【摘要】 根据正交实验理论,设计一种优化RAPD各个反应组分以获得理想图谱的最佳反应浓度方法,能够显著减少反应组分优化次数。大丽轮枝菌(Verticilliumdahliae )RAPD扩增最优反应组分浓度为:MgCl2 3.5mmol/L,dNTP0.25mmol/L,引物15pmol/L,Taq酶1.5U,DNA5ng。实验表明,MgCl2与dNTP浓度之比是影响RAPD扩增效果的最主要因素。根据优化的反应组分,用9个随机引物对来自江苏南通、射阳、泗阳、盐都4个群体53个大丽轮枝菌菌株进行RAPD扩增,对各个群体的遗传结构分析表明,各地群体遗传相似性较高。更多还原

【Abstract】 An optimization strategy for RAPD fingerprinting was designed based on orthogonal test, which remarkably reduced the number of experiments. Optimum conditions for RAPD with the maximum number of score products were determined as 3.5 mmol/L MgCl2, 0.25 mmol/L dNTP, 15 pmol/L primer and 5 ng DNA for Verticillium dahliae. The results showed that the proportion of MgCl2 to dNTP concentration was the main factor for RAPD fingerprinting. Under the optimum conditions, nine primers were used to amplify DNA of all 53 isolates of V.dahliae from 4 cotton areas in Jiangsu Province.The analysis indicated that the genetic structures of four populations were similar.更多还原