菜豆种质资源的ISSR分析及特异性标记的筛选

Analysis of Common Bean Germplasms Using ISSR and Screening of Specific Markers

采用ISSR分子标记技术对吉林省各科研院所近年审定和主栽的36份菜豆种质的遗传多样性进行研究。从100个引物中筛选出可扩增出清晰稳定条带、重复性好的20个引物对36个菜豆品种的基因组DNA进行PCR扩增。采用NTSYSpc2．10软件计算遗传相似系数，利用UPGMA方法进行聚类分析。结果表明：20个引物共扩增出87条带，其中多态性谱带76条，多态性比率为87．36％，多态性较高。36个品种间遗传相似系数在0．65～0．89之间。利用UPGMA法可将供试菜豆分为4大类，聚类结果显示，菜豆种质资源间的亲缘关系与来源地的相关性较为紧密，20号菜豆与其他品种的遗传距离相对较远。引物840为20号品种的特异性标记，在1100bp处多出一条谱带；引物861为27号品种的特异性标记，在1000bp处有一条谱带缺失；引物846为31号品种的特异性标记，在1050bp处有一条谱带缺失。3个引物针对20、27、31号品种的特异性标记可将3个品种快速从所有供试的36个菜豆品种中鉴别出来。

ISSR marker was used to identify the genetic diversity and relationship of 36 common bean cuhivars collected from the scientific research institutes in Jilin Province. 20 primers were screened out from the 100 ISSR primers. The genetic similarity coefficients were calculated with NTSYSpc2.10 and the cluster analysis was conducted by UPGMA. The results showed that 87 DNA fragments were amplified by PCR, a- mong which 76 ones were polymorphic and the polymorphism frequency was 87.36%. The results showed that there was relatively high genetic diversity among the accessions. The values of genetic similarity coefficients among the accessions were between 0.65 and 0.89. The 36 common bean accessions were divided into four groups by UPGMA, the cluster analysis conducted with UPGMA showed that the clustering results were closely correlated with the source of common bean and the genetic distance between cultivar No.20 and other cultivars was relatively remote. The specific makers of cuhivar No.20 based on primer 840 amplified an extra band in 1100bp, the specific makers of cultivar No.27 based on primer 861 amplified an absent band in 1000bp, and the specific makers of cuhivar No.30 based on primer 846 amplified an absent band in 1050bp. 3 specific makers could quickly identify cuhivar no.20, No.27 and No.31 from all the 36 common bean varieties.