利用RAPD分子标记快速鉴定3个奇楠种质

Rapid identification of 3 Chi-Nan germplasms by RAPD molecular Markers

利用随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)分子标记技术,分析3个奇楠种质(ChiNan germplasm)亲缘关系,并快速鉴定。通过提取基因组DNA,筛选适用于奇楠种质分析的RAPD引物,PCR扩增获得扩增条带,分析奇楠种质的遗传多样性,并利用人工绘制品种鉴别示意图方法(manual cultivar identification diagram,MCID)将奇楠种质区分开来。从120条RAPD分子标记引物中筛选到10条适合于奇楠种质分析的引物,利用这10条引物发现奇楠种质在物种水平上遗传多样性高,3个奇楠种质不同居群间的遗传一致性高、遗传距离小,在聚类图上各自聚为一支;根据引物S63、S18和S100扩增的多态性谱带构建奇楠种质的MCID,很好地区分了3个奇楠种质。3个奇楠种质均具有特异性强、一致性好、稳定性高的特点,RAPD分子标记技术的MCID可以有效快速地鉴定区分3个奇楠种质。

Rapid identification and genetic relationship of 3 Chi-Nan germplasms were analyzed by RAPD molecular markers. The genomic DNA was extracted,RAPD primers suitable for Chi-Nan germplasm analysis were screened,the amplified bands were obtained by PCR,and the genetic diversity of Chi-Nan germplasm was analyzed. Using MCID,3 Chi-Nan germplasms were gradually and completely distinguished by RAPD primers. 10 primers were screened from 120 RAPD molecular marker primers which were suitable for analysis of Chi-Nan germplasm. It was found that the genetic diversity of Chi-Nan germplasm was high at the species level by these 10 primers. The 3 germplasms showed high genetic consistency and small genetic distance among different populations,and were clustered into one branch respectively in phylogenetic tree. The MCID of Chi-Nan germplasm was constructed according to the pleomorphic bands amplified by primers S63,S18 and S100,which could separate 3 Chi-Nan germplasms from each other. 3 Chi-Nan germplasms have the characteristics of strong specificity,good consistency and high stability. The MCID based on RAPD molecular marker is able to identify and distinguish 3 Chi-Nan germplasms efficiently and rapidly.