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为研究烟草野火病菌的群体遗传多样性,采用正交试验方法,对5个多态性引物(REP,ERIC,BOX,J3,IS1112)的Rep-PCR反应体系分别进行了优化,同时优化了体系中的Mg2+、dNTP、rTaq酶等几个因素,并在此基础上对来源于吉林省不同地区的70株烟草野火病菌进行了分子多态性分析。结果表明:优化的PCR反应体系电泳图谱清晰,多态性丰富,重复性好,适合烟草野火病菌群体分子标记研究。通过聚类分析发现,吉林省烟草野火病菌群体存在分子多态性,在遗传相似系数为0.9时,70个菌株被聚为4个类群(G1~G4),其中G1类群最大(39个菌株,占55.71%),而G3类群最小,仅包括2个菌株。G1和G2类群分别包含了5个和3个亚群。不同类群和亚群的菌株地理来源存在多样性。
In order to study population genetic diversity of tobacco wildfire, orthogonal design was used to optimize the Rep-PCR reaction system of five polymorphic primers (REP, ERIC, BOX, J3, IS1112) Mg2 +, dNTPs and rTaq enzymes, and on this basis, 70 tobacco wildfire pathogens from different areas of Jilin Province were analyzed for molecular polymorphism. The results showed that: the optimized PCR reaction system showed clear electrophoresis pattern, rich polymorphism, good repeatability and suitable for the study of molecular markers of tobacco wildfire. Cluster analysis showed that there were molecular polymorphisms in the tobacco wildfire pathogen population in Jilin Province. When the genetic similarity coefficient was 0.9, 70 strains were clustered into four groups (G1 ~ G4), of which the G1 group was the largest (39 strains, Accounting for 55.71%), while the G3 group is the smallest, including only two strains. Groups G1 and G2 contain 5 and 3 subpopulations, respectively. The geographical origin of strains of different taxa and subgroups is diverse.