| 212 | 3 | 45 |
| 下载次数 | 被引频次 | 阅读次数 |
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_2Cys6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_2Cys6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。
Abstract：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_2Cys_6zinc finger protein is a unique transcription factor in fungi,which is involved in the regulation of fungal~((1))growth and development and stress response.,The research object was the transcription factor Ab10279 gene encoding Zn_2Cys_6zinc 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.
Brown D.W.,Butchko R.A.,Busman M.,and Proctor R.H.,2007,The Fusarium verticillioides FUM gene cluster encodes a Zn(II)2Cys6 protein that affects FUM gene expression and fumonisin production,Eukaryot Cell,6(7):1210-1218.
Cao H.J.,Zhang J.J.,Du Y.,Qi Z.Q.,Yu M.N.,and Liu Y.F.,2019,Research progress on transcription factors of Magnaporthe grisea,Jiangsu Nongye Xuebao(Jiangsu Journal of Agricultural Sciences),2019,35(06):1493-1500.(曹慧娟,张瑾瑾,杜艳,齐中强,俞咪娜,刘永峰,2019,稻瘟病菌转录因子研究进展,江苏农业学报,2019,35(06):1493-1500.)
Cho Y.,Ohm R.A.,Grigoriev I.V.,and Srivastava A.,2013,Fungal-specific transcription factor AbPf2 activates pathogenicity in Alternaria brassicicola,Plant Journal,75(3):498-514.
Chung H.,Choi J.,Park S.Y.,Jeon J.,and Lee Y.H.,2013,Two conidiation-related Zn(II) 2 Cys 6 transcription factor genes in the rice blast fungus,Fungal Genet.Biol.,61(Complete):133-141.
Hou Z.H.,2020,Expression analysis of the Zn2Cys6transcription factor family of Pleurotus ostreatus and the functional study of PoZCP26,Thesis for M.S.,Graduate School of Institute of Agricultural Resources and Agricultural Regional Planning,Chinese Academy of Agricultural Sciences,Supervisor:Huang C.Y.,pp.44-47.(侯志浩,2020,糙皮侧耳Zn2Cys6转录因子家族的表达分析及PoZCP26的功能研究,硕士学位论文,中国农业科学院农业资源与农业区划研究所研究生院,导师:黄晨阳,pp.44-47.)
Huang J.,Zhang H.S.,and Wang J.F.,2004,The structure and function of plant C2H2 zinc finger protein,Yichuan(Heredity),(03):414-418.(黄冀,张红生,王建飞,2004,植物C2H2型锌指蛋白的结构与功能,遗传,(03):414-418.)
Huang S.S.,Keyhani N.O.,Zhao X.,and Zhang Y.J.,2019,The Thm1 Zn(II)2Cys6 transcription factor contributes to heat,membrane integrity and virulence in the insect pathogenic fungus Beauveria bassiana.Environmental Microbiology,21(8):3153-3171.
Imazaki I.,Kurahashi M.,Iida Y.,and Tsuge T.,2007,Fow2,a Zn(II)2Cys6-type transcription regulator,controls plant infection of the vascular wilt fungus Fusarium oxysporum,Molecular Microbiol,63(3):737-53.
Kang Z.T.,Jiang L.M.,Luo Y.Y.,Liu C.J.,and Li X.R.,2013,The research advances of mechanism of pathogenicity of Alternaria phytopathogenic fungi,Shengming Kexue(Chinese Bulletin of Life Sciences),25(09):908-914.(康子腾,姜黎明,罗义勇,柳陈坚,李晓然,2013,植物病原链格孢属真菌的致病机制研究进展,生命科学,25(09):908-914.)
Li X.,Wang F.T.,Feng J.,Pang Y.X.,Jia X.,Wu Y.Q.,Lang X.W.,Lin R.M.,and Xu S.C.,2019,The role of two zinc finger transcription factor genes in regulating the heat tolerance of wheat stripe rust,Chinese Society for Plant Protection,2019 Academic Annual Meeting of China Society of Plant Protection:18.(李雪,王凤涛,冯晶,庞云星,贾湘,吴艳琴,郎晓威,蔺瑞明,徐世昌,2019,两个锌指转录因子基因在调控小麦条锈菌耐高温反应中的作用,中国植物保护学会,中国植物保护学会2019年学术年会:18.)
