosynthetic characteristics residing within and between native populations sampled from three ecogeographical regions of Israel, and to identify potential sources of high photosynthetic efficiency for future wheat improvement. Accessions sampled in the center of wild emmer distribution (upper Jordan Valley) in a relatively narrow geographical range showed the greatest diversity in CO₂-assimilation rate per unit leaf area (A) or per unit chlorophyll (A/Chl). Genetic variation was absent for internal CO₂ concentration (Ci) and water-use efficiency (WUE) and generally lacking for stomatal conductance (gs). Leaf area, although quite variable, was not a significant cofactor in assessing genetic potential for photosynthesis. Accessions within a given population showed 10-times more variation in A and A/Chl than populations sampled from different locations in a region. Accessions with the highest photosynthetic efficiency were derived from upland steppic populations located in marginal habitats extending southward into Israel. Some accessions having high photosynthetic capacity (A=32 mol m^-2 s^-1) with no significant reduction in leaf size constitute a potentially valuable genetic resource yet untapped for genetic improvement of hexaploid (T. aestivum L.) wheat.Abbreviations LA total leaf area - V_G total genetic variance - PPFD photosynthetic photon flux density - WUE water-use efficiency     

Genetic diversity for grain nutrients in wild emmer wheat: potential for wheat improvement

Background and Aims

Micronutrient malnutrition, particularly zinc and iron deficiency, afflicts over three billion people worldwide due to low dietary intake. In the current study, wild emmer wheat (Triticum turgidum ssp. dicoccoides), the progenitor of domesticated wheat, was tested for (1) genetic diversity in grain nutrient concentrations, (2) associations among grain nutrients and their relationships with plant productivity, and (3) the association of grain nutrients with the eco-geographical origin of wild emmer accessions.

Methods

A total of 154 genotypes, including wild emmer accessions from across the Near Eastern Fertile Crescent and diverse wheat cultivars, were characterized in this 2-year field study for grain protein, micronutrient (zinc, iron, copper and manganese) and macronutrient (calcium, magnesium, potassium, phosphorus and sulphur) concentrations.

Key Results

Wide genetic diversity was found among the wild emmer accessions for all grain nutrients. The concentrations of grain zinc, iron and protein in wild accessions were about two-fold greater than in the domesticated genotypes. Concentrations of these compounds were positively correlated with one another, with no clear association with plant productivity, suggesting that all three nutrients can be improved concurrently with no yield penalty. A subset of 12 populations revealed significant genetic variation between and within populations for all minerals. Association between soil characteristics at the site of collection and grain nutrient concentrations showed negative associations between soil clay content and grain protein and between soil-extractable zinc and grain zinc, the latter suggesting that the greatest potential for grain nutrient minerals lies in populations from micronutrient-deficient soils.

Conclusions

Wild emmer wheat germplasm offers unique opportunities to exploit favourable alleles for grain nutrient properties that were excluded from the domesticated wheat gene pool.     

Microclimatic stress and adaptive DNA differentiation in wild emmer wheat, Triticum dicoccoides

 Genetic diversity was examined by RAPD-PCR analysis in 118 registered individuals of wild emmer wheat, Triticum dicoccoides, from a microsite at Yehudiyya, northeast of the Sea of Galilee, Israel. The test involved two climatic microniches in the open oak-park forest of Quercus ithaburensis: (1) sunny between trees and (2) shady under the trees’ canopies. Comparisons were based on 97 loci amplified by 20 oligonucleotide primers. Significant genetic differentiations were found at single-, two- and multilocus structures between the neighbouring shady and sunny niches. These DNA polymorphisms appear to be associated with microclimatic stresses. The pronounced niche-effect on the significance of linkage disequilibrium and niche-specific linkage disequilibrium may suggest that natural selection directed the two-locus associations. The structure of the multilocus associations also mainly results from natural selection, and not by chance from population subdivision, or founder effects. These findings are largely parallel to the previous allozymic results at single-locus and multilocus levels. Both the DNA and the allozymic results suggest that microclimatic selection appears to play an important role in DNA differentiation as well as in protein polymophism. Received: 30 October 1998 / Accepted: 2 November 1998