One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker(CBC),caused by the bacteria Xanthomonas citri subsp.citri(Xcc).Response to CBC is a complex process,with both prote...One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker(CBC),caused by the bacteria Xanthomonas citri subsp.citri(Xcc).Response to CBC is a complex process,with both proteinDNA as well as protein–protein interactions for the regulatory network.To detect such interactions in CBC resistant regulation,a citrus high-throughput screening system with 203 CBC-inducible transcription factors(TFs),were developed.Screening the upstream regulators of target by yeast-one hybrid(Y1H)methods was also performed.A regulatory module of CBC resistance was identified based on this system.One TF(CsDOF5.8)was explored due to its interactions with the 1-kb promoter fragment of CsPrx25,a resistant gene of CBC involved in reactive oxygen species(ROS)homeostasis regulation.Electrophoretic mobility shift assay(EMSA),dual-LUC assays,as well as transient overexpression of CsDOF5.8,further validated the interactions and transcriptional regulation.The CsDOF5.8–CsPrx25 promoter interaction revealed a complex pathway that governs the regulation of CBC resistance via H2O2homeostasis.The high-throughput Y1H/Y2H screening system could be an efficient tool for studying regulatory pathways or network of CBC resistance regulation.In addition,it could highlight the potential of these candidate genes as targets for efforts to breed CBC-resistant citrus varieties.展开更多
Kiwifruit canker and brown spot are significant diseases affecting kiwis,caused by Pseudomonas syringae patho-genic variations(Pseudomonas syringae pv.Actinidiae(Psa))and Corynesporapolytica(Corynespora cassiicola).At ...Kiwifruit canker and brown spot are significant diseases affecting kiwis,caused by Pseudomonas syringae patho-genic variations(Pseudomonas syringae pv.Actinidiae(Psa))and Corynesporapolytica(Corynespora cassiicola).At present,the research on canker disease and brown spot disease mainly focuses on the isolation and identification of pathogenic bacteria,drug control,resistance gene mining and functional verification.Practice has proved that breeding disease resistant varieties are an effective method to control canker disease and brown spot disease.However,most existing cultivars lack genes for canker and brown spot resistance.Wild kiwifruit resources in nat-ure exhibit extensive genetic diversity due to prolonged natural selection,containing numerous resistance genes.But,due to insufficient understanding of the resistance of most kiwifruit varieties(lines)to canker disease and brown spot disease,some high-quality resources have not been fully utilized.The incidence of canker and brown spot of 18 kiwifruit cultivars(lines)was measured by inoculating isolated branches and leaves,and their resistance to canker and brown spot was analyzed according to the length,disease index,mean diameter,and systematic clustering.The results were as follows:Among 18 different kiwifruit varieties(lines)for canker disease,there were two highly resistant materials,eight disease-resistant materials,four disease-susceptible materials,and two highly susceptible materials.Moreover,regarding brown spot disease,there were one highly resistant material,five dis-ease-resistant materials,four susceptible materials,and three highly susceptible materials.Furthermore,four resources were resistant to both diseases.The outcomes provided a theoretical basis for breeding kiwifruit against canker and brown spot.展开更多
