Position: Home > Articles > Optimization of Inter-simple Sequence Repeat PCR Reaction System of Phyllosticta citricarpa
Acta Agriculturae Boreali-Sinica
2016,31
(1)
123-127
柑橘叶点霉菌ISSR-PCR反应体系的优化
作 者：
王兴红;李红叶
单 位：
浙江大学农业与生物技术学院;中国林业科学研究院华北林业实验中心
关键词：
柑橘叶点霉菌;ISSR;单因素试验法;柑橘黑斑病
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+、d NTP、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d NTP、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。
译 名：
Optimization of Inter-simple Sequence Repeat PCR Reaction System of Phyllosticta citricarpa
作 者：
WANG Xinghong;LI Hongye;Experimental Center of Forestry in North China,Chinese Academy of Forestry;College of Agriculture & Biotechnology,Zhejiang University;
关键词：
Phyllosticta citricarpa;;Inter-simple sequence repeat;;Single factor test;;Citrus black spot
摘 要：
In order to obtain the optimization of ISSR-PCR reaction system of Phyllosticta citricarpa( Mc Alpine) van der Aa,the concentrations of five factors( Mg2 +,d NTP,primers,Taq DNA polymerase,template) were respectively optimized by single factor test. The result showed that the optimized reaction system consisted of 1. 5mmol / L Mg2 +,0. 15 mmol/L d NTP,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.
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