摘要： 以冬枣DNA为研究对象,利用单因素试验,分析了Mg2+浓度、dNTP浓度、Taq酶含量、引物浓度、模板DNA含量以及退火温度对ISSR-PCR扩增的影响,经优化建立了枣属植物最适ISSR-PCR反应体系,25μL反应液中包含1×PCR Buffer、2.0 mmol/L Mg2+、0.25 mmol/L dNTP、1.5 UTaq酶、1.25μmol/L引物和50 ng模板,最适退火温度为58℃。PCR反应体系的建立为应用ISSR标记技术开展枣种群遗传变异分析和构建遗传图谱奠定基础。

关键词: 枣, ISSR, 体系优化

Abstract: Take the genomic DNA of Ziziphus jujuba Mill. cv. Dongzao as a template, the ISSR??PCR reaction system of Chinese jujube was estabblished after analysised the effect of 6 main factors in PCR react ion using the single factor method.The opt imal ISSR??PCR reaction system was determined as follows: 1 PCR Buffer, 2.0 mmol/L Mg²⁺, 0.25mmol/ L dNTP, 15 U Taq polymerase, 125 ??mol/ L primer and 50 ng template DNA in total 25 L reaction volume. The opimal annealing temperature was 58℃. All the result established a foundation for building cultivar fingerprinting and genetic linkage map in Chinese jujube by using ISSR molecular markers.

Key words: Chinese jujube, ISSR, System optimization

中图分类号: 

• S665.1