球孢白僵菌基因组候选效应子的预测与分析
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摘要
球孢白僵菌(Beauveria bassiana)是一种广泛应用的生防菌株,效应子在其对寄主昆虫侵染过程中发挥着重要作用。为高通量地筛选球孢白僵菌的效应子,本研究根据球孢白僵菌全基因组信息,利用SignalP、TMHMM、TargetP和Protcomp等生物信息学软件和预测程序对球孢白僵菌中10 364个蛋白序列进行分泌蛋白预测。结果表明,共分析到940个分泌蛋白,包含了185个碳水化合物活性酶家族蛋白。其中碳水化合物结合模块(CBM) 40个,碳水化合物酯酶(CE) 23个,糖苷水解酶(GH) 92个,糖基转移酶(GT) 4个,多糖裂解酶(PL) 2个,辅助功能酶(AA) 24个。进一步将上述分泌蛋白与胞外酶数据库进行比对分析、半胱氨酸含量分析、多个串联重复序列分析、病原寄主互作数据库进行比对及剔除大于300个氨基酸序列的蛋白,共获得18个候选效应子,其中11个为功能未知的假定蛋白,其余为胞外蛋白、细胞壁蛋白、几丁质酶蛋白等。本研究结果为进一步探究效应子在病原菌与昆虫寄主中的互作奠定了理论基础,并为其他病原菌效应子的预测及分析提供了技术参考。
Beauveria bassiana is wildly used in biological control. Effectors played an important role in the infection process. SignalP、TMHMM、TargetP and Protcomp bioinformatics methods were used to analysis the B. bassiana genome that contain 10 364 protein,totally 940 secreted proteins were obtained,including 185 carbohydrate active enzyme proteins. Among them,there are 40 carbohydrate binding modules,23 carbohydrate esterases,92 glycoside hydrolases,4 glycosyltransferases,2 polysaccharides lysases,and 24 auxiliary activities. Furtherly the above secretory proteins were blast with the extracellular enzyme database the cysteine content and multiple tandem repeats were analyzed. Compared with the Pathogen-Host interaction database ant then screened out the proteins larger than 300 amino acid. Totally 18 proteins were confirmed to be the effectors of B. bassiana. This study used bioinformatics to predict candidate effectors from B. bassiana genomic,laid a foundation for further research on interaction between pathogen and insect host,and provide reference for the prediction and analysis of other pathogen effectors.
Beauveria bassiana is wildly used in biological control. Effectors played an important role in the infection process. SignalP、TMHMM、TargetP and Protcomp bioinformatics methods were used to analysis the B. bassiana genome that contain 10 364 protein,totally 940 secreted proteins were obtained,including 185 carbohydrate active enzyme proteins. Among them,there are 40 carbohydrate binding modules,23 carbohydrate esterases,92 glycoside hydrolases,4 glycosyltransferases,2 polysaccharides lysases,and 24 auxiliary activities. Furtherly the above secretory proteins were blast with the extracellular enzyme database the cysteine content and multiple tandem repeats were analyzed. Compared with the Pathogen-Host interaction database ant then screened out the proteins larger than 300 amino acid. Totally 18 proteins were confirmed to be the effectors of B. bassiana. This study used bioinformatics to predict candidate effectors from B. bassiana genomic,laid a foundation for further research on interaction between pathogen and insect host,and provide reference for the prediction and analysis of other pathogen effectors.
