[ 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
Pseudomonas syringae pv.actinidiae(Psa)causes bacterial canker,a devastating disease threatening the Actinidia fruit industry.In a search for non-host resistance genes against Psa,we find that the nucleotidebinding le...Pseudomonas syringae pv.actinidiae(Psa)causes bacterial canker,a devastating disease threatening the Actinidia fruit industry.In a search for non-host resistance genes against Psa,we find that the nucleotidebinding leucine-rich repeat receptor(NLR)protein ZAR1 from both Arabidopsis and Nicotiana benthamiana(Nb)recognizes Hop Z5 and triggers cell death.The recognition requires ZED1 in Arabidopsis and JIM2 in Nb plants,which are members of the ZRK pseudokinases and known components of the ZAR1 resistosome.Surprisingly,Arabidopsis ZAR1 and RPM1,another NLR known to recognize Hop Z5,confer disease resistance to Hop Z5 in a strain-specific manner.Thus,ZAR1,but not RPM1,is solely required for resistance to P.s.maculicola ES4326(Psm)carrying hop Z5,whereas RPM1 is primarily required for resistance to P.s.tomato DC3000(Pst)carrying hop Z5.Furthermore,the ZAR1-mediated resistance to Psm hop Z5 in Arabidopsis is insensitive to SOBER1,which encodes a deacetylase known to suppress the RPM1-mediated resistance to Pst hop Z5.In addition,hop Z5 enhances P.syringae virulence in the absence of ZAR1 or RPM1 and that SOBER1 abolishes such virulence function.Together the study suggests that ZAR1 may be used for improving Psa resistance in Actinidia and uncovers previously unknown complexity of effectortriggered immunity and effector-triggered virulence.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
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
The aim of the study was to investigate the hepatoprotective effects of Folium syringae(FS) extracts against ethanolinduced acute liver injury. Mice and primary hepatocytes were pretreated with FS extracts at differen...The aim of the study was to investigate the hepatoprotective effects of Folium syringae(FS) extracts against ethanolinduced acute liver injury. Mice and primary hepatocytes were pretreated with FS extracts at different dosages before ethanol administration. Transaminases, glutathione S-transferase A1 level and hepatic biochemical indices(malondialdehyde, superoxide dismutase, glutathione and glutathione peroxidase) were determined. Pretreatment with FS extracts significantly inhibited the damage caused by ethanol and the hepatoprotective effects of FS were almost similar to Silymarin that was used to treat alcoholic liver injury. GSTA1 contents in all the FS extract-treated groups were significantly different from those in the ethanol-induced acute liver injury model group(p<0.01), and similar trends were observed in transaminases and hepatic indices level both in vitro and in vivo. The results showed that FS extracts had hepatoprotective effects against ethanol-induced injury. Those effects might be related to the enhancement of antioxidant capacity of liver cells, and FS extracts could reduce the release of liver GSTA1, which contributed to improve liver detoxification.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
Mulberry bacterial blight is caused by Pseudomonas syringae pv. mori. Coronatine （COR）, a phytotoxin and phytohormone produced by several strains of Pseudomonas syringae, is suggested to have an important role in pa...Mulberry bacterial blight is caused by Pseudomonas syringae pv. mori. Coronatine （COR）, a phytotoxin and phytohormone produced by several strains of Pseudomonas syringae, is suggested to have an important role in pathogen-plant interaction. The aim of our study was to examine the influence of COR on mul- berry in the process of pathogen infection. Results showed that COR could suppress stomatal closure induced by pathogen-associated molecular patterns （PAMPS）, assist pathogenic bacteria into the leaves, and promote bacterial proliferation in the tissues. High-concentration （1 μmol/L） exogenous COR and COR-producing bacterial pathogen induced chlorosis symptom and decrease of chlorophyll content, contrary