【目的】明确甘肃省苹果树腐烂病菌(Valsa mali)致病力分化情况。【方法】采用平板培养法和离体枝条接种法对Valsa mali两变种的培养性状和致病力进行了研究。【结果】Valsa mali在PDA上有褐色类群和乳白色类群,褐色类群为Valsa mali va...【目的】明确甘肃省苹果树腐烂病菌(Valsa mali)致病力分化情况。【方法】采用平板培养法和离体枝条接种法对Valsa mali两变种的培养性状和致病力进行了研究。【结果】Valsa mali在PDA上有褐色类群和乳白色类群,褐色类群为Valsa mali var.mali菌株,乳白色类群为Valsa mali var.pyri菌株。Valsa mali var.mali不同菌株之间存在致病力分化,菌株PL-1致病力最强,第7天病斑面积为9.54 cm2,菌株TS-5致病力最弱,第7天时病斑面积为4.7 6cm2。Valsa mali var.pyri不同菌株之间也存在致病力分化,菌株WW-1致病力最强,第7天病斑面积为2.72 cm2,菌株WW-2致病力最弱,第7天病斑面积为1.40 cm2。Valsa mali在不同品种间致病力不同,在‘新红星’致病力强,在‘富士’致病力弱。Valsa mali不但侵染苹果树还可以侵染梨树和桃树,Valsa mali var.pyri菌株在梨树上的致病力强于Valsa mali var.mali。【结论】Valsa mali两变种间致病力差异显著,褐色类群菌株Valsa mali var.mali致病力强于乳白色类群菌株Valsa mali var.pyri,Valsa mali两变种内各菌株之间也存在致病力差异显著。展开更多
林木腐烂病是苹果树、梨树和杨树等林木枝干的重要真菌性病害。为了筛选出对苹果树腐烂病菌Valsa mali var.mali、梨树腐烂病菌V.mali var.pyri和杨树腐烂病菌V.sordida等3种不同寄主腐烂病菌都能有效防控的杀菌剂,本研究开展室内毒力...林木腐烂病是苹果树、梨树和杨树等林木枝干的重要真菌性病害。为了筛选出对苹果树腐烂病菌Valsa mali var.mali、梨树腐烂病菌V.mali var.pyri和杨树腐烂病菌V.sordida等3种不同寄主腐烂病菌都能有效防控的杀菌剂,本研究开展室内毒力试验比较了7种杀菌剂对3种腐烂病病原菌菌丝生长和分生孢子萌发的抑制效果,并进一步通过田间活性测定试验比较7种杀菌剂对梨树腐烂病病斑扩展和分生孢子发生的防治效果,同时测定了增效剂8.6%聚乙二醇(PEG)对7种杀菌剂的增效作用。毒力测定结果表明,苯醚甲环唑、戊唑醇、吡唑醚菌酯和丙唑·多菌灵对3种腐烂病病原菌菌丝生长和分生孢子萌发的抑制作用较强,其中EC_(50)平均值最低的是苯醚甲环唑,而戊唑醇的MIC平均值最低,在0.33 mg/L浓度下对3种腐烂病病原菌的菌丝生长和分生孢子萌发抑制率均达到100%。田间试验结果表明,45%苯醚甲环唑SC、43%戊唑醇SC和35%丙唑·多菌灵SE对梨树腐烂病病斑扩展和分生孢子萌发的防治效果突出,其中45%苯醚甲环唑SC 30.00 mg/L对病斑扩展防治效果达到82.23%,孢子萌发抑制效果达到85.96%,田间防治效果最好。10%丙硫唑SC+8.6%PEG处理组对病斑扩展防治效果提高了15.39百分点,达到73.46%,分生孢子萌发抑制率提高了23.75百分点,达到83.06%,增效作用显著。本研究为苹果树、梨树和杨树等3种寄主腐烂病的化学防控提供了科学依据。展开更多
【目的】分析新疆生产建设兵团第一师阿拉尔市库尔勒香梨园梨树腐烂病病原菌的种群分类,通过6对保守基因序列,研究库尔勒香梨腐烂病病原菌序列之间相关性及差异性。【方法】在库尔勒香梨园内采集具有典型症状的腐烂病病样,经科赫氏法则...【目的】分析新疆生产建设兵团第一师阿拉尔市库尔勒香梨园梨树腐烂病病原菌的种群分类,通过6对保守基因序列,研究库尔勒香梨腐烂病病原菌序列之间相关性及差异性。【方法】在库尔勒香梨园内采集具有典型症状的腐烂病病样,经科赫氏法则验证获取腐烂病病原菌纯培养。采用CTAB法提取腐烂病病原菌基因组DNA,通过PCR扩增ITS、β-tubulin、EF-1α、ACT、LSU、RBP2基因并测序,利用分子系统学分析不同基因片段的系统发育关系。【结果】在阿拉尔周边混交园内库尔勒香梨腐烂病菌种为Valsa mali var.pyri(V.ambiens、Valsa mali)。种内差异与地理位置有关,国内与国外梨腐烂病分离株差异较明显。EF-1α、ACT、LSU、RBP2在进化过程中更能够体现种间差异,系统发育树分支多,存在种间差异。【结论】新疆生产建设兵团第一师阿拉尔市库尔勒香梨腐烂病病原菌种群分布和种间差异性,库尔勒香梨园的腐烂病病原菌主要为Valsa mali var.pyri、Valsa sordida。新疆南疆绿洲特色果园种植模式,以杨树等为主防护林最为常见,存在着林果树木腐烂病交叉侵染的可能。展开更多
Apple valsa canker caused by the Ascomycete fungus Valsa mali is one of the most serious diseases of apple,resulting in huge economic losses in the apple-growing area of China.Previous study found that the pathogen co...Apple valsa canker caused by the Ascomycete fungus Valsa mali is one of the most serious diseases of apple,resulting in huge economic losses in the apple-growing area of China.Previous study found that the pathogen could acidify the infected tissues to make lower ambient pH(from 6.0 to 3.5)for their successfully colonization.The pH signaling transcription factor VmPacC is required for