舞毒蛾ISSR反应体系的优化与建立

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。

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

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.