病原真菌通常分泌效应子到寄主组织中调控寄主的生理过程,从而有利于其侵染。CFEM(common in several fungal extracellular membrane)蛋白是真菌所独有的,且与致病性密切相关。本研究利用Pfam数据库对草莓胶孢炭疽菌全基因组进行搜索,...病原真菌通常分泌效应子到寄主组织中调控寄主的生理过程,从而有利于其侵染。CFEM(common in several fungal extracellular membrane)蛋白是真菌所独有的,且与致病性密切相关。本研究利用Pfam数据库对草莓胶孢炭疽菌全基因组进行搜索,鉴定获得22个CFEM蛋白。对CFEM蛋白的信号肽、跨膜结构域和亚细胞定位进行分析,结果表明仅有8个CFEM蛋白为分泌蛋白。对CFEM分泌蛋白在不同侵染阶段的转录情况进行转录组学及RT-PCR分析,结果显示8个CFEM蛋白在侵染后不同时期均有表达。其中,1个CFEM分泌蛋白于附着胞形成期特异表达,2个于活体寄生阶段特异表达,2个于死体寄生阶段特异表达。综合上述分析结果,预测这8个分泌蛋白可能为草莓胶孢炭疽菌的效应子。本研究为深入解析植物病原真菌CFEM效应子提供了理论依据。展开更多
[目的]本文旨在研究辣椒胶孢炭疽菌(Colletotrichum gloeosporioides)常见真菌细胞外膜蛋白(common in several fungal extracellular membrane proteins,CFEM)的组成和结构。[方法]以辣椒胶孢炭疽菌CSLL^(-1)1的基因组为研究对象,通过...[目的]本文旨在研究辣椒胶孢炭疽菌(Colletotrichum gloeosporioides)常见真菌细胞外膜蛋白(common in several fungal extracellular membrane proteins,CFEM)的组成和结构。[方法]以辣椒胶孢炭疽菌CSLL^(-1)1的基因组为研究对象,通过生物信息学分析其CFEM效应因子的信号肽、跨膜结构域和亚细胞定位信号等结构特征;利用转录组学途径分析其在病菌与寄主互作过程中的表达特征,并对部分CFEM效应因子进行亚细胞定位分析。[结果]CSLL^(-1)1共编码30个CFEM效应因子,其中26个具有信号肽并可分泌至胞外;17个含有数目不等的跨膜结构域。CFEM效应因子Cghn02929、Cghn09727、Cghn12645和Cghn14146在侵染辣椒果实各阶段均具有较高表达水平,推测其可能参与病菌整个侵染过程。Cghn14146、Cghn09727和Cghn13279定位于细胞膜和细胞质,而Cghn02929、Cghn13471定位于细胞核、细胞膜和细胞质。[结论]明确了辣椒胶孢炭疽菌CFEM效应因子的组成和结构特征,探明了CFEM效应因子间的表达水平和亚细胞定位差异。展开更多
植物病原真菌效应子是指可以改变寄主植物细胞结构或者细胞功能的分泌蛋白或其他小分子物质。效应子对病原真菌的侵入、扩展以及致病发挥着至关重要的作用,是植物病原真菌与寄主的互作不断演化的必然结果。真菌特有的CFEM(common in sev...植物病原真菌效应子是指可以改变寄主植物细胞结构或者细胞功能的分泌蛋白或其他小分子物质。效应子对病原真菌的侵入、扩展以及致病发挥着至关重要的作用,是植物病原真菌与寄主的互作不断演化的必然结果。真菌特有的CFEM(common in several fungal extracellular membrane protein)蛋白对于病原真菌的致病性起重要作用,一些能够被分泌到胞外的CFEM蛋白被证明是病原真菌效应子。由禾谷炭疽菌(Colletotrichum graminicola)引起的玉米炭疽病是玉米上的重要病害,常年造成严重损失。本研究运用生物信息学工具对禾谷炭疽菌中的CFEM蛋白进行信号肽分析和亚细胞定位分析,进而通过转录分析明确禾谷炭疽菌CFEM蛋白的表达时期。分析结果表明,该病原真菌编码32个CFEM蛋白,其中22个具有信号肽并可分泌至胞外,为分泌蛋白。转录分析表明,10个CFEM分泌蛋白于病菌侵染时附着胞形成期表达,2个CFEM分泌蛋白于侵染后的活体寄生阶段表达,1个于死体寄生阶段表达,其余9个CFEM分泌蛋白在病菌侵染时期的3个阶段均稳定表达。结合生物信息学和转录分析结果,我们预测这22个CFEM分泌蛋白为禾谷炭疽菌致病相关的效应子(简称CFEM效应子)。明确禾谷炭疽菌中CFEM蛋白数量,预测病菌致病相关的CFEM效应子组成,可为开展病原真菌CFEM蛋白介导的病菌—寄主互作研究奠定基础,并为玉米炭疽病的防治和抗性育种研究提供参考。展开更多
病原真菌在侵染植物过程中,往往分泌效应因子增加致病性或被寄主抗病基因识别激发寄主强烈抗性,因此效应因子在植物-病原物的互作中发挥重要作用。CFEM(Common in Fungal Extracellular Membrane)蛋白是真菌中特有的一类位于细胞外膜的...病原真菌在侵染植物过程中,往往分泌效应因子增加致病性或被寄主抗病基因识别激发寄主强烈抗性,因此效应因子在植物-病原物的互作中发挥重要作用。CFEM(Common in Fungal Extracellular Membrane)蛋白是真菌中特有的一类位于细胞外膜的蛋白,往往起效应因子的作用。本文综述了植物病原真菌CFEM蛋白家族的结构特点,在不同物种中的表达和定位,CFEM蛋白对真菌胞内铁吸收和生长发育方面的调控作用,抑制寄主免疫反应和促进真菌寄生的作用机制以及CFEM蛋白起源和进化过程等方面的研究进展,同时对目前CFEM蛋白研究有待阐明的问题进行了讨论并展望了未来的研究方向。本文将有助于人们进一步了解植物-病原真菌互作分子机理以及CFEM蛋白的致病机理,为培育作物抗病品种和制定植物真菌病害防控策略提供参考。展开更多
Fungal secreted proteins that contain the Common in Fungal Extracellular Membrane(CFEM)domain are important for pathogenicity.The hemibiotrophic fungus Colletotrichum graminicola causes the serious anthracnose disease...Fungal secreted proteins that contain the Common in Fungal Extracellular Membrane(CFEM)domain are important for pathogenicity.The hemibiotrophic fungus Colletotrichum graminicola causes the serious anthracnose disease of maize.In this study,we identified 24 CgCFEM proteins in the genome of C.graminicola.Phylogenic analysis