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Molecular characterization of Rhizoctonia solani AG4 using PCR-RFLP of the rDNA-ITS region

机译:利用rDNA-ITS区的PCR-RFLP分子分析对茄红假单胞菌AG4的分子表征

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摘要

Rhizoctonia solani isolates obtained from common beans (Phaseolus vulgaris L.) were included in an AG4 anastomosis group in accordance with hyphal anastomosis. In the subgrouping of AG4 isolates, PCR-RFLP patterns in the rDNA-ITS were used. After obtaining the genomic DNA belonging to R. solani AG4, an approximately 700 bp amplification product of the ITS1-5.8S-ITS2 region was obtained with PCR, using ITS1 and ITS4 universal primers. The PCR products were digested with MseI, HincII, AvaII, and MfeI restriction endonucleases, and different PCR-RFLP patterns were obtained for the AG4 HGI and AG4 HGII subgroups. In this study, in addition to the enzymes that were previously used in the AG4 subgrouping, AvaII restriction endonuclease was also seen to be effective in the subgrouping. In this way, the isolates that were grouped as R. solani AG4 according to anastomosis reactions were separated into 2 subgroups, HGI and HGII. This study indicates that the common beans in the Black Sea coastal region are more commonly infected by Rhizoctonia solani AG4 HGI.
机译:根据菌丝吻合,从普通豆(菜豆)获得的茄根枯菌分离株被包括在AG4吻合组中。在AG4分离物的亚组中,使用了rDNA-ITS中的PCR-RFLP模式。在获得了属于茄红假单胞菌AG4的基因组DNA之后,使用ITS1和ITS4通用引物通过PCR获得了ITS1-5.8S-ITS2区域的大约700bp的扩增产物。用MseI,HincII,AvaII和MfeI限制性核酸内切酶消化PCR产物,并为AG4 HGI和AG4 HGII亚组获得不同的PCR-RFLP模式。在这项研究中,除了先前在AG4子组中使用的酶外,还发现AvaII限制性核酸内切酶在该子组中有效。通过这种方式,根据吻合反应将分离株R. solani AG4分为两个亚组:HGI和HGII。这项研究表明,黑海沿岸地区的普通豆类更容易受到茄根枯菌AG4 HGI的感染。

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