苹果树腐烂病菌Zn(Ⅱ)2Cys6类转录因子Vmzctf-07的鉴定及其功能分析

Identification and Functional Analysis of Vmzctf-07,A Zn(Ⅱ)_(2)Cys_(6) Transcription Factor from Valsa mali

转录因子是真菌生命过程中重要的调控因子,探究转录因子在苹果树腐烂病菌Valsa mali致病过程中的作用,能够为解析病菌致病机理奠定基础。通过构建Vmzctf-07基因的酵母单杂交和亚细胞定位表达载体,进行转录激活域验证和亚细胞定位分析;通过创制Vmzctf-07基因缺失突变体和回补菌株,进行生长速率、非生物胁迫和致病力测定。结果表明,Vmzctf-07具有转录激活功能,转录激活域可能存在于down区段(aa 1455~aa 1644)。Vmzctf-07定位于细胞核中,符合转录因子典型特征。Vmzctf-07敲除突变体的生长速率和致病力均显著下降,同时该基因参与病菌响应渗透胁迫和氧化胁迫。综上,Vmzctf-07具有转录激活功能和核定位信号,且参与调节V.mali的生长发育、非生物胁迫和致病过程。

Transcription factors(TFs)are important regulators in the life process of fungi.Exploring the role of TFs in the pathogenic process of Valsa mali can lay the foundation for the analysis of pathogenic mechanisms of pathogens.The yeast one-hybrid and subcellular localization expression vectors of Vmzctf-07 were constructed to verify the transcriptional activation domain and analyze the subcellular localization,respectively.The Vmzctf-07 knockout mutants and complement strains were constructed to test the growth rate,response to abiotic stresses and pathogenicity.The results showed that the transcriptional activation domain of Vmzctf-07 may exist in the down segment(aa 1455-aa 1644).Vmzctf-07 was located in nuclear,which accorded with TFs'characteristics.The growth rate and pathogenicity of the knockout mutants were significantly reduced.Meanwhile,Vmzctf-07 was found to be involved in the response to osmotic stress and oxidative stress of V.mali.In summary,Vmzctf-07 has transcriptional activation function and is located in nuclear,and it is involved in regulating the growth,response to abiotic stress and pathogenic process of V.mali.