摘要
十字花科蔬菜黑斑病主要是由芸薹生链格孢(Alternaria brassicicola)引起的,通常使蔬菜产质量下降,造成严重损失。Zn_(2)Cys_(6)型锌指蛋白是真菌中特有的转录因子,参与调控真菌生长发育及胁迫应答等。本试验以芸薹生链格孢中编码Zn_(2)Cys_(6)锌指蛋白的转录因子Ab10279基因为研究对象,通过生物信息学分析发现,该基因全长1 069 bp,cDNA全长960 bp,编码319个氨基酸,含有一个GAL4锌指结构。采用基因敲除技术研究Ab10279基因的功能,结果显示Ab10279基因的缺失导致突变体在PDA培养基上生长速度高于野生菌,在MM培养基上低于野生菌,且突变体菌落为米白色;显微镜下观察,Ab10279基因缺失突变体在PDA培养基上未见分生孢子,链格出现膨大现象,在MM培养基上分生孢子链格数量增多;缺失突变体的分生孢子在24 h后萌发率降低,芽管长度和芽管分叉数增多;突变体在寄主小油菜叶片上致病性下降,病斑面积较野生菌降低40%左右,但突变体菌丝在穿透小油菜叶片后可继续生长。将Ab10279基因回补后,互补株C与野生菌表现几乎一致。
Alternaria brassicicola is the main pathogenic fungus causing black spot of cruciferous vegetable.Black spot usually causes the decline of yield and quality of cruciferous vegetable,causing serious losses.Zn_(2)Cys_(6)zinc finger protein is a unique transcription factor in fungi,which is involved in the regulation of fungal;growth and development and stress response.,The research object was the transcription factor Ab10279 gene encoding Zn_(2)Cys_(6)zinc finger protein in A.brassicicola in this study.Bioinformatics analysis showed that the full length of the gene was 1 069 bp and the cDNA is 960 bp,encoding 319 amino acids containing a zinc finger structure of GAL4.In this study,gene knockout technology was used to study the function of Ab10279 gene.The results showed that the Ab10279 gene deletion mutant grew faster than wild bacteria on PDA medium and lower than wild bacteria on MM medium,and the colony morphology of mutant was off-white.Under microscopic observation,no conidia were observed in Ab10279 gene deletion mutant on the PDA medium,the chain grids were enlarged,and the number of conidia chain grids increased on the MM medium.The conidia germination rate of the mutant decreased after 24 h,and the length and bifurcation number of germ tube increased.The pathogenicity of Ab10279 gene deletion mutants was decreased,and the lesion area was about 40%lower than that of wild type.The mutants could continue to grow after penetrating the leaves of Brassica napes.The performance of the complementary strain C was almost the same as that of the WT after the Ab10279 gene was complemented.
作者
毛胜洁
张敏
李珊珊
刘琛
徐后娟
Mao Shengjie;Zhang Min;Li Shanshan;Liu Chen;Xu Houjuan(College of Botany,Shandong Agricultural University,Tai'an,271018)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第11期3548-3557,共10页
Genomics and Applied Biology
基金
山东省现代农业产业技术体系烟草创新团队项目(SDAIT-25-07)资助
关键词
芸薹生链格孢
ZN
CYS
转录因子
产孢
致病性
Alternaria brassicicola
Zn2Cys6 transcription factor
Conidiation
Pathogenicity
