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为了解38株尖镰孢菌的遗传多样性并开发可在近缘镰孢菌种中通用的尖镰孢菌EST-SSR标记,利用设计和筛选的18对多态性EST-SSR引物对38株尖镰孢菌和5种近缘镰孢菌进行SSRPCR扩增,经6%非变性聚丙烯酰胺凝胶分离扩增产物,并用NTSYS软件分析供试尖镰孢菌的PCR扩增结果。结果表明,18对EST-SSR标记引物在38株尖镰孢菌中检测到75条多态性条带,多态性比率达92.6%,平均每对引物可扩增4.2条;各菌株间的遗传相似系数介于0.565~0.946之间,平均为0.721;来源于同科寄主植物群体的菌株间的平均遗传相似系数以葫芦科最大,锦葵科最小,依次为葫芦科>兰科>豆科>亚麻科>茄科>锦葵科。在相似系数为0.756时,供试38株菌有35株按照不同科寄主植物聚为不同的类群,说明尖镰孢菌SSR类群的划分与其寄主来源具有一定的相关性。18对EST-SSR引物在近缘镰孢菌种中均能有效扩增的引物数及通用性比率为10对和55.6%,均显示多态性的引物有2对,占供试引物总数的11.1%。表明尖镰孢菌ESTSSR区域遗传多样性丰富,基于尖镰孢菌EST序列开发镰孢菌通用SSR标记是可行的。
To understand the genetic diversity of 38 Fusarium culms and to develop Fusarium oxysporum EST-SSR markers that can be used universally in Fusarium species, 18 pairs of polymorphic EST-SSR primer pairs designed and screened 38 Fusarium spp. And five species of Fusarium spp. Were amplified by SSRPCR. The amplification products were separated on a 6% non-denaturing polyacrylamide gel and analyzed by PCR with NTSYS software. The results showed that 18 polymorphic bands were detected in 18 Fusarium oxysporum with 18 pairs of EST-SSR primers, with a polymorphism rate of 92.6% and an average of 4.2 primers per primer pair. Among the isolates The genetic similarity coefficient ranged from 0.565 to 0.946, with an average of 0.721. The average genetic similarity coefficient of the isolates from the host families of the same families was the largest in Cucurbitaceae and the lowest in Malvaceae, followed by Cucurbitaceae, Orchidaceae, > Flax> Solanaceae> Malvaceae. When the similarity coefficient was 0.756, 35 strains of 38 strains tested were clustered into different groups according to different host plants, indicating that the classification of Fusarium oxysporum SSR was related to their host sources. 18 pairs of EST-SSR primers in the Fusarium species can be effectively amplified in the primer number and the universal ratio of 10 pairs and 55.6%, both showed polymorphic primer pairs, accounting for the total number of primers 11.1%. The results showed that the ESTSSR region of Fusarium oxysporum is rich in genetic diversity. Based on the Fusarium oxysporum EST sequence, it is feasible to develop a common SSR marker of Fusarium.