The most damaging citrus diseases are Huanglongbing(HLB)and citrus canker,which are caused by Candidatus Liberibacter asiaticus(CaLas)and Xanthomonas citri pv.citri(Xcc),respectively.Endolysins from bacteriophages are...The most damaging citrus diseases are Huanglongbing(HLB)and citrus canker,which are caused by Candidatus Liberibacter asiaticus(CaLas)and Xanthomonas citri pv.citri(Xcc),respectively.Endolysins from bacteriophages are a possible option for disease resistance in plant breeding.Here,we report improvement of citrus resistance to HLB and citrus canker using the LasLYS1 and LasLYS2 endolysins from CaLas.LasLYS2 demonstrated bactericidal efficacy against several Rhizobiaceae bacteria and Xcc,according to inhibition zone analyses.The two genes,driven by a strong promoter from Cauliflower mosaic virus,35S,were integrated into Carrizo citrange via Agrobacterium-mediated transformation.More than 2 years of greenhouse testing indicated that LasLYS2 provided substantial and long-lasting resistance to HLB,allowing transgenic plants to retain low CaLas titers and no obvious symptoms while also clearing CaLas from infected plants in the long term.LasLYS2 transgenic plants with improved HLB resistance also showed resistance to Xcc,indicating that LasLYS2 had dual resistance to HLB and citrus canker.A microbiome study of transgenic plants revealed that the endolysins repressed Xanthomonadaceae and Rhizobiaceae populations in roots while increasing Burkholderiaceae and Rhodanobacteraceae populations,which might boost the citrus defense response,according to transcriptome analysis.We also found that Lyz domain 2 is the key bactericidal motif of LasLYS1 and LasLYS2.Four endolysins with potential resistance to HLB and citrus canker were found based on the structures of LasLYS1 and LasLYS2.Overall,the work shed light on the mechanisms of resistance of CaLas-derived endolysins,providing insights for designing endolysins to develop broad-spectrum disease resistance in citrus.展开更多
为全面系统了解林木腐烂病国内外研究现状和进展情况,检索Web of Science(WOS)核心集和中国知网(CNKI)1990—2022年收录的林木腐烂病相关研究文献,运用CiteSpace 5.8.R3软件对纳入文献的国家、机构、作者共现、关键词聚类及突现等数据...为全面系统了解林木腐烂病国内外研究现状和进展情况,检索Web of Science(WOS)核心集和中国知网(CNKI)1990—2022年收录的林木腐烂病相关研究文献,运用CiteSpace 5.8.R3软件对纳入文献的国家、机构、作者共现、关键词聚类及突现等数据进行可视化分析。经筛选,共纳入WOS文献621篇、CNKI文献1886篇。国际上林木腐烂病研究热度总体呈上升趋势,研究国家以中、美为主,研究机构以高校为主,国内机构和学者非常活跃,合作较多,国家之间合作不足。目前较活跃的高频关键词聚类为taxonomy(分类)、antifungal activity(抗真菌活性)、mycovirus(真菌病毒)和病虫害等。国际上林木腐烂病研究热点及趋势在维度上不断发展,聚焦bark(树皮)、purification(纯化)、lectin(凝集素)、diversity(多样性)等热点问题,国内研究集中于福美砷、木质部、树势、桥接等传统角度,并逐渐关注孢子角、发病规律、抑菌活性等主题的中国国情下的研究。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
新疆阿克苏地区由Valsa mali var.pyri、Valsa sordida引起的腐烂病是制约库尔勒香梨发展的重要病害,为研究库尔勒香梨感染腐烂病后与防御酶活性的变化关系,以库尔勒香梨为试验材料,分别采集同一株树不同发病组织和不同发病程度枝条韧皮...新疆阿克苏地区由Valsa mali var.pyri、Valsa sordida引起的腐烂病是制约库尔勒香梨发展的重要病害,为研究库尔勒香梨感染腐烂病后与防御酶活性的变化关系,以库尔勒香梨为试验材料,分别采集同一株树不同发病组织和不同发病程度枝条韧皮部,研究其抗性相关酶活性变化。结果表明:病害在完全侵入组织后,SOD活性呈下降的趋势,在发病组织,POD、CAT活性最高,PAL在病健交界处活性最高。随着发病程度的增加,SOD活性整体呈先上升后下降的趋势,POD活性呈逐渐上升的趋势,CAT活性先下降后上升,PAL活性先上升后下降,PPO总体呈下降趋势。库尔勒香梨抗性相关酶活性呈现不同变化响应Valsa侵染。在Valsa与库尔勒香梨互作系统中,抗性酶可作为抗病性和检测病害指标,为检测库尔勒香梨腐烂病发病程度提供理论依据。展开更多
基金funded by the National Key Research and Development Program of China(2022YFD1201600)the earmarked fund for the China Agriculture Research System(CARS-26)+1 种基金the Fundamental Research Funds for the Central Universities,China(SWU-XDJH202308)the Science and Technology Research Program of Chongqing Municipal Education Commission,China(KJQN202001418)。
