摘要： 为了对玉米弯孢病菌的鉴定、区域发生关系以及遗传多样性等进行研究，采用单因素试验优化玉米弯孢叶斑病菌ISSR-PCR反应体系中模板DNA、引物、dNTPs、Taq酶、Mg²⁺的用量，并确定每条引物的最佳退火温度。结果表明，最佳反应体系为：模板DNA 1.6 μL（25 ng/μL）、引物1.4 μL（5 μmol/L）、dNTPs 0.25 μL（2.5 mmol/L）、Taq酶0.4 μL（2.5 U/μL）、Mg²⁺ 0.8 μL（25 mmol/L）、10×Taq Buffer 2 μL、ddH₂O 13.55 μL。在该体系下选用3个玉米弯孢病菌的全基因组DNA，分别对52条ISSR引物进行筛选，筛选出16条多态性高、扩增稳定的ISSR引物。玉米弯孢病菌ISSR-PCR反应体系的建立为利用ISSR分子标记技术对该病原菌进行遗传分析奠定了基础。

关键词: 玉米弯孢病菌, ISSR-PCR, 体系优化

Abstract: To study the identification,regional occurrence correlation and genetic diversity of Maize curvularia pathogens,single factor test was employed to optimize the quantities of the template DNA,primer,dNTPs,Taq polymerase and Mg²⁺ used in ISSR-PCR,as well as the determination of the optimum annealing temperature of each primer.The optimal reaction system was established in which template DNA,primer,dNTPs, Taq polymerase,Mg²⁺×10 Taq Buffer and ddH₂O were 1.6 μL(25 ng/μL),1.4 μL (5 μmol/L),0.25 μL (2.5 mmol/L),0.4 μL (2.5 U/μL),0.8 μL (25 mmol/L),2 μL,13.55 μL respectively.52 primers for ISSR were screened with optimized system using genomic DNA of 3 Curvularia strains, and 16 primers with stable amplification and rich polymorphism were obtained.The study would provide the basis for the genetic analysis of maize Curvularia pathogens.

Key words: Maize curvularia pathogens, ISSR-PCR, Optimization reaction system

中图分类号: 

• S435.13