怀区地黄遗传多样性的ISSR鉴定

ISSR identification of genetic diversity of Rehmannia glutinosa in Huai zone

目的利用ISSR标记技术对怀区地黄的8个品种和2个茎尖培养脱毒系进行了遗传多样性分析。方法用44条ISSR引物进行初选,然后用其中适合的10个ISSR引物进行ISSR分析。结果10条ISSR引物共扩增出110条带。基于这些条带,用SPSS10.0软件分析,建立了遗传Jaccard相关系数矩阵,构建了分子树状图,将怀区地黄的8个品种和2个组培系分为2类:其中一类群含有6个材料,包括组培85.5、大田85.5、组培9302、大田9302、金状元和金白地黄;另一类群含有4个材料,包括北京1号、大红袍、地黄9104和野生地黄。而且主成分分析结果支持上述的聚类分析结果;用POPGENE32软件分析知,多态性指数为0.3775,有效等位基因数为1.4037,多态性比率为71.82%;用一个ISSR6引物建立了10个供试地黄样品的DNA指纹图谱并且能将其彼此区分出来。结论ISSR标记适合于构建怀区地黄的DNA指纹图谱、品种鉴定和遗传多样性分析。

Objective In order to characterize eight cultivars and two virus-free lines micro-propagated by tip tissue culture of Rehmannia glutinosa in Huai zone and to assess their genetic diversities by inter-simple sequence repeat (ISSR) technique. Methods Ten appropriate ISSR primers were selected from a total of 44 ISSR ones for ISSR PCR amplification. Results The ten primers could amplify one hundred and ten bands. Based on them, a Jaccard's genetic similarity matrix and a dendrogram for these ten cultivars were established using SPSS 10.0 software. In this dendrogram, they could be divided into two groups: group 1 contained six individuals, such as Zupei 85.5, Datian 85.5, Zupei 9302, Datian 9302, Jinzhuangyuan, and Jinbai Dihuang; group 2 consisted of four ones such as Beijing No.1, Dahongpao, Dihuang 9104, and wild Dihuang. Furthermore, principal coordinate analysis (PCA) supported the above cluster analysis; Shannon's information index (Ⅰ) is 0.377 5, effective number of alleles (Ne) is 1.403 7 , the percentage of polymorphic loci is 71.82% by means of POPGENE32 software; a DNA fingerprint was developed with a single primer, ISSR6, in which each of ten tested individuals had its unique fingerprint pattern and was distinguished from each other. Conclusion The results reveal that ISSR method is suitable for DNA fingerprinting, identification, and genetic diversity analysis of R. glutinosa in Huai zone