引文
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[2] Valerojiménez C A,Wiegers H,Zwaan B J,Koenraadt C J M,Kan J A LⅤ. Genes involved in virulence of the entomopathogenic fungus Beauveria bassiana[J]. Journal of Invertebrate Pathology,2016,133(1):41-49
[3] Ali G,Wang B,Cao N,Lin H F. Pathogenicity of Beauveria bassiana strain 202 against sap-sucking insect pests[J]. Plant Protection Science,2017,54(1):1-7
[4]季香云,杨长举.白僵菌的致病性与应用[J].中国生物防治,2003,19(2):82-85
[5]李春香,张淑红.白僵菌对害虫致病性的研究进展[J].唐山师范学院学报,2005,27(5):42-43
[6] Wang C,Fan M,Li Z,Butt T M. Molecular monitoring and evaluation of the application of the insect-pathogenic fungus Beauveria bassiana in southeast China[J]. Journal of Applied Microbiology,2004,96(4):861-870
[7] Li Z Z,Alves SB,Roberts D W,Fan M Z,Delalibera Júnior I,Jian T. Biological control of insects in brazil and china:history,current programs and reasons for their successes using entomopathogenic fungi[J]. Biocontrol Science&Technology,2010,20(2):117-136
[8] Xiao G,Ying S H,Peng Z,Wang Z L,Zhang S,Xie X Q. Genomic perspectives on the evolution of fungal entomopathogenicity in Beauveria bassiana[J]. Scientific Reports,2012,2(483):483
[9] Cooper B,Campbell K B,Beard H S,Garrett W M,Islam N.Putative rust fungal effector proteins in infected bean and soybean leaves[J]. Phytopathology,2016,106(5):491-499
[10] Selin C, Kievit T R D, Belmonte M F, Fernando W G D.Elucidating the role of effectors in plant-fungal interactions:progress and challenges[J]. Frontiers in Microbiology,2016,7(967):1-21
[11] Ramachandran S R,Yin C,Kud J,Tanaka K,Mahoney A K,Xiao F. Effectors from wheat rust fungi suppress multiple plant defense responses[J]. Phytopathology,2017,107(1):75
[12] Tan K C,Oliver R P. Regulation of proteinaceous effector expression in phytopathogenic fungi[J]. PLoS Pathogens, 2017, 13(4):e1006241
[13] Van D W A P,Howlett B J. Fungal pathogenicity genes in the age of‘omics’[J]. Molecular Plant Pathology,2011,12(5):507
[14] Jones J D,Dangl J L. The plant immune system[J]. Nature,2006,444(7117):323-329
[15] Stergiopoulos I,de Wit P J. Fungal effector proteins[J]. Annual Review of Phytopathology,2009,47(1):233-263
[16] Bolton M D,Van Esse H P,Vossen J H,De J R,Stergiopoulos I,Stulemeijer I J. The novel Cladosporium fulvum lysin motif effector Ecp6 is a virulence factor with orthologues in other fungal species[J]. Molecular Microbiology,2008,69(1):119-136
[17]闫丽斌,肖淑芹,薛春生.玉米大斑病菌全基因组候选效应分子的预测和分析[J].沈阳农业大学学报,2017,48(1):15-20
[18] Yoshida K,Saitoh H,Fujisawa S,Kanzaki H,Matsumura H,Yoshida K. Association genetics reveals three novel avirulence genes from the rice blast fungal pathogen Magnaporthe oryzae[J]. Plant Cell,2009,21(5):1573
[19]韩长志.全基因组预测禾谷炭疽菌的分泌蛋白[J].生物技术,2014,24(2):36-41
[20]陈琦光,王陈骄子,杨媚,周而勋.希金斯刺盘孢全基因组候选效应分子的预测[J].热带作物学报,2015,36(6):1105-1111
[21] Liu T L,Song T Q,Zhang X, Yuan H, Su L, Li W.Unconventionally secreted effectors of two filamentous pathogens target plant salicylatebiosynthesis[J]. Nature Communications,2014,5(1):4686
[22] Sonah H,Deshmukh R K,Bélanger R R. Computational prediction of effector proteins in fungi:opportunities and challenges[J].Frontiers in Plant Science,2016,7(3):126
[23] Badouin H,Gladieux P,Gouzy J,Siguenza S,Aguileta G,Snirc A.Widespread selective sweeps throughout the genome of model plantpathogenic fungi and identification of effector candidates[J].Molecular Ecology,2017,26(7):2041
[24] Soyer J L, Ghalid M E, Glaser N, Ollivier B, Linglin J,Grandaubert J. Epigenetic control of effector gene expression in the plant pathogenic fungus Leptosphaeria maculans[J]. PLoS Genetics,2014,10(3):e1004227
[25] Wang C,Zhang S,Hou R,Zhao Z,Zheng Q,Xu Q. Functional analysis of the kinome of the wheat scab fungus Fusarium graminearum[J]. PLoS Pathogens,2011,7(12):e1002460
[26] Dor S,Kinch L N,Trudgian D C,Xiao F G,Klimko J A,Grishin N V. Marker for typeⅥsecretion system effectors[J]. Proceedings of the National Academy of Sciences of the United States of America,2014,111(25):9271-9276
[27] Dong Y,Morton J J,Ramirez J L,Souzaneto J A,Dimopoulos G.The entomopathogenic fungus Beauveria bassiana activate toll and JAK-STAT pathway-controlled effector genes and anti-dengue activity in Aedes aegypti[J]. Insect Biochemistry&Molecular Biology,2012,42(2):126-132
[28] Cen K,Li B,Lu Y,Zhang S,Wang C. Divergent LysMeffectors contribute to the virulence of Beauveria bassiana by evasion of insect immune defenses.[J]. PLoS Pathogens,2017,13(9):e1006604
[29]周晓罡,侯思名,陈铎文,陶南,丁玉梅,孙茂林,张绍松.马铃薯晚疫病菌全基因组分泌蛋白的初步分析[J].遗传,2011,33(7):785-793
[30]陈琦光,王陈骄子,杨媚,周而勋.希金斯刺盘孢全基因组候选效应分子的预测[J].热带作物学报,2015,36(6):1105-1111