摘要： 为了获得最优柑橘黑斑病病原菌柑橘叶点霉菌ISSR-PCR反应体系,利用单因素试验法对反应体系中的5个因素(Mg²⁺、dNTP、引物、Taq DNA聚合酶和模板DNA)分别设置了不同的浓度梯度进行反应体系的优化。结果表明,在20μL反应体系中,含有1.5 mmol/L的Mg²⁺、0.15 mmol/L的dNTP、0.3μmol/L的引物、25 ng DNA模板、0.5 U Taq DNA聚合酶为最优。利用优化的ISSR-PCR反应体系对不同地理来源的21株柑橘叶点霉菌菌株进行扩增。结果表明,已建立的体系稳定可靠。柑橘叶点霉菌ISSR-PCR反应体系的建立为利用ISSR分子标记技术对柑橘叶点霉菌进行遗传分析奠定了基础。

关键词: 柑橘叶点霉菌, ISSR, 单因素试验法, 柑橘黑斑病

Abstract: In order to obtain the optimization of ISSR-PCR reaction system of Phyllosticta citricarpa (McAlpine) van der Aa,the concentrations of five factors (Mg²⁺,dNTP,primers, Taq DNA polymerase,template) were respectively optimized by single factor test.The result showed that the optimized reaction system consisted of 1.5 mmol/L Mg²⁺,0.15 mmol/L dNTP,0.3 μmol/L primers,25 ng template DNA,0.5 U Taq DNA polymerase.The optimized ISSR-PCR reaction system was stable and credible by the amplification of 21 strains of P.citricarpa.The study would provide the basis for the genetic analysis of P.citricarpa.

Key words: Phyllosticta citricarpa, Inter-simple sequence repeat, Single factor test, Citrus black spot

中图分类号: 

• S435.79