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【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】 The objective of this study was to reveal the relationship between resistance to carbendazim and its β-tubulin gene in Fusarium fujikuroi. 【Method】 The isolates were identified by combining morphological and TEF-1α gene sequences. Based on the β-tubulin nucleotide sequence of the near-origin Fusarium verticillioides nuclear sequencing strain 7600, primers were designed, The full-length β-tubulin gene of five strains with different sensitive phenotypes of F. fujikuroi was cloned by PCR and analyzed by qRT-PCR Protein gene expression in the above five strains. 【Result】 The nucleotide sequence of the β-tubulin gene of F. fujikuroi (GenBank accession number: JQ026022) was 1671 bp in length and contained 4 introns, which encoded 447 amino acid residues. Two sensitive strains, 3 The homology of nucleotide sequence of β-tubulin gene of 100 resistant strains was 100%. The expression level of β-tubulin gene in 2 sensitive strains was significantly higher than that of 3 resistant strains (p = 0.05 ), And for the same strain, the drug treatment can significantly increase the expression of β-tubulin gene (p = 0.05), but under the same drug treatment conditions, there was no significant difference between strains. 【Conclusion】 The drug resistance mechanism of F.fujikuroi to carbendazim has nothing to do with β-tubulin, which needs further study.