文摘One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker(CBC),caused by the bacteria Xanthomonas citri subsp.citri(Xcc).Response to CBC is a complex process,with both proteinDNA as well as protein–protein interactions for the regulatory network.To detect such interactions in CBC resistant regulation,a citrus high-throughput screening system with 203 CBC-inducible transcription factors(TFs),were developed.Screening the upstream regulators of target by yeast-one hybrid(Y1H)methods was also performed.A regulatory module of CBC resistance was identified based on this system.One TF(CsDOF5.8)was explored due to its interactions with the 1-kb promoter fragment of CsPrx25,a resistant gene of CBC involved in reactive oxygen species(ROS)homeostasis regulation.Electrophoretic mobility shift assay(EMSA),dual-LUC assays,as well as transient overexpression of CsDOF5.8,further validated the interactions and transcriptional regulation.The CsDOF5.8–CsPrx25 promoter interaction revealed a complex pathway that governs the regulation of CBC resistance via H2O2homeostasis.The high-throughput Y1H/Y2H screening system could be an efficient tool for studying regulatory pathways or network of CBC resistance regulation.In addition,it could highlight the potential of these candidate genes as targets for efforts to breed CBC-resistant citrus varieties.
基金supported by the following grants:Science and Technology Support Plan of Guizhou Province:Breeding Research and Demonstration of all-Red Bud Transformation of“GH-1”Clone of“Hong yang”Kiwifruit(Guizhou Family Combination Support[2021]General 234)the National Key Research and Development Program“Quality and Efficiency Improvement Technology Integration and Demonstration of Advantageous Characteristic Industries in Guizhou Karst Mountain Area(2021YFD1100300)”Post-Subsidy FundTask 3 of National Key Research and Development Program,Green Prevention and Control Technology Integration and Demonstration of Main Diseases and Insect Pests of Kiwifruit in Shuicheng City,China(2022YFD1601710-3).
文摘Kiwifruit canker and brown spot are significant diseases affecting kiwis,caused by Pseudomonas syringae patho-genic variations(Pseudomonas syringae pv.Actinidiae(Psa))and Corynesporapolytica(Corynespora cassiicola).At present,the research on canker disease and brown spot disease mainly focuses on the isolation and identification of pathogenic bacteria,drug control,resistance gene mining and functional verification.Practice has proved that breeding disease resistant varieties are an effective method to control canker disease and brown spot disease.However,most existing cultivars lack genes for canker and brown spot resistance.Wild kiwifruit resources in nat-ure exhibit extensive genetic diversity due to prolonged natural selection,containing numerous resistance genes.But,due to insufficient understanding of the resistance of most kiwifruit varieties(lines)to canker disease and brown spot disease,some high-quality resources have not been fully utilized.The incidence of canker and brown spot of 18 kiwifruit cultivars(lines)was measured by inoculating isolated branches and leaves,and their resistance to canker and brown spot was analyzed according to the length,disease index,mean diameter,and systematic clustering.The results were as follows:Among 18 different kiwifruit varieties(lines)for canker disease,there were two highly resistant materials,eight disease-resistant materials,four disease-susceptible materials,and two highly susceptible materials.Moreover,regarding brown spot disease,there were one highly resistant material,five dis-ease-resistant materials,four susceptible materials,and three highly susceptible materials.Furthermore,four resources were resistant to both diseases.The outcomes provided a theoretical basis for breeding kiwifruit against canker and brown spot.
基金supported by the National Key Research and Development Program of China(2021YFD1400800,2022YFD1400200)the National Natural Sciences Foundation of China(31972393)+1 种基金the Fundamental Research Funds for the Central Universities(SWU-XDJH202308,XDJK2018B016)the Earmarked Fund for China Agriculture Research System(CARS-27).