revealed that these 24 proteins(CgCFEM1–24)can be divided into 2 clades based on the presence of the trans-membrane domain.Sequence alignment analysis indicated that the amino acids of the CFEM domain are highly conserved and contain 8 spaced cysteines,with the exception that CgCFEM1 and CgCFEM24 lack 1 and 2 cysteines,respectively.Ten CgCFEM proteins with a signal peptide and without the trans-membrane domain were considered as candidate effectors and,thus were selected for structural prediction and functional analyses.The CFEM domain in the candidate effectors can form a helical-basket structure homologous to the Csa2 protein in Candida albicans,which is responsible for haem acquisition and pathogenicity.Subcellular localization analysis revealed that these effectors accumulate in the cell membrane,nucleus,and cytosolic bodies.Additionally,5 effectors,CgCFEM6,7,8,9 and 15,can suppress the BAX-induced programmed cell death in Nicotiana benthamiana with or without the signal peptide.These results demonstrate that these 10 CgCFEM candidate effectors with different structures and subcellular localizations in host cells may play important roles during the pathogenic processes on maize plants.展开更多
Common in Fungal Extracellular Membrane(CFEM)domains are uniquely found in fungal extracellular membrane proteins which are important for pathogens.This study identified 13 StCFEM proteins in the genome of Setosphaeri...Common in Fungal Extracellular Membrane(CFEM)domains are uniquely found in fungal extracellular membrane proteins which are important for pathogens.This study identified 13 StCFEM proteins in the genome of Setosphaeria turcica,the hemibiotrophic fungus that causes northern corn leaf blight.Sequence alignment and WebLogo analysis of their CFEM domains indicated that the amino acids were highly conserved and that,with the exception of StCFEM1,2,3,and 6,they contained eight cysteines.Phylogenic analysis suggested that these 13 proteins(StCFEM1–13)could be divided into 2 clades based on the presence of the trans-membrane domain.Six StCFEM proteins with a signal peptide and without a trans-membrane domain were considered as candidate effector proteins.The CFEM domain in the candidate effector proteins could form a helical-basket structure homologous to Csa2 in Candida albicans.Transcriptome analysis suggested that the 13 genes were expressed during fungal infection and a yeast secretion assay revealed that these candidate effectors were secreted proteins.It was also found that StCFEM3,4,and 5 couldn't affect BAX/INF1-induced programmed cell death(PCD)in Nicotiana benthamiana and while StCFEM12 could suppress INF1-induced PCD,it showed no effect on BAXinduced PCD.This study found that there were 13 members of the S.turcica CFEM protein family and that StCFEM12 was a candidate effector.This study laid the foundation for illustrating the roles of CFEM proteins during the pathogenic processes of phytopathogens.展开更多