to the effects of low-concentration （0.001 μmol/L） exogenous COR and non-COR-preducing bacterial pathogen. Treatments with COR and DC3000 enhanced the production of reactive oxygen species （ ROS）, namely, hydrogen peroxide （H2O2 ） and superexide anion （O2-）, but there were two H2O2 peaks at 1 -3 hpi and 8 -24 hpi in the DC3000-treated leaves while only one peak at 1 -3 h was observed 1 -3 h in the COR-treated ones. H202 could kill the pathogenic bacteria, on the other hand, it also acted as an upstream signaling molecule to promote nitric oxide （NO） production to further participate in the signaling pathway. Enzymatic antioxidant systems （superoxide dismutase, peroxidase and catalase） and defensive enzyme systems （lipoxygenase, polyphenol oxidase and phenylalnine ammonialyase） were activated by COR. Therefore, COR could cooperate on the inva- sion and proliferation of COR-producing bacterial pathogens, and induce the chloresis symptom in mulberry. At the same time, exogenously applied COR also could enhance the resistance to P. syringae pv. mori by production of signal molecules to activate signaling pathway and promoting defense-related metabolism.展开更多
Some Pseudomonas syringae pathovars secrete tabtoxin, a monocyclic β-lactam antibiotic, responsible for chlorosis, the principal halo blight symptom in susceptible plants as oats, rye, barley, wheat and sorghum, amon...Some Pseudomonas syringae pathovars secrete tabtoxin, a monocyclic β-lactam antibiotic, responsible for chlorosis, the principal halo blight symptom in susceptible plants as oats, rye, barley, wheat and sorghum, among other. Here, we demonstrated that the production of tabtoxin in a P. syringae strain increased at least 150%, when choline, betaine or dimethylglycine were used as nitrogen source, or when choline was added as osmoprotectant in hyperosmolar culture media. Besides, we investigated the induction of phosphorylcholine phosphatase (PchP) activity when choline or its metabolites were used as nitrogen sources. PchP is an enzyme involved in Pseudomonas aeruginosa pathogenesis through its contribution to the breakdown of choline-containing compounds of the host cells. Considering these results and that the success of a pathogenic microorganism depends on its ability to survive and proliferate in its target tissue, we propose that choline is one of the plant signals that contribute to establishment of the infection by tabtoxin-producing strains of P. syringae.展开更多
Pseudomonas syringae pv.actinidiae(Psa)causes destructive kiwifruit bacterial canker by invading vascular tissues across multiple plant organs.However,the cellular mechanism underlying its systemic transmission and ce...Pseudomonas syringae pv.actinidiae(Psa)causes destructive kiwifruit bacterial canker by invading vascular tissues across multiple plant organs.However,the cellular mechanism underlying its systemic transmission and cell-to-cell movement within these specialized vascular conduits remains unclear.In this study,a Psa-GFP strain and various microscopic techniques were used to investigate the interaction between kiwifruit and Psa.Our results reveal that Psa strategically exploits host vascular conduits for systemic movement,with the xylem vessel being the predominant avenue.In the phloem,Psa exhibits adaptive alteration in bacterial shape to traverse sieve pores,facilitating its systemic spread along sieve tubes and inducing phloem necrosis.Within the xylem,Psa breaches pit membranes to migrate between adjacent vessels.Furthermore,phloem fibers act as an initial barrier at the early stages of infection,delaying Psa's entry into vascular tissues during its journey to the xylem.Additionally,at the junctions of stemestem or stem-leaf,branch trace or leaf trace mediates the bacterial organ-to-organ translocation,thus facilitating the systemic progression of disease.In conclusion,our findings shed light on the cellular mechanism employed by Psa to exploit the woody plant's vascular network for infection,thereby enhancing a better understanding of the biology of this poorly defined bacterium.These insights carry implications for the pathogenesis of Psa and other vascular pathogens,offering theoretical guidance for effective control strategies.展开更多
