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    果生炭疽菌转录因子CfHac1的BRLZ结构域生物学功能研究

    Functional analysis of BRLZ motif of the transcription factor CfHac1 in Colletotrichum fructicola

    • 摘要:
        目的  炭疽病是油茶的主要病害,导致巨大的经济损失。果生炭疽菌是油茶炭疽病优势致病菌。对果生炭疽菌的转录因子CfHac1的结构域进行分析,为进一步解析转录因子CfHac1调控病菌致病的分子机制奠定基础,对挖掘油茶炭疽病防控新途径具有重要的理论意义。
        方法  采用点突变技术构建转录因子CfHac1结构域缺失载体,通过PEG介导法将该回补载体转化至CfHAC1基因完全缺失突变体的原生质体中,通过博来霉素抗性和荧光筛选获得结构域缺失回补菌株,进一步研究BRLZ结构域在果生炭疽菌中的生物学功能。
        结果  结果发现转录因子CfHac1含有一个碱性亮氨酸拉链结构域(BRLZ),该结构域包含58个氨基酸残基;与野生型菌株和完全互补菌株相比,BRLZ结构域缺失突变株ΔCfhac1ΔBRLZ生长速率显著降低,分生孢子产量显著减少,不能形成附着胞,对内质网压力胁迫更敏感,丧失对油茶叶的致病力,其表型与ΔCfhac1突变体一致。
        结论  上述结果表明,BRLZ是转录因子CfHac1重要的结构域,对CfHac1在果生炭疽菌中行使正常的生物学功能具有重要的调控作用。

       

      Abstract:
        Objective  Colletotrichum fructicola is a major pathogen causing anthracnose on Camellia oleifera, and leads to substantial losses annually. The domain analysis of transcription factor CfHac1 will help us understanding the CfHac1-regulated pathogenic mechanism of the pathogen and provide new insights to control this disease.
        Method  We constructed the domain deletion vector using point mutation technology and introduced it into the ΔCfhac1 mutant through the PEG-mediated protoplast transformation. The strains were selected by bleomycin and fluorescence, then the function was analyzed in C. fructicola.
        Result  The structure prediction revealed that CfHac1 contained one basic region-leucine zipper motif (BRLZ), which contained 58 amino acid residues. Comparing to the wide type and complemented strains, the mycelial growth rate, conidiation and appressorium formation rate of the Cfhac1ΔBRLZ were significantly reduced, and the Cfhac1ΔBRLZ strain was sensitive to dithiothreitol. The pathogenicity test showed that the Cfhac1ΔBRLZ lost the ability to infect C. oleifera leaves, whose phenotype is consistent with ΔCfhac1 mutant.
        Conclusion  The results show that BRLZ is an important domain of CfHac1 and is essential for normal function of CfHac1 in C. fructicola.

       

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