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
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国家自然科学基金项目(31171880,30971891)


Cloning β-tubulin from Fusarium fujikuroi to analyze its relationship with carbendazim resistance
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Supported by the National Natural Science Foundation of China(31171880,30971891)

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    摘要:

    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1671bp,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。

    Abstract:

    Abstract:[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.

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马晓伟,邢春杰,于金凤,王勇,陈子豪,王建新,周明国,陈长军. 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. 微生物学报, 2012, 52(5): 581-587

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  • 收稿日期:2011-12-08
  • 最后修改日期:2012-02-26
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  • 在线发布日期: 2012-05-11
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