Common in Fungal Extracellular Membrane(CFEM)domain proteins are considered to be unique to fungi and closely related to pathogenicity.However,the Puccinia striiformis f.sp.tritici(Pst)effector containing the CFEM dom...Common in Fungal Extracellular Membrane(CFEM)domain proteins are considered to be unique to fungi and closely related to pathogenicity.However,the Puccinia striiformis f.sp.tritici(Pst)effector containing the CFEM domain has not been reported.Here,we obtained an effector,PstCFEM1,containing a functional N-terminal signal peptide sequence and the CFEM domain from Pst race CYR31.qRT-PCR assay indicated that the transcript levels of PstCFEM1 were highly induced during the early stages of infection.Overexpression of PstCFEM1 suppressed Pst322(an elicitor-like protein of Pst)-trigged cell death,reactive oxygen species(ROS)accumulation and callose deposition.Host-induced gene silencing(HIGS)experiments showed that knockdown of PstCFEM1 decreased the virulence of Pst,while ROS accumulation in silenced plants increased near the infection site.In addition,wheat containing the PstCFEM1-silenced construct increased resistance to multiple races of Pst.Our data suggest that PstCFEM1 suppresses wheat defense by inhibiting ROS accumulation and contributes to increased virulence of Pst.展开更多
A new method, the characteristic finite element method （CFEM）, was developed to simulate solute transport in a cross-fracture. The solution of this mathematical model for solute transport considered that the contrib...A new method, the characteristic finite element method （CFEM）, was developed to simulate solute transport in a cross-fracture. The solution of this mathematical model for solute transport considered that the contribution of convection and dispersion terms was deduced using the single-step, trace-back method and routine finite element method （FEM）. Also, experimental models were designed to verify the reliability and validity of the CFEM. Results showed that experimental data from a single fracture model agreed with numerical simulations obtained from the use of the CFEM. However, routine FEM caused numerical oscillation and dispersion during the calculation of solute concentration. Furthermore, in this cross-fracture model, CFEM simulation results predicted that the arrival time of concentration peak values decreased with increasing flux. Also, the second concentration peak value was obvious with the decrease of flux, which may have resulted from the convergence of solute concentrations from main, and branch, fractures.展开更多
文摘病原真菌通常分泌效应子到寄主组织中调控寄主的生理过程,从而有利于其侵染。CFEM(common in several fungal extracellular membrane)蛋白是真菌所独有的,且与致病性密切相关。本研究利用Pfam数据库对草莓胶孢炭疽菌全基因组进行搜索,鉴定获得22个CFEM蛋白。对CFEM蛋白的信号肽、跨膜结构域和亚细胞定位进行分析,结果表明仅有8个CFEM蛋白为分泌蛋白。对CFEM分泌蛋白在不同侵染阶段的转录情况进行转录组学及RT-PCR分析,结果显示8个CFEM蛋白在侵染后不同时期均有表达。其中,1个CFEM分泌蛋白于附着胞形成期特异表达,2个于活体寄生阶段特异表达,2个于死体寄生阶段特异表达。综合上述分析结果,预测这8个分泌蛋白可能为草莓胶孢炭疽菌的效应子。本研究为深入解析植物病原真菌CFEM效应子提供了理论依据。
