野生大麻种质资源表型及其RAPD遗传多样性分析

Genetic Diversity Analysis of Wild Cannabis in China Based on Morphological Characters and RAPD Markers

以中国12份野生大麻种质及4个对照栽培品种为研究对象,通过田间栽培试验,调查叶长、叶宽和叶柄等11个表型性状,并采用CTAB法提取大麻基因组DNA,分析了其表型性状及RAPD标记位点的多态性,应用Farthest neighbor和UPGMA方法分别构建了表型及RAPD聚类图。结果表明,野生大麻表型变异非常丰富,11个表型在不同种质资源间的差异性均达到了极显著水平(P<0.001),其中变异最大的为千粒重,变异最小的为有效分枝数;14条RAPD引物共扩增出106条带,其中79条为多态性条带,多态性比率为74.52%。基于表型聚类分析,16份大麻种质资源分为3个类群,第1类群包括全部12份野生大麻种质资源,且根据高纬和中低纬分为2个明显的分支,另外2个类群仅包括3份栽培大麻;基于RAPD聚类分析,16份大麻种质资源同样分为3个类群,总体上呈现地域性分布,但野生大麻和栽培大麻并未区分开,云南、新疆的种质资源聚为一类,东北、华北的种质资源聚为一类,西藏种质资源单独聚为一类。研究表明,我国野生大麻种质资源具有复杂的遗传多样性,应该结合表型和遗传位点综合分析。研究结果为野生大麻的利用和工业大麻品种改良提供参考依据。

Wild Cannabis germplasms is not only the natural gene bank of Cannabis research and utilization, but also the important basis of hemp breeding. In this article, genetic diversity of 12 wild Cannabis resources and 4 cul- tivars were assessed by the methods of phenotypic characters and RAPD markers, and the cluster of phenotypic traits and RAPD markers were constructed by Farthest-neighbor method and UPGMA method respectively. The re- suits showed the genetic diversity in wild cannabis was abundant, and the difference of 11 Morphological characters in all germplasms was significant （P 〈 0. 001 ）. The thousand-grain weight had the highest coefficient of variation, while the effective number of branches had the smallest one. After the amplification with 13 RAPD primers, a total of 106 fragments were detected, among which, 79 fragments were polymorphic, accounting for 74.52 %. The whole germplasms could be classified into three groups based on morphological cluster. The first group was com- posed of 12 wild germplasms which further divided into two subgroups based on the degree of latitude, while the other two groups contain only 3 cuhivars. RAPD cluster analysis showed that the whole germplasms were also classi-fled into 3 groups clustered with the characteristic of regionalism. Germplasms in Yunnan and Xinjiang were in one group, germplasms in northeast and north China were in the second group, while germplasm in Xizang was alone in another cluster, the wild and cultivars were not distinguished obviously. The results suggest that the genetic diversi- ty of Cannabis in china is complicated.