舞毒蛾ISSR反应体系的优化与建立  

Optimization and Establishment of ISSR Reaction System for Gypsy Moth(Lymantria dispar Linnaeus)

作　　者：陈芳[1] 骆有庆[1] 石娟[1] 邬颖 杜秋洋 

机构地区：[1]北京林业大学林学院,北京100083

出　　处：《中国农学通报》2014年第10期42-45,共4页Chinese Agricultural Science Bulletin

基　　金：中央高校基本科研业务费专项"生态林重大虫灾监测与防控"(TD2011-06));国家自然科学基金面上项目(31170613);教育部新世纪优秀人才支持计划(NCET-10-0232)

摘　　要：舞毒蛾是一种食性广、危害大的林业害虫,遗传多样性的研究有利于揭示不同地理种群的遗传变异情况。以河北舞毒蛾为实验材料,对ISSR反应体系中的5个因子(Taq DNA聚合酶、模板DNA、引物、MgCl2和dNTPs)的8个浓度梯度依次进行试验,每一因子的最佳浓度都用于下一组优化试验,最终确定最佳反应体系优化筛选,结果表明,20μL ISSR反应体系各组分的最适浓度分别为0.5 U Taq DNA聚合酶,1.5 ng/μL模板DNA,2.4μmol/L引物,1.75 mmol/L MgCl2,0.28 mmol/L dNTPs。这一优化的舞毒蛾ISSR-PCR反应体系为进一步利用ISSR分子标记技术对舞毒蛾的遗传多样性分析奠定了良好的试验基础。The gypsy moth, Lymantria dispar Linnaeus （Lepidoptera： Lymantriidae）, is a destructive defoliator with a broad host range. Studying genetic variation is a helpful tool for revealing the genetic differentiation of different geographic populations of gypsy moth. 5 main factors of ISSR reaction system for gypsy moth （Lymantria dispar Linnaeus） was optimized with sample from Hebei Province, China. Factors with 8 concentration gradients were performed successively. Each optimum concentration of a factor was applied to the next optimization experiment. The final ISSR reaction system for gypsy moth was as follows： 0.5 U Taq DNA polymerase, template DNA 1.5 ng/μL, primer 2.4 μmol/L, 1.75 mmol/L MgCl2. 0.28 mmol/L dNTPs. Then ddH20 was added to the final volume of 20 μL. This optimum reaction system would lay fundamental basis for the genetic diversity and variation research for gypsy moth.

关 键 词：ISSR 舞毒蛾 体系优化 

分 类 号：S718.7[农业科学—林学]