11种荆芥种质资源ISSR分析

利用ISSR分子标记技术对不同地区的11种荆芥种质资源进行分析,以此来探讨我国荆芥种质资源的遗传多样性。以11种荆芥幼苗为材料,CTAB法提取其基因组DNA;采用正交试验优化ISSR技术体系影响因素;利用100条ISSR引物进行ISSR扩增,最后计算供试材料之间的遗传距离并进行聚类分析。结果表明,最优ISSR-PCR体系,在25 μL体系中,模板DNA 50 ng,引物0.75 μmol·L^-1,Taq DNA聚合酶1 U;100条引物中55条引物可扩增出结果,共扩增出458条条带,平均每个引物扩增出8.3条,其中246条为多态性带,多态性比率为53.71%;11个荆芥种质资源的遗传距离在0.208 3~0.709 1;在遗传距离为0.50处,可将供试的11种荆芥材料分为2大类;在遗传距离为0.40处,可将供试的11种荆芥材料分为5大类,研究结果表明我国荆芥种质资源遗传多样性较低。

Analysis of germplasm resources in 11 varieties Schizonepeta tenuifolia Briq. by ISSR

The aim of the study was to analyze the genetic diversity of 11 Schizonepeta tenuifolia Briq.using ISSR, which would investigate the genetic diversity of Schizonepeta tenuifolia Briq. germplasm resources in China. With 11 kinds of Schizonepeta tenuifolia Briq. seedlings as materials, genomic DNA was extracted using the CTAB method. An orthogonal design was used to optimize the ISSR-PCR amplification system. 100 ISSR primers were used in the ISSR amplification. At last it counted the genetic distance between the tested materials and finally the cluster analysis was done. The results showed that a suitable ISSR reaction system was constructed with 25 μL reaction system containing 50 ng DNA, 0.75 μmol·L^-1 primer and 1 U Taq DNA polymerase. There were 55 primers amplified from the 100 primers, 458 bands were amplified, 8.3 bands were amplified from each primer, of which 246 bands were polymorphic, and the polymorphic rate was 53.71%. The genetic distances of 11 Schizonepeta tenuifolia Briq. germplasm resources were between 0.2083-0.7091. When the genetic distance was 0.50, 11 kinds of materials could be divided into 2 categories. When the genetic distance was 0.40, 11 kinds of materials could be divided into 5 categories. In conclusion, the genetic diversity of germplasm resources of Schizonepeta tenuifolia Briq. was lower in China.