文摘The most damaging citrus diseases are Huanglongbing(HLB)and citrus canker,which are caused by Candidatus Liberibacter asiaticus(CaLas)and Xanthomonas citri pv.citri(Xcc),respectively.Endolysins from bacteriophages are a possible option for disease resistance in plant breeding.Here,we report improvement of citrus resistance to HLB and citrus canker using the LasLYS1 and LasLYS2 endolysins from CaLas.LasLYS2 demonstrated bactericidal efficacy against several Rhizobiaceae bacteria and Xcc,according to inhibition zone analyses.The two genes,driven by a strong promoter from Cauliflower mosaic virus,35S,were integrated into Carrizo citrange via Agrobacterium-mediated transformation.More than 2 years of greenhouse testing indicated that LasLYS2 provided substantial and long-lasting resistance to HLB,allowing transgenic plants to retain low CaLas titers and no obvious symptoms while also clearing CaLas from infected plants in the long term.LasLYS2 transgenic plants with improved HLB resistance also showed resistance to Xcc,indicating that LasLYS2 had dual resistance to HLB and citrus canker.A microbiome study of transgenic plants revealed that the endolysins repressed Xanthomonadaceae and Rhizobiaceae populations in roots while increasing Burkholderiaceae and Rhodanobacteraceae populations,which might boost the citrus defense response,according to transcriptome analysis.We also found that Lyz domain 2 is the key bactericidal motif of LasLYS1 and LasLYS2.Four endolysins with potential resistance to HLB and citrus canker were found based on the structures of LasLYS1 and LasLYS2.Overall,the work shed light on the mechanisms of resistance of CaLas-derived endolysins,providing insights for designing endolysins to develop broad-spectrum disease resistance in citrus.
文摘为全面系统了解林木腐烂病国内外研究现状和进展情况,检索Web of Science(WOS)核心集和中国知网(CNKI)1990—2022年收录的林木腐烂病相关研究文献,运用CiteSpace 5.8.R3软件对纳入文献的国家、机构、作者共现、关键词聚类及突现等数据进行可视化分析。经筛选,共纳入WOS文献621篇、CNKI文献1886篇。国际上林木腐烂病研究热度总体呈上升趋势,研究国家以中、美为主,研究机构以高校为主,国内机构和学者非常活跃,合作较多,国家之间合作不足。目前较活跃的高频关键词聚类为taxonomy(分类)、antifungal activity(抗真菌活性)、mycovirus(真菌病毒)和病虫害等。国际上林木腐烂病研究热点及趋势在维度上不断发展,聚焦bark(树皮)、purification(纯化)、lectin(凝集素)、diversity(多样性)等热点问题,国内研究集中于福美砷、木质部、树势、桥接等传统角度,并逐渐关注孢子角、发病规律、抑菌活性等主题的中国国情下的研究。
文摘新疆阿克苏地区由Valsa mali var.pyri、Valsa sordida引起的腐烂病是制约库尔勒香梨发展的重要病害,为研究库尔勒香梨感染腐烂病后与防御酶活性的变化关系,以库尔勒香梨为试验材料,分别采集同一株树不同发病组织和不同发病程度枝条韧皮部,研究其抗性相关酶活性变化。结果表明:病害在完全侵入组织后,SOD活性呈下降的趋势,在发病组织,POD、CAT活性最高,PAL在病健交界处活性最高。随着发病程度的增加,SOD活性整体呈先上升后下降的趋势,POD活性呈逐渐上升的趋势,CAT活性先下降后上升,PAL活性先上升后下降,PPO总体呈下降趋势。库尔勒香梨抗性相关酶活性呈现不同变化响应Valsa侵染。在Valsa与库尔勒香梨互作系统中,抗性酶可作为抗病性和检测病害指标,为检测库尔勒香梨腐烂病发病程度提供理论依据。