利用ISSR标记研究野大豆居群内遗传变异及其取样策略

为了有效地保护野大豆(Glycine soja Sieb.et Zucc.)并制定合理的居群取样策略，对上海江湾机场的一个天然野大豆居群进行了100个单株(个体)的随机取样，并用ISSR分子标记对其进行了遗传多样性分析。利用筛选出的15条ISSR引物在这个居群中检测到较高的遗传变异，样本内个体间的相似系数变化在0．17～0．89之间。居群内平均每个位点的平均预期杂合度(He)为0．1714，香农指数(I)为0．2714。PCA分析显示，江湾野大豆居群内的遗传变异不是呈均匀分布，而是呈丛状分布。该野大豆居群遗传多样性和样本内个体数量间的相关性分析显示：在个体数少于40的情况下，遗传多样性随个体数的增加而迅速增加：当样本中的个体数大于40时，遗传多样性的增加减慢并很快趋于饱和。研究表明：对野大豆居群进行异地保护时，对各居群的采样植株数不应当低于35～45；在居群内采样时，所采集的个体之间最好相隔一定的空间距离。

Sampling Strategy Within a Wild Soybean Population Based on Its Genetic Variation Detected by ISSR Markers

In order to determine an appropriate sampling strategy for the effective conservation of wild soybean (Glycine soja Sieb. et Zucc.) in China, a natural population from Jiangwan Airport in Shanghai was studied for its genetic diversity through the inter-simple sequence repeat (ISSR) marker analysis of a sample set consisting of 100 randomly collected individuals. A relatively large genetic diversity was detected among the samples based on estimation of DNA products amplified from 15 selected ISSR primers, with the similarity coefficient varying from 0.17 to 0.89. The mean expected heterozygosity (He) was 0.171 4 per locus, and Shannon index (1) was 0.271 4. The Principal Coordinate Analysis (PCA) further indicated that genetic diversity of the Jiangwan wild soybean population was not evenly distributed, instead, was presented by a mosaic or clustered distribution pattern. Correlation study between genetic diversity and number of samples demonstrated that genetic diversity increased dramatically with the increase of number of samples within 40 individuals, but the increase became slow and rapidly reached a plateau when more than 40 individuals were included in the analysis. It is concluded that (i) a sample set of approximately 35-45 individuals should be included to represent possibly high genetic diversity when conservation of a wild soybean population ex situ is undertaken; and (ii) collection of wild soybean samples should be spread out as wide as possible within a population, and a certain distance should be kept as intervals among individuals for sampling.