In this study,Hy322 gene was cloned from Vibrio alginolyticus.The total length of its gene was 969 bp,and it could encode 322 amino acids.The physicochemical properties,protein structure,genetic evolutionary relations...In this study,Hy322 gene was cloned from Vibrio alginolyticus.The total length of its gene was 969 bp,and it could encode 322 amino acids.The physicochemical properties,protein structure,genetic evolutionary relationship and antigenic characteristics of the effector protein Hy322 of V.alginolyticus HY9901 type Ⅲ secretion system were studied and analyzed by bioinformatics methods and tools.The results showed that Hy322 is an unstable hydrophilic and acidic protein without a transmembrane region and a signal peptide,and secondary structure to α-helix.The evolutionary analysis showed that V.alginolyticus HY9901 and V.harveyi were clustered together,which indicated that the genetic relationship between the two species was closest.HY322 contains a FliN super family conserved domain associated with Flagellar motor switch.Bioinformatics analysis showed that the B-cell preponderant epitopes of Hy322 might be localized in the regions of 32-33,100-102,138-140,215-216,235-238 and 246-249.The 3D structure model of Hy322 subunit was simulated by SWISS-MODEL software and itwas found that the yscQ of Yersinia were similar and the similarity was 42.25%.In this study,the feasibility of Hy322 as a common antigen of Vibrio was verified from the perspective of bioinformatics,which laid the foundation for the next step in vaccine development.展开更多
Peanut(Arachis hypogaea),which is widely cultivated across the world,provides high-quality vegetable oil,protein,dietary fiber,minerals,and vitamins for humans.However,in field conditions,the peanut is easily affected...Peanut(Arachis hypogaea),which is widely cultivated across the world,provides high-quality vegetable oil,protein,dietary fiber,minerals,and vitamins for humans.However,in field conditions,the peanut is easily affected by various biotic and abiotic stresses.Diplodia gossypina is the dominant pathogen causing severe collar rot on peanuts.To dissect the pathogenic mechanism of D.gossypina,genome sequencing analysis was performed by using the D.gossypina strain A20_4.The sequencing data showed that the genome assembly size of D.gossypina A20_4 is 43.03 Mb with a GC content of 54.91%.The de novo assembly identified a total of 10,745 genes,containing 41,526 coding sequences and 2.20%of repeat sequences,of which 6,461 genes(60.13%)were annotated using BlastP from GO annotation,3,245 genes(30.20%)and 3,093 genes(28.79%)were annotated from KOG and KEGG annotations,respectively.Meanwhile,the secreted proteins and effectors in 10,745 protein sequences encoded by the whole genome of D.gossypina A20_4 were analyzed,and the results showed that there are 790 secreted protein genes including 220 carbohydrate-active enzymes and 224 potential effector proteins.The functions of 222 potential effector proteins can be annotated by PHI-base.According to the annotation results,12 key pathogenic factors were identified in D.gossypina A20_4.Moreover,a serine/threonine protein kinase SNF1 gene required for autophagy process was identified and analyzed.Deciphering the whole genome of D.gossypina A20_4 provides us with novel insights into understanding evolution,pathogenic molecular mechanism,host-pathogen interaction,and many other complexities of the pathogen.展开更多
【目的】明确山田胶锈菌(Gymnosporangium yamadae)吸器中效应蛋白GyHGSRE1的基础生物学功能,为进一步揭示山田胶锈菌效应蛋白作用的分子机制,制定持久的锈病防控策略奠定基础。【方法】基于前期获得的山田胶锈菌吸器转录组数据筛选具...【目的】明确山田胶锈菌(Gymnosporangium yamadae)吸器中效应蛋白GyHGSRE1的基础生物学功能,为进一步揭示山田胶锈菌效应蛋白作用的分子机制,制定持久的锈病防控策略奠定基础。【方法】基于前期获得的山田胶锈菌吸器转录组数据筛选具有高表达丰度的效应蛋白;通过MEME(http://meme-suite.org/)、AI(https://drug.ai.tencent.com)等在线网站预测GyHGSRE1的蛋白质结构;利用荧光定量PCR检测GyHGSRE1基因在山田胶锈菌侵染过程中的表达水平,以酵母菌分泌系统验证GyHGSRE1信号肽的分泌功能,并通过农杆菌介导的瞬时表达技术分析GyHGSRE1在本氏烟(Nicotiana benthamiana)和苹果(Malus domestica)叶片中的功能。【结果】在山田胶锈菌吸器转录组数据中获得了FPKM(fragments per Kb per million mapped reads)值为117.92、富含甘氨酸和丝氨酸的小分子分泌蛋白GyHGSRE1。其N端具有1个富含丝氨酸motif的信号肽。qRT-PCR显示GyHGSRE1基因在山田胶锈菌的吸器形成阶段以及性、锈孢子形成及发育阶段均上调表达,并定位于本氏烟叶片细胞的细胞质和细胞核,能够诱导细胞坏死并激发基础免疫防御反应,但含有信号肽的全长效应蛋白GyHGSRE1能够诱导苹果叶片细胞坏死,去除信号肽后诱导细胞坏死的能力减弱。【结论】富含甘氨酸和丝氨酸的非典型效应蛋白GyHGSRE1定位于植物细胞的细胞质和细胞核,能够诱导本氏烟及苹果叶片细胞坏死,因此可能具有广谱性的激发子活性。GyHGSRE1在山田胶锈菌侵染苹果叶片的定殖和孢子发育阶段显著表达从而发挥功能,其信号肽可能决定着对寄主特异性识别的相关功能。展开更多
