大丽轮枝菌侵染陆地棉早期的转录组分析

doi:10.11963/1002-7807.zzdzkj.20170508

摘要/Abstract

摘要： 目的旨在深入挖掘大丽轮枝菌致病相关基因。方法运用转录组测序技术比较分析了强致病力大丽轮枝菌Vd991在侵染陆地棉品种Coker 312早期阶段的基因表达情况。结果对Vd991侵染前后的转录组数据进行差异表达分析,共筛选到979个差异表达基因（Differentially expressed genes,DEGs）,其中376个上调,603个下调。通过基因注释和功能分类发现,这些DEGs涉及细胞增殖与生长、产孢、碳水化合物结合、蛋白激酶活性调节、裂解酶活性、水解酶活性等功能。对上调DEGs进行基因本体（Gene ontology,GO）功能富集分析,发现共有277个GO词条达到显著富集水平(P<0.05),涉及生长速率正向调控、核苷酸合成、解旋酶活性等功能;京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes,KEGG）通路分析表明,上调DEGs参与53个代谢通路,包括嘧啶代谢、嘌呤代谢等。结论在早期侵染过程中,大丽轮枝菌细胞增殖相关基因表现活跃。上述分析为进一步揭示大丽轮枝菌的致病机理提供了有价值的参考。

关键词: 大丽轮枝菌; 转录组分析; 细胞增殖; 致病机理

Abstract:

[Objective] The aim of this study is to mine pathogenesis related genes of Verticillium dahliae. [Method] We explored the transcriptome of highly toxic V. dahliae strain Vd991 at the early stage of infection in Coker 312, an upland cotton cultivar. [Result] Through differential expression analysis, a total of 979 differentially expressed genes (DEGs）were identified after infection, of which 376 were up-regulated and 603 were down-regulated. The result of gene annotation and classification showed that these DEGs are involved in broad biological processes, molecular function and cellular component, including reproductive process, growth, sporulation, carbohydrate binding, kinase regulator activity, lyase activity and hydrolase activity. Enrichment analysis of the up-regulated DEGs were performed, 277 gene ontology terms were enriched, including positive regulation of growth rate, purine nucleotide biosynthetic process, pyrimidine nucleobase biosynthetic process, helicase activity. A total of 53 Kyoto Encyclopedia of Genes and Genomes pathways were identified in the up-regulated DEGs, including purine metabolism, pyrimidine metabolism. [Conclusion] These results indicate that genes related to cell proliferation actively expressed at the early stage of infection and provide valuable reference for exploring the pathogenic mechanism of V. dahliae.

Key words: Verticillium dahliae; transcriptome analysis; cell proliferation; pathogenic mechanism

张志东,段兴鹏,周玉梅,李建福,韩松,高正银,赵佳薇,左开井. 大丽轮枝菌侵染陆地棉早期的转录组分析[J]. 棉花学报, 2017, 29(3): 253-260.

Zhidong Zhang,Xingpeng Duan,ZhouYumei,Jianfu Li,Song Han,Zhengyin Gao,Jiawei Zhao,Kaijing Zuo. Transcriptome Analysis of Verticillium dahliae in Upland Cotton at Early Infection Stage by RNA-seq[J]. Cotton Science, 2017, 29(3): 253-260.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: http://journal.cricaas.com.cn/Jweb_mhxb/CN/10.11963/1002-7807.zzdzkj.20170508

