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Two NIS1-like proteins from apple canker pathogen(Valsa mali)play distinct roles in plant recognition and pathogen virulence

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Stress Biology 2022年第1期2卷 606-620页

作者： jiajun nie Wenjing Zhou Yonghui Lin Zhaoyang Liu Zhiyuan Yin Lili Huang State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A&F University3 Taicheng RoadYangling 712100ShaanxiChina College of Plant Protection China Agricultural UniversityBeijing 100193China

Conserved effectors produced by phytopathogens play critical roles in plant-microbe ***1-like proteins represent a newly identified family of effectors distributed in multiple fungal ***,their biological functions in a majority of pathogenic fungi remain largely elusive and require further *** this study,we characterized two NIS1-like proteins VmNIS1 and VmNIS2 from Valsa mali,the causal agent of apple Valsa *** of these two proteins were predicted to be *** agroinfiltration,we found that VmNIS1 induced intense cell death,whereas VmNIS2 suppressed INF1 elicitin-triggered cell death in Nicotiana *** of *** with VmNIS1 recombinant protein produced by Escherichia coli activated a series of immune responses and enhanced plant disease resistance against Phytophthora *** contrast,VmNIS2 suppressed plant immune responses and promoted *** infection when transiently expressed in *** VmNIS1 and VmNIS2 were shown to be highly induced at late stage of *** *** individually knocking out of these two genes in ***,however,only VmNIS2 was shown to be required for pathogen virulence as well as tolerance to oxidative ***,we further showed that C-terminal extension of VmNIS1 was essential for plant recognition and VmNIS2 may escape plant detection via sequence *** data collectively indicate that VmNIS1 and VmNIS2 play distinct roles in plant recognition and pathogen virulence,which provided new insights into the function of NIS1-like proteins in plant-microbe interactions.

关键词： Valsa mali Effector protein Cell death Plant immunity Virulence

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VmNLP2 from Valsa mali acts as a phytotoxic virulence factor

VmNLP2 from Valsa mali acts as a phytotoxic virulence factor

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中国植物病理学会2021年学术年会

作者： LIU Jianying nie jiajun CHANG Yali HUANG Lili State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A&F University

Necrosis and ethylene-inducing peptide 1(Nepl)-like proteins(NLPs) are typically phytotoxic molecules and play important roles in plant-microbe ***,their biological roles in the pathogenic fungus Valsa mali,the causal agent of apple Valsa canker,remains ***,we identified two NLPs,VmNLPl and VmNLP2 from *** agrobacteriamediated transient expression in Nicotiana benthamiana,only VmNLP2 was shown to induce cell death and was subjected to further *** infiltration of VmNLP2 recombinant protein in apple leaves,we found that VmNLP2 exhibited phytotoxic activity in apple leaves as *** knockout and pathogenicity test showed that VmNLP2-deficient *** strain displayed reduced aggressiveness on *** sequence alignment revealed that the second Histidine(H) among the conserved heptapeptide(GHRHDWE) of VmNLP2 is mutated to Tyrosine(Y).When this Tyrosine(Y) was substituted by Histidine(H),the variant(VmNLP2) showed a stronger cell death 一 inducing activity in ***,complementation of VmNLP2 to VmNLP2 deletion mutant caused larger disease lesion on apple leaves,compared to the *** wild type *** together,we demonstrate that VmNLP2 from *** is an essential virulence factor,and the virulence of VmNLP2 probably correlates to its phytotoxic activity.

关键词： Valsa mali Nep1-like proteins cytotoxicity virulence factor biological roles

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VmE02 from Valsa mali is a new pathogen-associated molecular pattern

VmE02 from Valsa mali is a new pathogen-associated molecular...

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中国植物病理学会第十一届全国会员代表大会暨2018年学术年会

作者： nie jiajun Yin Zhiyuan Li Zhengpeng Wu Yuxing Huang Lili State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A&F University