acidification of its environment and for full virulence in V.mali.It is known that the functional cooperation of proteins secreted by V.mali plays pivotal role in its successful colonization of host plants.In this study,we used tandem mass tag(TMT)labeling coupled with LC-MS/MS-based quantitative proteomics to analyze the VmPacC-mediated pH regulation in V.mali,focusing on differentially expressed proteins(DEPs).We identified 222 DEPs specific to VmPacC deletion,and 921 DEPs specific to different pH conditions(pH 6.0 and 3.4).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses indicated that these DEPs were mainly involved in pathways associated with carbon metabolism,biosynthesis of antibiotics,citrate cycle(TCA cycle),glycolysis/gluconeogenesis,glutathione metabolism,ribosomes,and pentose phosphate pathways.Additionally,we identified 119 DEPs that were shared among the VmPacC deletion mutant and different pH conditions,which were mainly related to energy metabolism pathways,providing the energy required for the hyphal growth and responses to environmental stresses.A protein-protein interaction(PPI)network analysis indicated that most of the shared proteins were mapped to an interaction network with a medium confidence score of 0.4.Notably,one uncharacterized protein(KUI69106.1),and two known proteins(heat shock protein 60(KUI73579.1),aspartate aminotransferase(KUI73864.1))located in the core of the network were highly connected(with≥38 directed edges)with the other shared DEPs.Our results suggest that VmPacC participates in the pathogen’s regulation to ambient pH through the regulation of energy metabolism pathways such as the glycolysis/gluconeogenesis pathway and TCA cycle.Finally,we proposed a sophisticated molecular regulatory network to explain pH decrease in V.mali.Our study,by providing insights into V.mali regulating pH,helps to elucidate the mechanisms of host acidification during pathogen infection.展开更多
Conserved effectors produced by phytopathogens play critical roles in plant-microbe interactions.NIS1-like proteins represent a newly identified family of effectors distributed in multiple fungal species.However,their...Conserved effectors produced by phytopathogens play critical roles in plant-microbe interactions.NIS1-like proteins represent a newly identified family of effectors distributed in multiple fungal species.However,their biological functions in a majority of pathogenic fungi remain largely elusive and require further investigation.In this study,we characterized two NIS1-like proteins VmNIS1 and VmNIS2 from Valsa mali,the causal agent of apple Valsa canker.Both of these two proteins were predicted to be secreted.Using agroinfiltration,we found that VmNIS1 