基金Supported by Shenzhen Science and Technology Project(JCYJ20170818111629778,JCYJ20170306161613251)National Natural Science Foundation of Guangdong Province(2017A030313174)+1 种基金Natural Science Foundation of Guangdong Ocean University(C17379)Undergraduate Innovative and Entrepreneurial Team Project(CCTD201802)
文摘In this study,Hy322 gene was cloned from Vibrio alginolyticus.The total length of its gene was 969 bp,and it could encode 322 amino acids.The physicochemical properties,protein structure,genetic evolutionary relationship and antigenic characteristics of the effector protein Hy322 of V.alginolyticus HY9901 type Ⅲ secretion system were studied and analyzed by bioinformatics methods and tools.The results showed that Hy322 is an unstable hydrophilic and acidic protein without a transmembrane region and a signal peptide,and secondary structure to α-helix.The evolutionary analysis showed that V.alginolyticus HY9901 and V.harveyi were clustered together,which indicated that the genetic relationship between the two species was closest.HY322 contains a FliN super family conserved domain associated with Flagellar motor switch.Bioinformatics analysis showed that the B-cell preponderant epitopes of Hy322 might be localized in the regions of 32-33,100-102,138-140,215-216,235-238 and 246-249.The 3D structure model of Hy322 subunit was simulated by SWISS-MODEL software and itwas found that the yscQ of Yersinia were similar and the similarity was 42.25%.In this study,the feasibility of Hy322 as a common antigen of Vibrio was verified from the perspective of bioinformatics,which laid the foundation for the next step in vaccine development.
基金The Major Science and Technology Project of Henan Province(Grant No.201300111000)The Basic scientific Research Project of Henan Academy of Agricultural Sciences(Grant No.2022ZC37,2023ZC46)。
文摘Peanut(Arachis hypogaea),which is widely cultivated across the world,provides high-quality vegetable oil,protein,dietary fiber,minerals,and vitamins for humans.However,in field conditions,the peanut is easily affected by various biotic and abiotic stresses.Diplodia gossypina is the dominant pathogen causing severe collar rot on peanuts.To dissect the pathogenic mechanism of D.gossypina,genome sequencing analysis was performed by using the D.gossypina strain A20_4.The sequencing data showed that the genome assembly size of D.gossypina A20_4 is 43.03 Mb with a GC content of 54.91%.The de novo assembly identified a total of 10,745 genes,containing 41,526 coding sequences and 2.20%of repeat sequences,of which 6,461 genes(60.13%)were annotated using BlastP from GO annotation,3,245 genes(30.20%)and 3,093 genes(28.79%)were annotated from KOG and KEGG annotations,respectively.Meanwhile,the secreted proteins and effectors in 10,745 protein sequences encoded by the whole genome of D.gossypina A20_4 were analyzed,and the results showed that there are 790 secreted protein genes including 220 carbohydrate-active enzymes and 224 potential effector proteins.The functions of 222 potential effector proteins can be annotated by PHI-base.According to the annotation results,12 key pathogenic factors were identified in D.gossypina A20_4.Moreover,a serine/threonine protein kinase SNF1 gene required for autophagy process was identified and analyzed.Deciphering the whole genome of D.gossypina A20_4 provides us with novel insights into understanding evolution,pathogenic molecular mechanism,host-pathogen interaction,and many other complexities of the pathogen.
文摘【目的】明确山田胶锈菌(Gymnosporangium yamadae)吸器中效应蛋白GyHGSRE1的基础生物学功能,为进一步揭示山田胶锈菌效应蛋白作用的分子机制,制定持久的锈病防控策略奠定基础。【方法】基于前期获得的山田胶锈菌吸器转录组数据筛选具有高表达丰度的效应蛋白;通过MEME(http://meme-suite.org/)、AI(https://drug.ai.tencent.com)等在线网站预测GyHGSRE1的蛋白质结构;利用荧光定量PCR检测GyHGSRE1基因在山田胶锈菌侵染过程中的表达水平,以酵母菌分泌系统验证GyHGSRE1信号肽的分泌功能,并通过农杆菌介导的瞬时表达技术分析GyHGSRE1在本氏烟(Nicotiana benthamiana)和苹果(Malus domestica)叶片中的功能。【结果】在山田胶锈菌吸器转录组数据中获得了FPKM(fragments per Kb per million mapped reads)值为117.92、富含甘氨酸和丝氨酸的小分子分泌蛋白GyHGSRE1。其N端具有1个富含丝氨酸motif的信号肽。qRT-PCR显示GyHGSRE1基因在山田胶锈菌的吸器形成阶段以及性、锈孢子形成及发育阶段均上调表达,并定位于本氏烟叶片细胞的细胞质和细胞核,能够诱导细胞坏死并激发基础免疫防御反应,但含有信号肽的全长效应蛋白GyHGSRE1能够诱导苹果叶片细胞坏死,去除信号肽后诱导细胞坏死的能力减弱。【结论】富含甘氨酸和丝氨酸的非典型效应蛋白GyHGSRE1定位于植物细胞的细胞质和细胞核,能够诱导本氏烟及苹果叶片细胞坏死,因此可能具有广谱性的激发子活性。GyHGSRE1在山田胶锈菌侵染苹果叶片的定殖和孢子发育阶段显著表达从而发挥功能,其信号肽可能决定着对寄主特异性识别的相关功能。