Many Gram-negative pathogens employ the type III secretion system(T3SS)to deliver effector proteins into host cells,thereby modulating host cellular processes and suppressing host immunity to facilitate pathogenesis a...Many Gram-negative pathogens employ the type III secretion system(T3SS)to deliver effector proteins into host cells,thereby modulating host cellular processes and suppressing host immunity to facilitate pathogenesis and colonization.In this study,we developed a straightforward,rapid,and quantitative method for detecting T3SS-mediated translocation of Pseudomonas syringae effectors using a self-assembling split Nano luciferase(Nluc)-based reporter system.It was demonstrated that this system can detect effector secretion in vitro with an exceptionally high signal-to-noise ratio and sensitivity,attributed to the strong affinity between the split domains of Nluc and the intense luminescence generated by functional Nluc.During natural infections,effectors fused to a small C-terminal fragment of Nluc were successfully translocated into plant cells and retained their virulence functions.Furthermore,upon infection of plants expressing the N-terminal fragment of Nluc with these P.syringae strains,functional Nluc proteins were spontaneously assembled and produced bright luminescence,demonstrating that this system enables the straightforward and rapid assessment of P.syringae T3SS-mediated effector translocation during natural infections.In conclusion,the self-assembling split Nluc-based reporting system developed in this study is suitable for efficient in vitro and in planta detection of effectors secreted via T3SS.展开更多
Ambient temperature affects the occurrence and prevalence of plant disease.Most bacterial diseases are damaging at high temperatures.However,kiwifruit bacterial canker caused by Pseudomonas syringae pv.actinidiae(Psa)...Ambient temperature affects the occurrence and prevalence of plant disease.Most bacterial diseases are damaging at high temperatures.However,kiwifruit bacterial canker caused by Pseudomonas syringae pv.actinidiae(Psa)has been found to be prevalent at relatively cool temperatures,and it is unclear how ambient temperature affects the development of kiwifruit bacterial canker.In this study,basal resistance to Psa was suppressed in kiwifruit at cool growth temperature(16℃)compared with at normal temperature(24℃).In addition,RNA sequence analysis and ethylene content assessment indicated that ethylene modulated kiwifruit resistance to Psa at normal growth temperature and that cool temperature inhibited ethylene accumulation and Psa-induced activation of the ethylene signaling pathway in kiwifruit.Virusmediated silencing of the kiwifruit ethylene signaling gene AcEIN2 suppressed kiwifruit resistance to Psa at normal growth temperature.Exogenous application of ethylene inhibitor 1-methylcyclopropene eliminated the difference in kiwifruit resistance to Psa at 16 and 24℃.Exogenous application of ethylene analogues ethephon induced resistance to Psa in kiwifruit.In conclusion,cool temperatures impair basal resistance to Psa by reducing the activation of ethylene biosynthesis and signaling in kiwifruit.The results provide clues for new strategies to control plant diseases in a context of global environmental change.展开更多
The Pto protein kinase from Solanum pimpinellifolium interacts with Pseudomonas syringae effectors AvrPto or AvrPtoB to activate effector-triggered immunity. The previously solved crystal structures of the AvrPto-Pto ...The Pto protein kinase from Solanum pimpinellifolium interacts with Pseudomonas syringae effectors AvrPto or AvrPtoB to activate effector-triggered immunity. The previously solved crystal structures of the AvrPto-Pto and AvrPtoB-Pto complexes revealed that Pro binds each effector through both a shared and a unique interface. Hera we use natural variation in wild species of tomato to further investigate Pto recognition of these two effectors. One species, Solanum