The necrotrophic fungus Valsa mali causes severe necrosis on apple trees,resulting in substantial yield losses in eastern *** the aim to identify cell death-inducing factors of ***,we carried out transient expression of candidate effector proteins in Nicotiana benthamiana,and VmE02,a protein with unknown function was *** analysis revealed that VmE02 is widely spread in fdamentous *** agroinfiltration,we found there exists homologs of VmE02 in both fungi and oomycetes,which can also elicit cell death in *** of VmE02 in *** activated plant immune responses,including ROS burst,callose deposition,and the expression of both salicylic acid and jasmonate signaling ***,VmE02 induced the expression of PTI marker *** virus-induced gene silencing(VIGS) in ***,we found that cell deathinducing activity of VmE02 requires both Brassinosteroid insensitive 1-Associated Kinase 1(NbBAK1),suppressor of BIR1-1(NbSOBIR1),HSP90,and *** out of VmE02 in *** showed no apparent influence on pathogenicity,however,it considerably reduced pathogen ***,VmE02 manipulated resistance to Sclerotinia sclerotiorum and Phytophthora capsici when expressed in *** conclude that VmE02 is a novel PAMP in fungi and oomycetes,and suggest it is also an effector with diverse function.

关键词： Valsamali Cell death Plant immunity Nicotiana benthamiana Filamentous pathogens Pathogen-associated molecular pattern

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A NIS1-like protein from apple canker pathogen（Valsa mali）may escape plant recognition and serves as a virulence factor

A NIS1-like protein from apple canker pathogen（Valsa mali）...

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中国植物病理学会2021年学术年会

作者： nie jiajun ZHOU Wenjing LIN Yonghui LIU Zhaoyang YIN Zhiyuan HUANG Lili State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A＆F University Department of Plant Pathology China Agricultural University

Phytopathogens-produced conserved effectors play important roles in plant-microbe ***1-like proteins represent a newly identified family of effectors represented in a wide array of fungal ***,their biological roles in pathogenic fungi remain largely *** this study,we characterized two NIS1-like proteins VmNISl and VmNIS2 from Valsa mali,the causal agent of apple Valsa *** found that VmNISl induced intense cell death,whereas VmNIS2 suppressed INF1 elicitin-triggered cell death when transiently expressed in the model plant Nicotiana *** of *** with VmNISl recombinant protein activated a series of immune responses and greatly enhanced plant resistance against Phytophthora *** contrast,VmNIS2 strongly suppressed plant immune responses and promoted the infection of *** when transiently expressed in *** observations indicate that VmNISl but not VmNIS2 can be recognized by *** alignment revealed that,compared to VmNISl,VmNIS2 is truncated at Cterminus by 13 amino acids residues（C13）.Interestingly,deletion of C13 greatly compromised VmNIS1-triggered cell death,whereas fusion of C13 to the C-terminus of VmNIS2 can induce visible cell death in *** indicates that C13 is essential for plant detection of VmNISl,and VmNIS2 may evade plant recognition via C-terminal *** further found that knocking out of VmNIS1 showed no apparent influence on *** virulence,however,knocking out of VmNIS2 obviously attenuated pathogen virulence,suggesting that VmNIS2 is essential for *** infection of *** conclusion,our data suggest that the NIS1-like protein VmNIS2 from the pathogenic *** may have escaped plant recognition and functions as a virulence factor.

关键词： Valsa mali NIS1-like protein cell death immune responses virulence

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热处理对SiC_p/6061Al基复合材料等离子弧焊接头性能的影响

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焊接学报 2008年第11期29卷 9-12页

作者：雷玉成张振陈希章聂加俊江苏大学材料科学与工程学院江苏镇江212013

热处理对SiCp/6061Al基复合材料等离子弧原位焊接接头组织与性能的影响较为明显,固溶处理+时效与退火相比,前者对接头的改善作用更佳。经500℃固溶处理2h+12h时效热处理后,接头组织得到良好的改善,细长的Al3Ti相变为短小棒状,消除了晶... 详细信息

热处理对SiCp/6061Al基复合材料等离子弧原位焊接接头组织与性能的影响较为明显,固溶处理+时效与退火相比,前者对接头的改善作用更佳。经500℃固溶处理2h+12h时效热处理后,接头组织得到良好的改善,细长的Al3Ti相变为短小棒状,消除了晶间偏析,组织更加均匀;热影响区组织中晶粒均得到比较明显的细化,比较接近母材晶粒的大小,焊接接头强度较未热处理时有了较大提高,达到245MPa。

关键词： SiCp/6061A1基复合材料等离子弧热处理显微组织

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Ti—Al—Si对SiCp/Al基复合材料等离子弧焊焊缝的组织与性能的影响

