浙江红山茶总DNA提取及ISSR-PCR引物筛选

DNA Extraction and ISSR Primer Screening of Camellia chekiangoleosa

[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 aim was to screen out simple methods of DNA extraction and effective ISSR primers suitable to all germplasm materials of Camellia chekiangoleosa.[Method] The modified CTAB method was used in the extraction of the genomic DNA,50 ISSR primers from other Camellia plants were used in the ISSR-PCR amplification for 20 germplasm materials from 10 populations in Fujian,Zhejiang,Jiangxi and Anhui Province.[Result] Pure DNA could be obtained rapidly by using the improved CTAB method,and the 20 selected effective primers had rich polymorphism,clear bands and good repeatability.337 DNA bands were obtained,of which 281 bands were polymorphic,accounting for 83.4% of the total amplified bands.And 16.85 bands could be amplified with a primer,averagely.[Conclusion] The selected 20 primers could be effectively applied to ISSR analysis of the germplasm resources of C.chekiangoleosa in Zhejiang.