induced intense cell death,whereas VmNIS2 suppressed INF1 elicitin-triggered cell death in Nicotiana benthamiana.Treatment of N.benthamiana with VmNIS1 recombinant protein produced by Escherichia coli activated a series of immune responses and enhanced plant disease resistance against Phytophthora capsici.In contrast,VmNIS2 suppressed plant immune responses and promoted P.capsici infection when transiently expressed in N.benthamiana.Both VmNIS1 and VmNIS2 were shown to be highly induced at late stage of V.mali infection.By individually knocking out of these two genes in V.mali,however,only VmNIS2 was shown to be required for pathogen virulence as well as tolerance to oxidative stress.Notably,we further showed that C-terminal extension of VmNIS1 was essential for plant recognition and VmNIS2 may escape plant detection via sequence truncation.Our 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.展开更多
目的从喜树组织中筛选产10-羟基喜树碱的内生真菌,并对其进行鉴定。方法从喜树(Camptotheca acuminateDecne)树皮中分离、纯化得到6株内生真菌,经摇瓶发酵后,采用TLC法、HPLC和MS分析菌丝的提取物;用插片法进行形态学观察,采用分子生物...目的从喜树组织中筛选产10-羟基喜树碱的内生真菌,并对其进行鉴定。方法从喜树(Camptotheca acuminateDecne)树皮中分离、纯化得到6株内生真菌,经摇瓶发酵后,采用TLC法、HPLC和MS分析菌丝的提取物;用插片法进行形态学观察,采用分子生物学方法对XK002菌株rDNA的ITS基因(ITS-5.8S rDNA)进行PCR扩增、测序;利用BLAST软件与GenBank数据库进行相似性分析,并用Clustal X1.81软件法构建系统发育树。结果内生真菌XK002可产10-羟基喜树碱;菌株XK002在PDA培养基上生长迅速,白色绒毛状,4d能长满直径为9cm培养皿,10d后开始形成黑色的子座,产生大量的分生孢子;ITS序列与Valsa mali Miyabe et Yamada相应序列的相似性为100%,基因库接受号(Accession No.)为FJ418170。结论根据形态学和分子生物学方法鉴定可知,XK002菌株为Valsa mali Miyabe et Yamada。展开更多
通过菌丝生长速率法检测了吡唑醚菌酯、辛菌胺、甲基硫菌灵3种药剂对山西省苹果树腐烂病优势种群Valsa mali var．mali的室内毒力，并应用整树喷淋法连续2a测定了4种常用药剂对苹果树腐烂病的田间防效。室内毒力测定结果表明，吡唑醚菌...通过菌丝生长速率法检测了吡唑醚菌酯、辛菌胺、甲基硫菌灵3种药剂对山西省苹果树腐烂病优势种群Valsa mali var．mali的室内毒力，并应用整树喷淋法连续2a测定了4种常用药剂对苹果树腐烂病的田间防效。室内毒力测定结果表明，吡唑醚菌酯对苹果树腐烂病菌生长的抑制作用最佳，甲基硫菌灵次之，辛菌胺抑制作用最低，3种药剂的Ec。平均值均低于5．0μg／mL；田间防效测定结果表明，3％辛菌胺醋酸盐水剂30-40倍液和25％吡唑醚菌酯乳油1000～1500倍液的2个高浓度处理对苹果树腐烂病病疤复发和新病疤产生的防效均在75％以上，可作为喷淋法防治苹果树腐烂病的选择药剂及适用浓度。展开更多
文摘【目的】明确甘肃省苹果树腐烂病菌(Valsa mali)致病力分化情况。【方法】采用平板培养法和离体枝条接种法对Valsa mali两变种的培养性状和致病力进行了研究。【结果】Valsa mali在PDA上有褐色类群和乳白色类群,褐色类群为Valsa mali var.mali菌株,乳白色类群为Valsa mali var.pyri菌株。Valsa mali var.mali不同菌株之间存在致病力分化,菌株PL-1致病力最强,第7天病斑面积为9.54 cm2,菌株TS-5致病力最弱,第7天时病斑面积为4.7 6cm2。Valsa mali var.pyri不同菌株之间也存在致病力分化,菌株WW-1致病力最强,第7天病斑面积为2.72 cm2,菌株WW-2致病力最弱,第7天病斑面积为1.40 cm2。Valsa mali在不同品种间致病力不同,在‘新红星’致病力强,在‘富士’致病力弱。Valsa mali不但侵染苹果树还可以侵染梨树和桃树,Valsa mali var.pyri菌株在梨树上的致病力强于Valsa mali var.mali。【结论】Valsa mali两变种间致病力差异显著,褐色类群菌株Valsa mali var.mali致病力强于乳白色类群菌株Valsa mali var.pyri,Valsa mali两变种内各菌株之间也存在致病力差异显著。