文摘[目的]本文旨在研究辣椒胶孢炭疽菌(Colletotrichum gloeosporioides)常见真菌细胞外膜蛋白(common in several fungal extracellular membrane proteins,CFEM)的组成和结构。[方法]以辣椒胶孢炭疽菌CSLL^(-1)1的基因组为研究对象,通过生物信息学分析其CFEM效应因子的信号肽、跨膜结构域和亚细胞定位信号等结构特征;利用转录组学途径分析其在病菌与寄主互作过程中的表达特征,并对部分CFEM效应因子进行亚细胞定位分析。[结果]CSLL^(-1)1共编码30个CFEM效应因子,其中26个具有信号肽并可分泌至胞外;17个含有数目不等的跨膜结构域。CFEM效应因子Cghn02929、Cghn09727、Cghn12645和Cghn14146在侵染辣椒果实各阶段均具有较高表达水平,推测其可能参与病菌整个侵染过程。Cghn14146、Cghn09727和Cghn13279定位于细胞膜和细胞质,而Cghn02929、Cghn13471定位于细胞核、细胞膜和细胞质。[结论]明确了辣椒胶孢炭疽菌CFEM效应因子的组成和结构特征,探明了CFEM效应因子间的表达水平和亚细胞定位差异。
文摘植物病原真菌效应子是指可以改变寄主植物细胞结构或者细胞功能的分泌蛋白或其他小分子物质。效应子对病原真菌的侵入、扩展以及致病发挥着至关重要的作用,是植物病原真菌与寄主的互作不断演化的必然结果。真菌特有的CFEM(common in several fungal extracellular membrane protein)蛋白对于病原真菌的致病性起重要作用,一些能够被分泌到胞外的CFEM蛋白被证明是病原真菌效应子。由禾谷炭疽菌(Colletotrichum graminicola)引起的玉米炭疽病是玉米上的重要病害,常年造成严重损失。本研究运用生物信息学工具对禾谷炭疽菌中的CFEM蛋白进行信号肽分析和亚细胞定位分析,进而通过转录分析明确禾谷炭疽菌CFEM蛋白的表达时期。分析结果表明,该病原真菌编码32个CFEM蛋白,其中22个具有信号肽并可分泌至胞外,为分泌蛋白。转录分析表明,10个CFEM分泌蛋白于病菌侵染时附着胞形成期表达,2个CFEM分泌蛋白于侵染后的活体寄生阶段表达,1个于死体寄生阶段表达,其余9个CFEM分泌蛋白在病菌侵染时期的3个阶段均稳定表达。结合生物信息学和转录分析结果,我们预测这22个CFEM分泌蛋白为禾谷炭疽菌致病相关的效应子(简称CFEM效应子)。明确禾谷炭疽菌中CFEM蛋白数量,预测病菌致病相关的CFEM效应子组成,可为开展病原真菌CFEM蛋白介导的病菌—寄主互作研究奠定基础,并为玉米炭疽病的防治和抗性育种研究提供参考。
文摘病原真菌在侵染植物过程中,往往分泌效应因子增加致病性或被寄主抗病基因识别激发寄主强烈抗性,因此效应因子在植物-病原物的互作中发挥重要作用。CFEM(Common in Fungal Extracellular Membrane)蛋白是真菌中特有的一类位于细胞外膜的蛋白,往往起效应因子的作用。本文综述了植物病原真菌CFEM蛋白家族的结构特点,在不同物种中的表达和定位,CFEM蛋白对真菌胞内铁吸收和生长发育方面的调控作用,抑制寄主免疫反应和促进真菌寄生的作用机制以及CFEM蛋白起源和进化过程等方面的研究进展,同时对目前CFEM蛋白研究有待阐明的问题进行了讨论并展望了未来的研究方向。本文将有助于人们进一步了解植物-病原真菌互作分子机理以及CFEM蛋白的致病机理,为培育作物抗病品种和制定植物真菌病害防控策略提供参考。
基金supported by the National Program for Support of Top-notch Young Professionals of China
文摘Fungal secreted proteins that contain the Common in Fungal Extracellular Membrane(CFEM)domain are important for pathogenicity.The hemibiotrophic fungus Colletotrichum graminicola causes the serious anthracnose disease of maize.In this study,we identified 24 CgCFEM proteins in the genome of C.graminicola.Phylogenic analysis revealed that these 24 proteins(CgCFEM1–24)can be divided into 2 clades based on the presence of the trans-membrane domain.Sequence alignment analysis indicated that the amino acids of the CFEM domain are highly conserved and contain 8 spaced cysteines,with the exception that CgCFEM1 and CgCFEM24 lack 1 and 2 cysteines,respectively.Ten CgCFEM proteins with a signal peptide and without the trans-membrane domain were considered as candidate effectors and,thus were selected for structural prediction and functional analyses.The CFEM domain in the candidate effectors can form a helical-basket structure homologous to the Csa2 protein in Candida albicans,which is responsible for haem acquisition and pathogenicity.Subcellular localization analysis revealed that these effectors accumulate in the cell membrane,nucleus,and cytosolic bodies.Additionally,5 effectors,CgCFEM6,7,8,9 and 15,can suppress the BAX-induced programmed cell death in Nicotiana benthamiana with or without the signal peptide.These results demonstrate that these 10 CgCFEM candidate effectors with different structures and subcellular localizations in host cells may play important roles during the pathogenic processes on maize plants.
