[ Objective ] The paper was to confirrm the effect of hrpZpsg12 gene on the pathogenicity of Pseudomonas syringae pv. glycinea. [ Method ] hrpZpsg12 gene was cloned from P. syringae using PCR method. The knockout plas...[ Objective ] The paper was to confirrm the effect of hrpZpsg12 gene on the pathogenicity of Pseudomonas syringae pv. glycinea. [ Method ] hrpZpsg12 gene was cloned from P. syringae using PCR method. The knockout plasmid pKNOCK-Cm with suicide characteristics and cosmid pUFR034 with complementation func- tion were used to construct the mutation vector pKNOCK477-7 and complementary vector pUFR1026-68 of hrpZpsg12 gene, the mutant 477-1 and the functional com- plementation unit 1026-5 of the gene was also screened out. Three strains including wild-type Psg12, mutant 477-1 and complementary unit 1026-5 were simultane- ously inoculated into soybean leaves and tobacco leaves, then pathogenicity determination and hypersensitive reaction analysis were carried out. [ Result] All the inoculated leaves of soybean and tobacco produced reaction lesion. However, the sizes of reaction lesion were different. The lesion in the leaves inoculated with Psgl2 was relatively large, while the lesion in the leaves inoculated with 477-1 was relatively small; the lesion of complementary unit 1026-5 was similar to wild- type Psgl2. Analysis of reproduction quantity of bacteria in lesions showed that the reproduction quantity of wild-type Psg12 was the highest, while that of mutant 477-1 was the lowest. The reproduction quantity of complementary unit 1026-5 was similar to that of wild-type Psg12. [ Conclusion] hrpZpsg12 gene could enhance the pathogenicity of P. syrimgae on Soybean and produce hypersensitive response in tobacco.展开更多
Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis （phz） genes in two ge...Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis （phz） genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato （Pst） DC3000 and Xanthomonas oryzae pv. oryzae （Xoo） PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-l-carboxylic acid （PCA） of Pst DC3000 accumulated to 13.4 IJg L-1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL-1 PCA, while Xoo PXO99A was sensitive to 50 IJg mL ~ PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.展开更多
This study was done to determine the causal organism of the pear blossom and bud blast in China. It was identified by a bacteriological test, electro-microscopic observation, Koch＇s postulate test, Biolog, fatty acid...This study was done to determine the causal organism of the pear blossom and bud blast in China. It was identified by a bacteriological test, electro-microscopic observation, Koch＇s postulate test, Biolog, fatty acid methyl esters （FAMEs）, and a polymerase chain reaction （PCR） test, and compared with the standard reference strains. Six representative strains out of 20 pathogenic bacterial isolates from 16 diseased samples showed characteristics similar to three standard strains of Pseudomonas syringae pv. syringae from Belgium. They were identified as P. syringae pv. syringae with a Biolog similarity of 0.57-0.86 and FAMEs similarity of 0.58-0.81. The bacterium was reisolated from the symptomatic plants and blossoms. Identification as P. syringae pv. syringae was confirmed by using PCR primers and sequence tests, and compared with the above-mentioned results. The data supported the fact that the pear blossom and bud blast in China could be caused by P. syringae pv. syringae.展开更多
Soybean bacterial spot disease caused by Pseudomonas syringae pv.Glycinea which is a bacterial disease seriously affects soybean yield.Ten soybean germplasms and recombinant inbred lines(RILs)population were used to i...Soybean bacterial spot disease caused by Pseudomonas syringae pv.Glycinea which is a bacterial disease seriously affects soybean yield.Ten soybean germplasms and recombinant inbred lines(RILs)population were used to identify the resistant trait after inoculated with P.sg(P.sgneau001)in this study.High-density genetic mapping was obtained by specific length amplified fragment sequencing(SLAF-seq)of 149 RILs population which was derived from the crossing between Charleston and Dongnong594.The results indicated that 10 germplasm resources had four resistant germplasms included highly resistant cultivar Charleston,four susceptible varieties included Dongnong594 and two moderately resistant cultivars.Five quantitative trait locus(QTLs)were detected in RILs population by the composite interval mapping(CIM)method,and located on Linkage Group(LG)D1b(chromosome two),LG C2(chromosome six)and LG H(chromosome 12),respectively.LOD scores ranged from 2.68 to 4.95 and the phenotypic variation percentage was from 6%to 11%.Six candidate genes were detected,according to the result of gene annotation information.Four of them had relationship with protein kinase activity,protein phosphorylation and leucine rich repeat(LRR)transmembrane protein,which had high expression after inoculated with P.sg by qRT-PCR.展开更多
Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to es...Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.展开更多
Kiwifruit canker disease seriously affects the yield and quality of"Guichang"kiwifruit in Xiuwen County,Guizhou Province.In order to scientifically,safely,greenly and efficiently prevent and control the dise...Kiwifruit canker disease seriously affects the yield and quality of"Guichang"kiwifruit in Xiuwen County,Guizhou Province.In order to scientifically,safely,greenly and efficiently prevent and control the disease,theory was combined with prevention and control techniques to optimize existing prevention and control techniques,so as to improve the production yield and quality of kiwifruit.Specifically,biocontrol strains targeting local kiwifruit canker disease were screened,and reduced and mixed use of agrochemicals with improved efficiency was studied;and the effects and application techniques of disease resistance inducers and bioorganic fertilizers in inducing systemic disease resistance in kiwifruit trees were explored,and finally,an integrated green prevention and control scheme for kiwifruit canker disease that is suitable for kiwifruit production areas in Guizhou Province and has strong operability was proposed.This study provides technical support for green,efficient,standardized production technical services and sustainable and healthy development of kiwifruit industry.展开更多
对不同猕猴桃品种的分子生物学试验表明:猕猴桃的DNA浓度在920μg/mL符合RAPD分析的要求。通过60个随机引物的PCR扩增,报道了6个不同品种和类型猕猴桃种质资源的RAPD多态性,计算了它们之间的遗传距离,构建了聚类图,并讨论了其亲缘关系...对不同猕猴桃品种的分子生物学试验表明:猕猴桃的DNA浓度在920μg/mL符合RAPD分析的要求。通过60个随机引物的PCR扩增,报道了6个不同品种和类型猕猴桃种质资源的RAPD多态性,计算了它们之间的遗传距离,构建了聚类图,并讨论了其亲缘关系。聚类分析图反映出来源于安徽省主要猕猴桃产区的6个样品可以分为3组,其中抗病与感病的相对较为集中,由此可推断出现这种聚类的原因可能是由于它们基因组中有相同的DNA片段。抗病品系都有一条1 458 bp DNA片段,而感病品系均没有该带。故该片段可能与猕猴桃植株抗溃疡病相关。RAPD多态性从分子水平上反映出了猕猴桃种质资源不同品种及不同类型间复杂的遗传背景,为抗病育种的亲本选配提供了依据,也为合成猕猴桃抗溃疡病探针并用于检测猕猴桃抗溃疡病种质和分子标记辅助育种奠定了基础。展开更多
基金Supported by Scientific Research Foundation Project of Jilin Agricultural University" hrpZ Psg12 Protein Function of Pseudomonas syringae pv.glycinea" (384)Major Project of Cultivation of Genetically Modified Biological New Varieties of "Eleventh Five-Year Plan" of Ministry of Agriculture"Cultivation of New Transgenic Varieties of Soybean with Diseases and Pests Resistance"(2008ZX08004-004)~~
文摘[ Objective ] The paper was to confirrm the effect of hrpZpsg12 gene on the pathogenicity of Pseudomonas syringae pv. glycinea. [ Method ] hrpZpsg12 gene was cloned from P. syringae using PCR method. The knockout plasmid pKNOCK-Cm with suicide characteristics and cosmid pUFR034 with complementation func- tion were used to construct the mutation vector pKNOCK477-7 and complementary vector pUFR1026-68 of hrpZpsg12 gene, the mutant 477-1 and the functional com- plementation unit 1026-5 of the gene was also screened out. Three strains including wild-type Psg12, mutant 477-1 and complementary unit 1026-5 were simultane- ously inoculated into soybean leaves and tobacco leaves, then pathogenicity determination and hypersensitive reaction analysis were carried out. [ Result] All the inoculated leaves of soybean and tobacco produced reaction lesion. However, the sizes of reaction lesion were different. The lesion in the leaves inoculated with Psgl2 was relatively large, while the lesion in the leaves inoculated with 477-1 was relatively small; the lesion of complementary unit 1026-5 was similar to wild- type Psgl2. Analysis of reproduction quantity of bacteria in lesions showed that the reproduction quantity of wild-type Psg12 was the highest, while that of mutant 477-1 was the lowest. The reproduction quantity of complementary unit 1026-5 was similar to that of wild-type Psg12. [ Conclusion] hrpZpsg12 gene could enhance the pathogenicity of P. syrimgae on Soybean and produce hypersensitive response in tobacco.
基金supported by the grants from the Genetically Modified Organisms Breeding Major Projects, China (2014ZX0800905B)the Fundamental Research Funds for the Central Universities, Chinathe Program for New Century 151 Talents of Zhejiang Province, China
文摘Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis （phz） genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato （Pst） DC3000 and Xanthomonas oryzae pv. oryzae （Xoo） PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-l-carboxylic acid （PCA） of Pst DC3000 accumulated to 13.4 IJg L-1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL-1 PCA, while Xoo PXO99A was sensitive to 50 IJg mL ~ PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.