文摘【目的】分析新疆生产建设兵团第一师阿拉尔市库尔勒香梨园梨树腐烂病病原菌的种群分类,通过6对保守基因序列,研究库尔勒香梨腐烂病病原菌序列之间相关性及差异性。【方法】在库尔勒香梨园内采集具有典型症状的腐烂病病样,经科赫氏法则验证获取腐烂病病原菌纯培养。采用CTAB法提取腐烂病病原菌基因组DNA,通过PCR扩增ITS、β-tubulin、EF-1α、ACT、LSU、RBP2基因并测序,利用分子系统学分析不同基因片段的系统发育关系。【结果】在阿拉尔周边混交园内库尔勒香梨腐烂病菌种为Valsa mali var.pyri(V.ambiens、Valsa mali)。种内差异与地理位置有关,国内与国外梨腐烂病分离株差异较明显。EF-1α、ACT、LSU、RBP2在进化过程中更能够体现种间差异,系统发育树分支多,存在种间差异。【结论】新疆生产建设兵团第一师阿拉尔市库尔勒香梨腐烂病病原菌种群分布和种间差异性,库尔勒香梨园的腐烂病病原菌主要为Valsa mali var.pyri、Valsa sordida。新疆南疆绿洲特色果园种植模式,以杨树等为主防护林最为常见,存在着林果树木腐烂病交叉侵染的可能。
基金supported by grants from the National Natural Science Foundation of China(Grant No.31772115 and U1903206).
文摘Apple valsa canker caused by the Ascomycete fungus Valsa mali is one of the most serious diseases of apple,resulting in huge economic losses in the apple-growing area of China.Previous study found that the pathogen could acidify the infected tissues to make lower ambient pH(from 6.0 to 3.5)for their successfully colonization.The pH signaling transcription factor VmPacC is required for acidification of its environment and for full virulence in V.mali.It is known that the functional cooperation of proteins secreted by V.mali plays pivotal role in its successful colonization of host plants.In this study,we used tandem mass tag(TMT)labeling coupled with LC-MS/MS-based quantitative proteomics to analyze the VmPacC-mediated pH regulation in V.mali,focusing on differentially expressed proteins(DEPs).We identified 222 DEPs specific to VmPacC deletion,and 921 DEPs specific to different pH conditions(pH 6.0 and 3.4).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses indicated that these DEPs were mainly involved in pathways associated with carbon metabolism,biosynthesis of antibiotics,citrate cycle(TCA cycle),glycolysis/gluconeogenesis,glutathione metabolism,ribosomes,and pentose phosphate pathways.Additionally,we identified 119 DEPs that were shared among the VmPacC deletion mutant and different pH conditions,which were mainly related to energy metabolism pathways,providing the energy required for the hyphal growth and responses to environmental stresses.A protein-protein interaction(PPI)network analysis indicated that most of the shared proteins were mapped to an interaction network with a medium confidence score of 0.4.Notably,one uncharacterized protein(KUI69106.1),and two known proteins(heat shock protein 60(KUI73579.1),aspartate aminotransferase(KUI73864.1))located in the core of the network were highly connected(with≥38 directed edges)with the other shared DEPs.Our results suggest that VmPacC participates in the pathogen’s regulation to ambient pH through the regulation of energy metabolism pathways such as the glycolysis/gluconeogenesis pathway and TCA cycle.Finally,we proposed a sophisticated molecular regulatory network to explain pH decrease in V.mali.Our study,by providing insights into V.mali regulating pH,helps to elucidate the mechanisms of host acidification during pathogen infection.