基金supported by the grants from the Natural Science Foundation of Hebei Province,China(C2020204172 and C2018204120)the China Agriculture Research System of MOF and MARA(CARS-02-25)the National Natural Science Foundation of China(31601598 and 31901827)。
文摘Common in Fungal Extracellular Membrane(CFEM)domains are uniquely found in fungal extracellular membrane proteins which are important for pathogens.This study identified 13 StCFEM proteins in the genome of Setosphaeria turcica,the hemibiotrophic fungus that causes northern corn leaf blight.Sequence alignment and WebLogo analysis of their CFEM domains indicated that the amino acids were highly conserved and that,with the exception of StCFEM1,2,3,and 6,they contained eight cysteines.Phylogenic analysis suggested that these 13 proteins(StCFEM1–13)could be divided into 2 clades based on the presence of the trans-membrane domain.Six StCFEM proteins with a signal peptide and without a trans-membrane domain were considered as candidate effector proteins.The CFEM domain in the candidate effector proteins could form a helical-basket structure homologous to Csa2 in Candida albicans.Transcriptome analysis suggested that the 13 genes were expressed during fungal infection and a yeast secretion assay revealed that these candidate effectors were secreted proteins.It was also found that StCFEM3,4,and 5 couldn't affect BAX/INF1-induced programmed cell death(PCD)in Nicotiana benthamiana and while StCFEM12 could suppress INF1-induced PCD,it showed no effect on BAXinduced PCD.This study found that there were 13 members of the S.turcica CFEM protein family and that StCFEM12 was a candidate effector.This study laid the foundation for illustrating the roles of CFEM proteins during the pathogenic processes of phytopathogens.
基金supported by the National Key Research and Development Program of China(2021YFD1401003)the National Natural Science Foundation of China(32172381 and 31972224),Key Research and Development Program of Shaanxi(2021ZDLNY01–01)+1 种基金Natural Science Basic Research Program of Shaanxi(2020JZ-13)the 111 Project from the Ministry of Education of China(B07049).
文摘Common in Fungal Extracellular Membrane(CFEM)domain proteins are considered to be unique to fungi and closely related to pathogenicity.However,the Puccinia striiformis f.sp.tritici(Pst)effector containing the CFEM domain has not been reported.Here,we obtained an effector,PstCFEM1,containing a functional N-terminal signal peptide sequence and the CFEM domain from Pst race CYR31.qRT-PCR assay indicated that the transcript levels of PstCFEM1 were highly induced during the early stages of infection.Overexpression of PstCFEM1 suppressed Pst322(an elicitor-like protein of Pst)-trigged cell death,reactive oxygen species(ROS)accumulation and callose deposition.Host-induced gene silencing(HIGS)experiments showed that knockdown of PstCFEM1 decreased the virulence of Pst,while ROS accumulation in silenced plants increased near the infection site.In addition,wheat containing the PstCFEM1-silenced construct increased resistance to multiple races of Pst.Our data suggest that PstCFEM1 suppresses wheat defense by inhibiting ROS accumulation and contributes to increased virulence of Pst.
基金financially supported by the Project Foundation of Chongqing Municipal Education Committee(KJ131106)The National Basic Research Programme of China(973 Programme,Grant No.2012CB719804)The Special Fund of the State Key Laboratory of Hydrology-Water Resources and Hydraulic Engineering(Grant No.20145027312)
文摘A new method, the characteristic finite element method （CFEM）, was developed to simulate solute transport in a cross-fracture. The solution of this mathematical model for solute transport considered that the contribution of convection and dispersion terms was deduced using the single-step, trace-back method and routine finite element method （FEM）. Also, experimental models were designed to verify the reliability and validity of the CFEM. Results showed that experimental data from a single fracture model agreed with numerical simulations obtained from the use of the CFEM. However, routine FEM caused numerical oscillation and dispersion during the calculation of solute concentration. Furthermore, in this cross-fracture model, CFEM simulation results predicted that the arrival time of concentration peak values decreased with increasing flux. Also, the second concentration peak value was obvious with the decrease of flux, which may have resulted from the convergence of solute concentrations from main, and branch, fractures.
