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[目的]筛选出简易的DNA提取方法及适合于所有浙江红山茶种质材料进行ISSR分析的有效引物。[方法]采用改良的CTAB法提取基因组DNA,参考其他山茶科植物的50个ISSR引物,对来自浙江、福建、江西、安徽10个居群中共20份浙江红山茶种质材料进行了PCR扩增。[结果]改进的CTAB法可简单、快速地提取到高纯度DNA产物;筛选出的20条引物多态性丰富、条带清晰且可重复性良好。共扩增出337条DNA谱带,其中281条为多态性带,占总扩增带数的83.4%,平均每个引物扩增出16.85条谱带。[结论]所筛选的20条引物可有效应用于浙江红山茶种质资源材料的ISSR分析。
[Objective] The research aimed to screen simple DNA extraction methods and effective primers for ISSR analysis of all Camellia japonica germplasm resources. [Method] The genomic DNA was extracted by modified CTAB method. According to 50 ISSR primers of other camellia plants, 20 accessions of Zhejiang red camellia from 10 populations of Zhejiang, Fujian, Jiangxi and Anhui were amplified by PCR increase. [Result] The improved CTAB method could extract the high purity DNA product easily and rapidly. The 20 selected primers had abundant polymorphism, clear bands and good reproducibility. A total of 337 DNA bands were amplified, of which 281 were polymorphic bands, accounting for 83.4% of the total bands amplified, with an average of 16.85 bands per primer. [Conclusion] The 20 primers screened could be effectively applied to the ISSR analysis of Zhejiang Hongshan tea germplasm resources.