Liang T.T.,Wang Y.Y.,Cheng X.J.,Zeng B.,and He B.,2019,Research progress on the structure and function of Zn(II) 2Cys6 zinc finger transcription factor,Jiangxi Kejishifan Daxue Xuebao(Journal of Jiangxi Science and Technology Normal University),(06):96-98.(梁甜甜,王亦婧,程晓婕,曾斌,贺斌,2019,Zn(Ⅱ)2Cys6锌指转录因子的结构和功能研究进展,江西科技师范大学学报,(06):96-98.)
Luan H.D.,Zhan Y.Y.,Jiang Y.Y.,Xu G.B.,and Li Y.R.,2014,Comparison of extraction methods of total RNA from mycelium from chicken-claw-shaped polyporus umbellate of Changbai Mountain,Yanbian Daxue Nongxue Xuebao(Journal of Agricultural Science Yanbian University),36(4):297-301.(栾换东,战莹莹,姜颖越,许广波,李艳茹,2014,长白山药用真菌鸡爪苓菌丝体总RNA提取方法的比较,延边大学农学学报,36(4):297-301.)
Porquier A.,Moraga J.,Morgant G.,Dalmais B.,Simon A.,Sghyer H.,Collado I.G.,and Viaud M.,2019,Botcinic acid biosynthesis in Botrytis cinerea relies on a subtelomeric gene cluster surrounded by relics of transposons and is regulated by the Zn2Cys6transcription factor BcBoa13,Current Genetics,65(4):965-980.
Qiu L.,Zhang J.,Song J.Z.,Hu S.J.,Zhang T.S.,Li Z.,Wang J.J.,and Cheng W.,2021,Involvement of BbTpc1,an important Zn(II)2Cys6 transcriptional regulator,in chitin biosynthesis,fungal development and virulence of an insect mycopathogen,International Journal of Biological Macromolecules,166:1162-1172
Sahu U.,Kris2h na Rao K.,and Rangarajan P.N.,2014,Trm1p,a Zn(II)Cys6-type transcription factor,is essential for the transcriptional activation of genes of methanol utilization pathway,in Pichia pastoris,Biochem.Biophys.Res.Commun.,451(1):158-164.
Tsuji G.,Kenmochi Y.,Takano Y.,Sweigard J.,Farrall L.,Furusawa I.,Horino O.,and Kubo Y.,2000,Novel fungal transcriptional activators,Cmr1p of Colletotrichum lagenarium and pig1p of Magnaporthe grisea,contain Cys2His2 zinc finger and Zn(II)2Cys6 binuclear cluster DNA-binding motifs and regulate transcription of melanin biosynthesis genes in a developmentally specific manner,Mol.Microbiol.,38(5):940-954.
Xiang J.H.,Li L.Z.,and Chen X.B.,2012,Progress in this study of abiotic stress-related zinc finger protein genes in plants,Henong Xuebao(Journal of Nuclear Agricultural Sciences),26(04):666-672,716.(向建华,李灵之,陈信波,2012,植物非生物逆境相关锌指蛋白基因的研究进展,核农学报,26(04):666-672,716.)
Xu H.,Wang Y.,Zhao P.,Zhang Y.and Li D.,2011,AcAMP-Dependent protein kinase gene,Aapk1,is required for mycelia growth,toxicity and pathogenicity of Alternaria alternata on tobacco,J.Phytopathol.159(4):208-216.
Yusof M.T.B.,Kershaw M.J.,Soanes D.M.,and Talbot N.J.,2014,FAR1 and FAR2 regulate the expression of genes associated with lipid metabolism in the rice blast fungus Magnaporthe oryzae,PLoS ONE,9(6):e99760.
Zhang X.X.,Wang L.,and Shou L.L.,2012,An improved CTAB method for rapid extraction of wheat genomic DNA,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),28(36):46-49.(张晓祥,王玲,寿路路,2012,一种快速提取小麦基因组DNA的改良CTAB方法,中国农学通报,28(36):46-49.)
基本信息:
DOI：10.13417/j.gab.040.003548
中图分类号:S432.44
引用信息:
[1]毛胜洁,张敏,李珊珊,等.芸薹生链格孢(Alternaria brassicicola)Ab10279基因的功能分析[J].基因组学与应用生物学,2021,40(Z4):3548-3557.DOI:10.13417/j.gab.040.003548.
基金信息:
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
2021-12-25
2021-12-25