CAJ | 학술논문

为建立一种简单、快捷的方法以鉴定梨种质资源，对CTAB法、SDS法和高盐法3种DNA提取方法进行了比较．结果表明，改良的CTAB法较为适合梨基因组DNA的提取．通过对TaqE，Mg^＋，dNTP“模板DNA”和primer不同浓度的优化实验以及对PCR反应程序的优化筛选，建立了适合梨基因组ISSR分析的技术体系．运用该体系对梨品种进行ISSR分析，结果表明，从100对引物中筛选出的10对引物能扩增清晰带且多态性好，从而达到鉴定梨种质资源的目的，并且应用该体系构建了新疆主要梨的遗传图谱和数字指纹图谱．
위건립일충간단、쾌첩적방법이감정리충질자원，대CTAB법、SDS법화고염법3충DNA제취방법진행료비교．결과표명，개량적CTAB법교위괄합리기인조DNA적제취．통과대TaqE，Mg^＋，dNTP“모판DNA”화primer불동농도적우화실험이급대PCR반응정서적우화사선，건립료괄합리기인조ISSR분석적기술체계．운용해체계대리품충진행ISSR분석，결과표명，종100대인물중사선출적10대인물능확증청석대차다태성호，종이체도감정리충질자원적목적，병차응용해체계구건료신강주요리적유전도보화수자지문도보．
Three DNA extraction methods, CTAB, SDS and high salt method, were compared in this experiment to find a simple and rapid one to identify the pear germplasm. The results showed that the improved CTAB method was the best one to extract pear gonomic DNA. Well-purified DNA was obtained and qualified for ISSR-PCR analysis. Different PCR program and the ingredients in ISSR reaction system, such as TaqE, Mg＋, dNTP, DNA template and primer, were experimentally selected by different concentrations and suitable reaction system. And program of the ISSR analysis for pear genomics was settled. DNA fingerprints were also obtained in all of the tested species, which indicated that the ISSR technology could be applied in the studies of pear germplasm in genetic mapping, identification of fingerprints, molecular marker and so on.