稻曲病病菌Zn_(2)(Ⅱ)Cys6型转录因子UvZC1基因的克隆及功能分析

Clone and functional research of Zn_(2)(Ⅱ)Cys6 transcription factor UvZC1 gene in Ustilaginoidea virens

为明确Zn_(2)(Ⅱ)Cys6型转录因子UvZC1在稻曲病病菌中的功能,利用CRISPR-Cas9结合同源片段双交换的方法,诱导野生型菌株Jt209发生UvZC1基因缺失突变。结果显示,与Jt209相比,UvZC1基因缺失突变体的生长速率和分生孢子量均显著下降,且突变体中部分与稻曲病病菌其他产分生孢子相关基因表达量发生变化;此外,该基因的缺失导致突变体对十二烷基硫酸钠(SDS)更敏感,而对氧化胁迫的耐受性增强;在稻曲病病菌接种水稻后24 h、48 h的侵染早期,UvZC1基因的表达量明显上升,但基因缺失突变体接种水稻后形成的稻曲球数量与野生型之间没有差异。综上所述,UvZC1基因参与稻曲病病菌营养生长、分生孢子产生和侵染水稻过程,还与稻曲病病菌细胞壁的完整性和响应氧化胁迫相关。

In order to clarify the function of Zn_(2)(Ⅱ)Cys6 transcription factor UvZC1 in Ustilaginoidea virens,gene deletion mutation was carried out by the CRISPR-Cas9-based homologous recombination system.Compared with the wild-type strain Jt209,the growth rate and conidia production of△UvZC1 mutants were significantly decreased,and the expression levels of genes related to the sporulation of U.virens were also changed.In addition,the UvZC1 deletion mutants showed more sensitivity to sodium dodecyl sulfate(SDS),and the tolerance to oxidative stress was enhanced.At the early stage of infection(24 h and 48 h after inoculation),the expression of UvZC1 gene increased significantly.However,no significant difference in the number of rice false smut balls between the wild type and the△UvZC1 mutants was found.Overall,UvZC1 gene has roles in regulating hyphal growth,conidiation,cell wall integrity and oxidative stress response in U.virens.Furthermore,UvZC1 gene is also involved in the infection process of U.virens.