http://journal.cricaas.com.cn/Jweb_mhxb/CN/Y2017/V29/I3/253

参考文献

[1]	徐理, 朱龙付, 张献龙. 棉花抗黄萎病机制研究进展[J]. 作物学报, 2012, 3(9): 1553-1560.
[1]	Xu Li, Zhu Longfu, Zhang Xianlong.Research on resistance mechanism of cotton to Verticillium wilt[J]. Acta Agronomica Sinica, 2012, 3(9): 1553-1560.
[2]	简桂良, 邹亚飞, 马存. 棉花黄萎病连年流行的原因及对策[J]. 中国棉花, 2003, 30(3): 13-14.
[2]	Jian Guiliang, Zou Yafei, Ma Cun.The reason of the prevalence of Verticillium wilt of cotton in consecutive years and the corresponding countermeasures[J]. China Cotton, 2003, 30(3): 13-14.
[3]	朱荷琴, 冯自力, 刘雪英, 等. 转基因棉田黄萎病发生特点及控制技术[J]. 中国棉花, 2009, 36(5): 20-21.
[3]	Zhu Heqin, Feng Zili, Liu Xueying, et al.Occurrence and control technology of transgenic cotton Verticillium wilt[J]. China Cotton, 2009, 36(5): 20-21.
[4]	Rauyaree P, Ospina G M D, Kang S, et al. Mutations in VMK1, a mitogen-activated protein kinase gene, affect microsclerotia formation and pathogenicity in Verticillium dahliae[J]. Current Genetics, 2005, 48(2): 109-116.
[5]	Klimes A, Dobinson K F.A hydrophobin gene, VDH1, is involved in microsclerotial development and spore viability in the plant pathogen Verticillium dahliae[J]. Fungal Genetics and Biology, 2006, 43(4): 283-294.
[6]	Tzima A K, Paplomatas E J, Rauyaree P, et al.Roles of the catalytic subunit of cAMP-dependent protein kinase A in virulence and development of the soilborne plant pathogen Verticillium dahliae[J]. Fungal Genetics and Biology, 2010, 47(5): 406-415.
[7]	Tzima A K, Paplomatas E J, Tsitsigiannis D I, et al.The G protein β subunit controls virulence and multiple growth- and development- related traits in Verticillium dahliae[J]. Fungal Genetics and Biology, 2012, 49(4): 271-283.
[8]	Zhou Bangjun, Jia Peisong, Gao Feng, et al.Molecular characterization and functional analysis of a necrosis- and ethylene-inducing, protein-encoding gene family from Verticillium dahliae[J]. Molecular Plant-Microbe Interactions, 2012, 25(7): 964-975.
[9]	Tzima A K, Paplomatas E J, Rauyaree P, et al.VdSNF1, the sucrose nonfermenting protein kinase gene of Verticillium dahliae, is required for virulence and expression of genes involved in cell wall degradation[J]. Molecular Plant-Microbe Interactions, 2011, 24(1): 129-142.
[10]	田李. 大丽轮枝菌基因敲除体系的建立及分泌蛋白的初步分析[D]. 北京: 中国农业科学院, 2011.
[10]	Tian Li.Construction of high efficient gene knockout system and functional analysis of secreted protein in Verticillium dahliae[D]. Beijing: Chinese Academy of Agricultural Sciences, 2011.
[11]	Liu Shaoyan, Chen Jieyin, Wang Jinlong, et al.Molecular characterization and functional analysis of a specific secreted protein from highly virulent defoliating Verticillium dahliae[J]. Gene, 2013, 529: 307-316.
[12]	Santhanam P, Thomma B P H J. Verticillium dahliae Sge1 differentially regulates expression of candidate effector genes[J]. Molecular Plant-Microbe Interactions, 2013, 26(2): 249-256.
[13]	Trapnell C, Williams B A, Pertea G, et al.Transcript assembly and quantification by RNA Seq reveals unannotated transcripts and isoform switching during cell differentiation[J]. Nature Biotechnology, 2010, 28(5): 511-515.
[14]	Mortazavi A, Williams B A, McCue K, et al. Mapping and quantifying mammalian transcriptomes by RNA-Seq[J]. Nature Methods, 2008, 5(7): 621-628.
[15]	Anders S, Huber W. Differential expression analysis for sequence count data[J/OL]. Genome Biology, 2010, 11(10): R16 [2016-09-28]. . DOI: 10.1186/gb-2010-11-10-r106.
[16]	Ye Jia, Fang Lin, Zheng Hongkun, et al.WEGO: a web tool for plotting GO annotations[J]. Nucleic Acids Research, 2006, 34: 293-297.
[17]	肖红利. 棉花组织诱导体系中大丽轮枝菌分泌蛋白分析及其致病性研究[D]. 北京: 中国农业科学院, 2014.
[17]	Xiao Hongli.Quantitative secretome analysis and pathogenicity study of Verticillium dahliae in cotton-extract induction system[D]. Beijing: Chinese Academy of Agricultural Sciences, 2014.
[18]	Klosterman S J, Subbarao K V, Kang S, et al. Comparative genomics yields insights into niche adaptation of plant vascular wilt pathogens[J/OL]. PLoS Pathogens, 2011, 7(7): e1002137 (2011-07-28) [2016-09-28]. .
[19]	W?sten H A B: Hydrophobins: multipurpose proteins[J]. Annual Review of Microbiology, 2001, 55(1): 625-646.
[20]	Kershaw M J, Talbot N J.Hydrophobins and repellents: proteins with fundamental roles in fungal morphogenesis[J]. Fungal Genetics and Biology, 1998, 23(1): 18-33.
[21]	Black J D.Protein kinase C-mediated regulation of the cell cycle[J]. Frontiers in Bioscience, 2000, 5: D406-D423.
[22]	Pain V M.Initiation of protein synthesis in eukaryotic in cells[J]. European Journal of Biochemistry, 1996, 236(3): 747-771.
[23]	Dong Zizheng, Zhang Jianting.Initiation factor eIF3 and regulation of mRNA translation, cell growth, and cancer[J]. Critical Reviews in Oncology/Hematology, 2006, 59(3): 169-180.
[24]	Shah M, Su D, Scheliga J S, et al.A transcript-specific eIF3 complex mediates global translational control of energy metabolism[J]. Cell Reports, 2016, 16(7): 1891-1902.
[25]	Lee A S Y, Kranzusch P J, Cate J H D. eIF3 targets cell-proliferation messenger RNAs for translational activation or repression[J]. Nature, 2015, 522(7554): 111-114.