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材料研究学报 2008年第4期22卷 420-424页

作者：雷玉成张振聂加俊陈希章江苏大学材料科学与工程学院镇江212013

以Ti-Al-Si合金作为合金化填充材料,用氮氩混合等离子气体对SiCp/Al基复合材料进行等离子弧原位焊接,研究了Ti-Al-Si对焊缝的组织和性能的影响.结果表明:填加Ti-75Al-5Si合金时,熔池中Si和Ti的联合作用有效抑地制了针状脆生相Al_4C_3的... 详细信息

以Ti-Al-Si合金作为合金化填充材料,用氮氩混合等离子气体对SiCp/Al基复合材料进行等离子弧原位焊接,研究了Ti-Al-Si对焊缝的组织和性能的影响.结果表明:填加Ti-75Al-5Si合金时,熔池中Si和Ti的联合作用有效抑地制了针状脆生相Al_4C_3的生成,形成了稳定的熔池,得到了以TiN、AlN、TiC和Ti_5Si_3等为二次增强相的焊缝.焊缝的组织致密,结合良好,其最大拉伸强度为225 MPa.

关键词：复合材料 SiCp/Al基等离子弧原位焊接 Ti-Al—Si合金

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Ti-Ni合金对SiC_p/AlMMCs等离子弧原位焊接焊缝性能的影响

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焊接学报 2008年第10期29卷 13-16页

作者：雷玉成聂加俊张振陈希章江苏大学材料科学与工程学院江苏镇江212013

以Ti-Ni合金作为填加材料,采用氮氩混合等离子气对SiCp/Al基复合材料进行等离子弧原位焊接。结果表明,填加Ti-Ni合金进行等离子弧原位焊接,在焊缝组织中生成了新的增强颗粒TiN,TiC,AlN,Al3Ti和Al3Ni,未发现明显的针状相生成,有效的抑制... 详细信息

以Ti-Ni合金作为填加材料,采用氮氩混合等离子气对SiCp/Al基复合材料进行等离子弧原位焊接。结果表明,填加Ti-Ni合金进行等离子弧原位焊接,在焊缝组织中生成了新的增强颗粒TiN,TiC,AlN,Al3Ti和Al3Ni,未发现明显的针状相生成,有效的抑制了脆性相Al4C3,并且接头组织致密,结合较好。Ni元素的添加,降低了SiC溶解程度,使得熔池中的结晶热显著提高,改善了熔池的流动性。同时在焊缝组织中形成了新的增强相AlNi,Al3Ni,从而有效地提高接头的力学性能。采用Ti-Ni合金进行等离子弧原位焊接,接头的抗拉强度为215.4 MPa,达到母材强度的67.3%。

关键词： SiCp/Al基复合材料等离子弧原位焊接 Ti-Ni合金

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Identification and characteristic of candidate effectors from Valsa mali

Identification and characteristic of candidate effectors fro...

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中国植物病理学会第十一届全国会员代表大会暨2018年学术年会

作者： Zhang Mian Song Linlin Zhao Yuhuan Zhang Di Li Zhengpeng Yin Zhiyuan nie jiajun Du Hongxia Tan Ni Tian Runze Feng Hao Huang Lili State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A&F University

Valsa mali(***) is the cause of a destructive disease,apple valsa canker,in East *** proteins play important roles in pathogenicity of plant pathogenic fungi in various *** previous study,196 candidate effector proteins(CEPs) were predicted,which were defined as extracellular proteins with no known function and rich in cysteine,and 101 of them were species *** total,184 predicated candidate effector proteins were successfully cloned to PGR106 and 21 candidate effectors were *** the 21 identified candidate effectors(ICEs),there were 18 cell death suppressors(CDS) which accounted for 86%,and three cell death inducers(CDI) which accounted for14%.What is more,2(VmP×E1 and VmEP1) of the 21 identified candidate effectors(ICEs) were identified as important virulence *** result showed that *** possessed affluent and different effectors and evidence also exhibited *** manipulated those effectors to contribute to virulence during infection progress.

