Application of tetra primer ARMS-PCR approach for detection of Fusarium graminearum genotypes with resistance to carbendazim
Application of tetra primer ARMS-PCR approach for detection of Fusarium graminearum genotypes with resistance to carbendazim
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应用四引物ARMS-PCR方法检测禾谷镰刀菌多菌灵抗性基因型
DOI：
10.1007/s13313-012-0162-2
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发表时间：
2013-01-01
影响因子：
1.4
通讯作者：
Zhou, Mingguo
中科院分区：
文献类型：
--
作者：
Hou, Yiping;Luo, Qingquan;Zhou, Mingguo
建立了四引物ARMS-PCR（扩增耐突变系统- pcr）检测对多菌灵（甲并咪唑氨基甲酸甲酯类杀菌剂）耐药的禾谷镰刀菌分离株。F. graminearum对多菌灵的抗性是由β(2)-微管蛋白基因（FGSG_06611.3）密码子167、198和200的点突变引起的。根据抗性基因β(2)-微管蛋白基因密码子167、198、200的单核苷酸多态性，分别在每个密码子上设计两对引物进行扩增。根据点突变菌株和野生型菌株在琼脂糖凝胶中的不同模式，成功地进行了SNP基因分型。本研究建立的四引物ARMS-PCR技术是一种简单有效的基因型鉴定技术，在多菌灵耐药管理中具有潜在的应用前景。它将帮助研究人员和种植者选择在田间使用的最佳抗真菌策略。
Tetra-primer ARMS-PCR (amplification refractory mutation system-PCR) was developed to detect isolates of Fusarium graminearum with resistance to carbendazim, a methyl benzimidazole carbamate-group fungicide. Resistance to carbendazim in F. graminearum is caused by point mutations at codon 167, 198, and 200 of the beta (2)-tubulin gene (FGSG_06611.3). According to single nucleotide polymorphisms on codon 167, 198, 200 of the resistance gene beta (2)-tubulin gene, two pairs of primer designed on each codon respectively, were used for amplification. SNP genotyping was made successfully based on the different patterns in agarose gels between point mutations and wild type strain. Tetra-primer ARMS-PCR protocol developed in this study is a simple and useful technique to identify genotypes and has a potential application in management of carbendazim resistance in F. graminearum. It will help researchers and growers select the best antifungal strategy to use in the field.