水稻恶苗病菌(Fusarium fujikuroi)β-微管蛋白基因克隆及与多菌灵抗药性关系  

Cloning β-tubulin from Fusarium fujikuroi to analyze its relationship with carbendazim resistance

作　　者：马晓伟[1] 邢春杰[1] 于金凤[1] 王勇[1] 陈子豪[1] 王建新[1] 周明国[1] 陈长军[1] 

机构地区：[1]南京农业大学植物保护学院,江苏省农药学重点实验室,南京210095

出　　处：《微生物学报》2012年第5期581-587,共7页Acta Microbiologica Sinica

基　　金：国家自然科学基金项目(31171880,30971891)~~

摘　　要：【目的】揭示水稻恶苗病菌(Fusarium fujikuroi)对多菌灵的抗药性与其β-微管蛋白基因的相关性。【方法】结合形态学和TEF-1α基因序列对分离菌株进行鉴定;根据近源种拟轮枝镰孢菌(Fusarium verticillioides)核基因组测序菌株7600的β-微管蛋白核苷酸序列设计引物,采用PCR方法克隆并比对分析了F.fujikuroi对多菌灵不同敏感性表型的5个菌株的β-微管蛋白基因全序列;利用实时定量技术(qRT-PCR)分析了β-微管蛋白基因在上述5个菌株中的表达特性。【结果】F.fujikuroi的β-微管蛋白基因核苷酸序列(GenBank登录号:JQ026022)全长1671 bp,包含4个内含子,编码447个氨基酸残基;2个敏感性菌株和3个抗药性菌株的β-微管蛋白基因核苷酸序列同源性100%;在无药剂处理下该基因在2个敏感性菌株中的表达水平显著高于3个抗药性菌株(p=0.05),且对同一菌株而言,药剂处理能够显著提高β-微管蛋白基因表达水平(p=0.05),但在相同药剂处理条件下,菌株间差异不显著。【结论】F.fujikuroi对多菌灵的抗药性机制与β-微管蛋白无关,有待进一步研究。[Objective] We studied the resistance of Fusarium fujikuroi against carbendazim,to confirm whether the resistance was related to the β-tubulin gene.[Methods] The isolated strains were identified based on TEF-1α gene sequence and morphological characteristics.The whole nucleotide sequence of β-tubulin gene was analyzed from different sensitive strains by the primers designed according to the nucleotide sequence of β-tubulin gene from the sequenced strain 7600 of F.verticillioides.Real-time quantitative RT-PCR（qRT-PCR） was applied to analyze the expression pattern of β-tubulin gene from different sensitive strains treated with or without carbendazim.[Results] The full-length nucleotide sequence of β-tubulin gene（Accession Number： JQ026022） was cloned from different sensitive strains of F.fujikoroi,which spanned 1671 bp with 4 introns,encoding 447 amino acids.This gene showed 100% nucleotide sequence homology between sensitive and resistant strains.The expression of the β-tubulin gene in 2 sensitive strains was significantly higher than 3 resistant strains when these strains were cultured on carbendazim-free media（p=0.05）.Carbendazim at the concentration of their EC50 caused the expression of the gene from the five strains increasing significantly（p=0.05） in comparison with that of strains treated without carbendazim.However,there was no significant difference between strains when treated with carbendazim.[Conclusion] The resistance of F.fujikuroi against carbendazim was irrelative to β-tubulin gene.

关 键 词：水稻恶苗病菌 β-微管蛋白基因 表达特性 多菌灵抗药性 

分 类 号：S435.111.44[农业科学—农业昆虫与害虫防治]