基金supported by China Postdoctoral Science Foundation(2021 M690128)the Open Project Program of State Key Laboratory of Crop Stress Biology for Arid Areas(CSBAA2020011)+1 种基金National Natural Science Foundation-Xinjiang Joint Foundation of China(U1903206)Major Scientific and Technological Projects of Shaanxi Province(2020zdzx03-03-01).
文摘Conserved effectors produced by phytopathogens play critical roles in plant-microbe interactions.NIS1-like proteins represent a newly identified family of effectors distributed in multiple fungal species.However,their biological functions in a majority of pathogenic fungi remain largely elusive and require further investigation.In this study,we characterized two NIS1-like proteins VmNIS1 and VmNIS2 from Valsa mali,the causal agent of apple Valsa canker.Both of these two proteins were predicted to be secreted.Using agroinfiltration,we found that VmNIS1 induced intense cell death,whereas VmNIS2 suppressed INF1 elicitin-triggered cell death in Nicotiana benthamiana.Treatment of N.benthamiana with VmNIS1 recombinant protein produced by Escherichia coli activated a series of immune responses and enhanced plant disease resistance against Phytophthora capsici.In contrast,VmNIS2 suppressed plant immune responses and promoted P.capsici infection when transiently expressed in N.benthamiana.Both VmNIS1 and VmNIS2 were shown to be highly induced at late stage of V.mali infection.By individually knocking out of these two genes in V.mali,however,only VmNIS2 was shown to be required for pathogen virulence as well as tolerance to oxidative stress.Notably,we further showed that C-terminal extension of VmNIS1 was essential for plant recognition and VmNIS2 may escape plant detection via sequence truncation.Our 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.
文摘目的从喜树组织中筛选产10-羟基喜树碱的内生真菌,并对其进行鉴定。方法从喜树(Camptotheca acuminateDecne)树皮中分离、纯化得到6株内生真菌,经摇瓶发酵后,采用TLC法、HPLC和MS分析菌丝的提取物;用插片法进行形态学观察,采用分子生物学方法对XK002菌株rDNA的ITS基因(ITS-5.8S rDNA)进行PCR扩增、测序;利用BLAST软件与GenBank数据库进行相似性分析,并用Clustal X1.81软件法构建系统发育树。结果内生真菌XK002可产10-羟基喜树碱;菌株XK002在PDA培养基上生长迅速,白色绒毛状,4d能长满直径为9cm培养皿,10d后开始形成黑色的子座,产生大量的分生孢子;ITS序列与Valsa mali Miyabe et Yamada相应序列的相似性为100%,基因库接受号(Accession No.)为FJ418170。结论根据形态学和分子生物学方法鉴定可知,XK002菌株为Valsa mali Miyabe et Yamada。
文摘通过菌丝生长速率法检测了吡唑醚菌酯、辛菌胺、甲基硫菌灵3种药剂对山西省苹果树腐烂病优势种群Valsa mali var．mali的室内毒力，并应用整树喷淋法连续2a测定了4种常用药剂对苹果树腐烂病的田间防效。室内毒力测定结果表明，吡唑醚菌酯对苹果树腐烂病菌生长的抑制作用最佳，甲基硫菌灵次之，辛菌胺抑制作用最低，3种药剂的Ec。平均值均低于5．0μg／mL；田间防效测定结果表明，3％辛菌胺醋酸盐水剂30-40倍液和25％吡唑醚菌酯乳油1000～1500倍液的2个高浓度处理对苹果树腐烂病病疤复发和新病疤产生的防效均在75％以上，可作为喷淋法防治苹果树腐烂病的选择药剂及适用浓度。