基金the National High Technology Research and Development Program of China (2006AA10A211)National Natural Science Foundation of China (30671397)Hangzhou Agricultural Development Foundation,China (2007-2008)
文摘This study was done to determine the causal organism of the pear blossom and bud blast in China. It was identified by a bacteriological test, electro-microscopic observation, Koch＇s postulate test, Biolog, fatty acid methyl esters （FAMEs）, and a polymerase chain reaction （PCR） test, and compared with the standard reference strains. Six representative strains out of 20 pathogenic bacterial isolates from 16 diseased samples showed characteristics similar to three standard strains of Pseudomonas syringae pv. syringae from Belgium. They were identified as P. syringae pv. syringae with a Biolog similarity of 0.57-0.86 and FAMEs similarity of 0.58-0.81. The bacterium was reisolated from the symptomatic plants and blossoms. Identification as P. syringae pv. syringae was confirmed by using PCR primers and sequence tests, and compared with the above-mentioned results. The data supported the fact that the pear blossom and bud blast in China could be caused by P. syringae pv. syringae.
基金Supported by the National Key R&D Program of China(2016YFD0100201)Science Foundation for Distinguished Young Scholars of Heilongjiang Province(JC2016004)Harbin Science Technology Project(2015RQXXJ018)。
文摘Soybean bacterial spot disease caused by Pseudomonas syringae pv.Glycinea which is a bacterial disease seriously affects soybean yield.Ten soybean germplasms and recombinant inbred lines(RILs)population were used to identify the resistant trait after inoculated with P.sg(P.sgneau001)in this study.High-density genetic mapping was obtained by specific length amplified fragment sequencing(SLAF-seq)of 149 RILs population which was derived from the crossing between Charleston and Dongnong594.The results indicated that 10 germplasm resources had four resistant germplasms included highly resistant cultivar Charleston,four susceptible varieties included Dongnong594 and two moderately resistant cultivars.Five quantitative trait locus(QTLs)were detected in RILs population by the composite interval mapping(CIM)method,and located on Linkage Group(LG)D1b(chromosome two),LG C2(chromosome six)and LG H(chromosome 12),respectively.LOD scores ranged from 2.68 to 4.95 and the phenotypic variation percentage was from 6%to 11%.Six candidate genes were detected,according to the result of gene annotation information.Four of them had relationship with protein kinase activity,protein phosphorylation and leucine rich repeat(LRR)transmembrane protein,which had high expression after inoculated with P.sg by qRT-PCR.
基金Supported by the Natural Science Foundation of Heilongjiang Province(Topic C2017032)Heilongjiang Province Applied Technology Research and Development Program(Topic GA19B104)the National Key Research and Development Program(Topic 2018YFD0300105)。
文摘Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.
基金Supported by Science and Technology Support Program of Guizhou Province(QKHZC[2020]1Y135)General Higher Education Science and Technology Top-notch Talents Project of Guizhou Province(QJH KY Z[2021]037)+5 种基金Science and Technology Program of Guizhou Province(QKHJZ-ZK[2022]ZD 025)High-level Talent Scientific Research Startup Project of Guizhou Institute of Technology(XJGC20190632)Earth Thesis Project of Guizhou Institute of Technology(KJZX20-005)High-Level Talent Initial Funding of Guizhou Industry Polytechnic College(2023-RC-01)Enterprise Commissioned Project of Guizhou Industrial Polytechnic College(2023-HX-01)Enterprise Commissioned Project of Guizhou Industrial Polytechnic College(2023-HX-02).
文摘Kiwifruit canker disease seriously affects the yield and quality of"Guichang"kiwifruit in Xiuwen County,Guizhou Province.In order to scientifically,safely,greenly and efficiently prevent and control the disease,theory was combined with prevention and control techniques to optimize existing prevention and control techniques,so as to improve the production yield and quality of kiwifruit.Specifically,biocontrol strains targeting local kiwifruit canker disease were screened,and reduced and mixed use of agrochemicals with improved efficiency was studied;and the effects and application techniques of disease resistance inducers and bioorganic fertilizers in inducing systemic disease resistance in kiwifruit trees were explored,and finally,an integrated green prevention and control scheme for kiwifruit canker disease that is suitable for kiwifruit production areas in Guizhou Province and has strong operability was proposed.This study provides technical support for green,efficient,standardized production technical services and sustainable and healthy development of kiwifruit industry.
文摘对不同猕猴桃品种的分子生物学试验表明:猕猴桃的DNA浓度在920μg/mL符合RAPD分析的要求。通过60个随机引物的PCR扩增,报道了6个不同品种和类型猕猴桃种质资源的RAPD多态性,计算了它们之间的遗传距离,构建了聚类图,并讨论了其亲缘关系。聚类分析图反映出来源于安徽省主要猕猴桃产区的6个样品可以分为3组,其中抗病与感病的相对较为集中,由此可推断出现这种聚类的原因可能是由于它们基因组中有相同的DNA片段。抗病品系都有一条1 458 bp DNA片段,而感病品系均没有该带。故该片段可能与猕猴桃植株抗溃疡病相关。RAPD多态性从分子水平上反映出了猕猴桃种质资源不同品种及不同类型间复杂的遗传背景,为抗病育种的亲本选配提供了依据,也为合成猕猴桃抗溃疡病探针并用于检测猕猴桃抗溃疡病种质和分子标记辅助育种奠定了基础。