chmielewskU, was found to have many accessions that recognize only AvrPtoB. The Pto ortholog from one of these accessions was responsible for recognition of AvrPtoB and it differed from Solanum pimpinellifolium Pto by only 14 amino acids, including two in the AvrPto-specific interface, glutamate-49/glycine-51. Converting these two residues to those in Pro （histidine-49/valine-51） did not restore recognition of AvrPto. Subsequent experiments revealed that a single substitution of a histidine-to-aspartate at position 193 in Pto, which is not near the AvrPto- specific interface, was sufficient for conferring recognRion of AvrPto in plant cells. The reciprocal substi- tution of aspartate-to-histidine-193 in Pto abolished AvrPto recognition, confirming the importance of this residue. Our results reveal new aspects about effector recognition by Pto and demonstrate the value of using natural variation to understand the interaction between resistance proteins and pathogen effectors.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 grants from the National Key R&D Program of China (2021YFA1300701) to J.M.Z.the National Natural Science Foundation of China (31872654) to Z.Y.Z.the Hainan Excellent Talent Team, and the State Key Laboratory of Plant Genomics (SKLPG2016B-2) to J.M.Z
文摘Pseudomonas syringae pv.actinidiae(Psa)causes bacterial canker,a devastating disease threatening the Actinidia fruit industry.In a search for non-host resistance genes against Psa,we find that the nucleotidebinding leucine-rich repeat receptor(NLR)protein ZAR1 from both Arabidopsis and Nicotiana benthamiana(Nb)recognizes Hop Z5 and triggers cell death.The recognition requires ZED1 in Arabidopsis and JIM2 in Nb plants,which are members of the ZRK pseudokinases and known components of the ZAR1 resistosome.Surprisingly,Arabidopsis ZAR1 and RPM1,another NLR known to recognize Hop Z5,confer disease resistance to Hop Z5 in a strain-specific manner.Thus,ZAR1,but not RPM1,is solely required for resistance to P.s.maculicola ES4326(Psm)carrying hop Z5,whereas RPM1 is primarily required for resistance to P.s.tomato DC3000(Pst)carrying hop Z5.Furthermore,the ZAR1-mediated resistance to Psm hop Z5 in Arabidopsis is insensitive to SOBER1,which encodes a deacetylase known to suppress the RPM1-mediated resistance to Pst hop Z5.In addition,hop Z5 enhances P.syringae virulence in the absence of ZAR1 or RPM1 and that SOBER1 abolishes such virulence function.Together the study suggests that ZAR1 may be used for improving Psa resistance in Actinidia and uncovers previously unknown complexity of effectortriggered immunity and effector-triggered virulence.
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 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Supported by the National Natural Science Foundation of China(31472241)the Application Technology Research and Development Projects in Heilongjiang Province of China(PC13S03)
文摘The aim of the study was to investigate the hepatoprotective effects of Folium syringae(FS) extracts against ethanolinduced acute liver injury. Mice and primary hepatocytes were pretreated with FS extracts at different dosages before ethanol administration. Transaminases, glutathione S-transferase A1 level and hepatic biochemical indices(malondialdehyde, superoxide dismutase, glutathione and glutathione peroxidase) were determined. Pretreatment with FS extracts significantly inhibited the damage caused by ethanol and the hepatoprotective effects of FS were almost similar to Silymarin that was used to treat alcoholic liver injury. GSTA1 contents in all the FS extract-treated groups were significantly different from those in the ethanol-induced acute liver injury model group(p<0.01), and similar trends were observed in transaminases and hepatic indices level both in vitro and in vivo. The results showed that FS extracts had hepatoprotective effects against ethanol-induced injury. Those effects might be related to the enhancement of antioxidant capacity of liver cells, and FS extracts could reduce the release of liver GSTA1, which contributed to improve liver detoxification.