关键词： Apple valsa canker Secretome Cell death suppressor Cell death inducer Virulence factor

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苹果黑腐皮壳菌CAP超家族蛋白基因鉴定及毒性功能分析

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中国农业科学 2021年第16期54卷 3440-3450页

作者：王程利尹志远聂嘉俊林永辉黄丽丽西北农林科技大学植物保护学院/旱区作物逆境生物学国家重点实验室

【目的】CAP(Cysteine-rich secretory protein,Antigen 5 and Pathogenesis related protein 1)超家族蛋白广泛存在于真菌、细菌、动植物等物种,并且参与病原菌的致病过程。本研究旨在鉴定苹果树腐烂病致病菌——苹果黑腐皮壳菌(Valsa ... 详细信息

【目的】CAP(Cysteine-rich secretory protein,Antigen 5 and Pathogenesis related protein 1)超家族蛋白广泛存在于真菌、细菌、动植物等物种,并且参与病原菌的致病过程。本研究旨在鉴定苹果树腐烂病致病菌——苹果黑腐皮壳菌(Valsa mali)的CAP超家族基因并明确CAP超家族基因在病菌致病方面的作用。【方法】通过BLASTP在苹果黑腐皮壳菌全基因组中检索具有CAP保守结构域的基因;利用特异性引物对鉴定到的CAP基因进行PCR扩增和凝胶电泳检测;使用生物信息学软件和在线数据库进行蛋白序列特征和系统发育分析;利用RT-qPCR分析基因在病菌侵染过程中的表达模式;使用特异性引物扩增CAP基因上、下游片段并从PDL2质粒上扩增筛选标记基因;利用Double-joint PCR构建基因敲除盒并通过PEG介导的原生质转化技术进行基因敲除和基因回补;以遗传霉素(G418)抗性为筛选标记并利用4对引物PCR检测获得基因敲除突变体;以潮霉素(HPH)抗性为筛选标记获得基因回补菌株;通过对菌株进行培养皿内生长试验明确基因对病菌营养生长的影响;通过离体苹果枝条接种试验分析该病菌CAP超家族基因的毒性功能。【结果】在苹果黑腐皮壳菌中鉴定到3个具有CAP保守结构域的基因,分别命名为VmPR1a、VmPR1b和VmPR1c。序列特征分析发现,3个CAP蛋白均包含4个保守区:N端信号肽、N端延伸区(NTE)、CAP保守功能域和C端延伸区(CTE)。系统发育分析显示,3个CAP蛋白聚于不同的进化支,VmPR1a聚于clade2进化支并与粗糙链孢霉(Neurospora crassa)CAP蛋白进化关系较近;VmPR1b聚于clade3且与镰孢菌属(Fusarium spp.)CAP蛋白的进化关系更近;VmPR1c聚于clade1,并且也与粗糙链孢霉的CAP蛋白进化关系较近。RT-qPCR分析结果显示,VmPR1a、VmPR1b和VmPR1c在病菌侵染早期(6 h和12 h)均显著上调表达。利用PEG遗传转化技术获得VmPR1a、VmPR1b和VmPR1c敲除突变体(ΔVmPR1a-7/23、ΔVmPR1b-20/31和ΔVmPR1c-26/40);营养生长观察发现,所有敲除突变体生长表型与野生型菌株03-8均无明显差异;致病力检测发现,ΔVmPR1b-20/31致病力较野生型无明显变化,而ΔVmPR1a-7/23和ΔVmPR1c-26/40致病力较野生型显著下降。将VmPR1a和VmPR1c分别回补至ΔVmPR1a和ΔVmPR1c,回补菌株(VmPR1a/C和VmPR1c/C)致病力恢复至野生型水平。【结论】苹果黑腐皮壳菌中存在3个CAP超家族基因(VmPR1a、VmPR1b和VmPR1c),其中VmPR1a和VmPR1c是苹果黑腐皮壳菌重要的毒性因子。

关键词：苹果黑腐皮壳菌 CAP蛋白基因鉴定基因敲除毒性功能

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双相不锈钢在不同焊接位置下接头性能研究

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金属加工（热加工） 2020年第7期 51-53页

作者：聂加俊王龙富芜湖造船厂有限公司安徽芜湖241001

采用熔化极气体保护焊对双相不锈钢2205进行焊接工艺评定试验,在横焊、立焊不同焊接位置下进行试验,通过对接头力学性能及双相组织含量测定,焊缝及热影响区的金相等数据综合分析。结果表明:适合的热输入范围可以得到良好接头性能,完全... 详细信息

采用熔化极气体保护焊对双相不锈钢2205进行焊接工艺评定试验,在横焊、立焊不同焊接位置下进行试验,通过对接头力学性能及双相组织含量测定,焊缝及热影响区的金相等数据综合分析。结果表明:适合的热输入范围可以得到良好接头性能,完全可以满足双相不锈钢焊接评定要求。

关键词：双相不锈钢焊接工艺评定双相组织热输入

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