睡莲种质资源遗传多样性分析及DNA指纹图谱构建

Genetic Diversity Analysis and DNA Fingerprinting Construction of Waterlily Germplasm Resources

利用ISSR分子标记技术对从国内外引种的46份睡莲种质资源进行遗传多样性分析,并对其中的原生种及原生变种构建DNA指纹图谱,旨在阐明其亲缘关系基础上,为睡莲分类鉴定、杂交育种、功能基因的挖掘和利用及图位克隆等提供理论依据。结果表明:从100条ISSR引物中筛选出10条多态性高、重复性好的引物,在46份睡莲种质资源中共扩增出281条带,平均每条引物扩增条带数为28.1条,多态性条带281条,多态性比例为100%;平均Shannon信息指数(I)为0.4197,平均Nei’s基因多样性指数(H)为0.2657,平均有效等位基因数(N)1.4139,表现出丰富的遗传多样性;遗传相似系数值在0.51~0.98之间,基于遗传相似系数建立了聚类树状图,揭示了46份睡莲种质资源的亲缘关系,并在相似系数水平为0.68时,可将所有供试材料聚为6类。对总计24个睡莲原生种及原生变种构建了DNA指纹图谱,依据图谱差异可鉴定不同的睡莲种质。

The genetic diversity of 46 waterlily germplasm resources introduced within China and abroad was analyzed by the ISSR molecular marker technology,and the DNA fingerprints of the original species and varieties were constructed in order to clarify the genetic relationship and provide a theoretical basis for the classification and identification of waterlily,hybrid breeding and functional gene mining,utilization and map-based cloning.The results showed that 10 primers with high polymorphism and good repeatability were screened out from 100 ISSR primers.A total of 281 bands were amplified from 46 waterlily germplasm resources,with an average of 28.1 bands per primer,281 polymorphic bands and 100%polymorphic ratio.The average Shannon information index(I)was 0.4197,the average Nei's gene diversity index(H)was 0.2657,and the average effective allele number(N)was 1.4139,which showed abundant genetic diversity.The genetic similarity coefficient was between 0.51 and 0.98.Based on the genetic similarity coefficient,a clustering tree was constructed to reveal the genetic relationship of the 46 waterlily germplasm resources.When the similarity coefficient was 0.68,all the tested materials could be clustered into six groups.DNA fingerprints were constructed for 24 original species and varieties of waterlily,and different waterlily germplasms could be identified according to the differences of DNA fingerprints.