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 Science and Technology Support Program of the Jiangsu Province(BE2012365)the Modern Agro-industry Technology Research System of China(CARS-22)
文摘Mulberry bacterial blight is caused by Pseudomonas syringae pv. mori. Coronatine （COR）, a phytotoxin and phytohormone produced by several strains of Pseudomonas syringae, is suggested to have an important role in pathogen-plant interaction. The aim of our study was to examine the influence of COR on mul- berry in the process of pathogen infection. Results showed that COR could suppress stomatal closure induced by pathogen-associated molecular patterns （PAMPS）, assist pathogenic bacteria into the leaves, and promote bacterial proliferation in the tissues. High-concentration （1 μmol/L） exogenous COR and COR-producing bacterial pathogen induced chlorosis symptom and decrease of chlorophyll content, contrary to the effects of low-concentration （0.001 μmol/L） exogenous COR and non-COR-preducing bacterial pathogen. Treatments with COR and DC3000 enhanced the production of reactive oxygen species （ ROS）, namely, hydrogen peroxide （H2O2 ） and superexide anion （O2-）, but there were two H2O2 peaks at 1 -3 hpi and 8 -24 hpi in the DC3000-treated leaves while only one peak at 1 -3 h was observed 1 -3 h in the COR-treated ones. H202 could kill the pathogenic bacteria, on the other hand, it also acted as an upstream signaling molecule to promote nitric oxide （NO） production to further participate in the signaling pathway. Enzymatic antioxidant systems （superoxide dismutase, peroxidase and catalase） and defensive enzyme systems （lipoxygenase, polyphenol oxidase and phenylalnine ammonialyase） were activated by COR. Therefore, COR could cooperate on the inva- sion and proliferation of COR-producing bacterial pathogens, and induce the chloresis symptom in mulberry. At the same time, exogenously applied COR also could enhance the resistance to P. syringae pv. mori by production of signal molecules to activate signaling pathway and promoting defense-related metabolism.
文摘Some Pseudomonas syringae pathovars secrete tabtoxin, a monocyclic β-lactam antibiotic, responsible for chlorosis, the principal halo blight symptom in susceptible plants as oats, rye, barley, wheat and sorghum, among other. Here, we demonstrated that the production of tabtoxin in a P. syringae strain increased at least 150%, when choline, betaine or dimethylglycine were used as nitrogen source, or when choline was added as osmoprotectant in hyperosmolar culture media. Besides, we investigated the induction of phosphorylcholine phosphatase (PchP) activity when choline or its metabolites were used as nitrogen sources. PchP is an enzyme involved in Pseudomonas aeruginosa pathogenesis through its contribution to the breakdown of choline-containing compounds of the host cells. Considering these results and that the success of a pathogenic microorganism depends on its ability to survive and proliferate in its target tissue, we propose that choline is one of the plant signals that contribute to establishment of the infection by tabtoxin-producing strains of P. syringae.
基金supported by grants from the National Key Research and Development Program of China(Grant No.2022YFD1400200)the Special Support Plan for High-Level Talent of Shaanxi Provincethe First-Class Universities and Academic Programs of Northwest A&F University.
文摘Pseudomonas syringae pv.actinidiae(Psa)causes destructive kiwifruit bacterial canker by invading vascular tissues across multiple plant organs.However,the cellular mechanism underlying its systemic transmission and cell-to-cell movement within these specialized vascular conduits remains unclear.In this study,a Psa-GFP strain and various microscopic techniques were used to investigate the interaction between kiwifruit and Psa.Our results reveal that Psa strategically exploits host vascular conduits for systemic movement,with the xylem vessel being the predominant avenue.In the phloem,Psa exhibits adaptive alteration in bacterial shape to traverse sieve pores,facilitating its systemic spread along sieve tubes and inducing phloem necrosis.Within the xylem,Psa breaches pit membranes to migrate between adjacent vessels.Furthermore,phloem fibers act as an initial barrier at the early stages of infection,delaying Psa's entry into vascular tissues during its journey to the xylem.Additionally,at the junctions of stemestem or stem-leaf,branch trace or leaf trace mediates the bacterial organ-to-organ translocation,thus facilitating the systemic progression of disease.In conclusion,our findings shed light on the cellular mechanism employed by Psa to exploit the woody plant's vascular network for infection,thereby enhancing a better understanding of the biology of this poorly defined bacterium.These insights carry implications for the pathogenesis of Psa and other vascular pathogens,offering theoretical guidance for effective control strategies.
基金supported by grants from the Natural Science Foundation of China(32370322)the Hainan Excellent Talent Team,and the State Key Laboratory of Plant Genomics.
文摘Many Gram-negative pathogens employ the type III secretion system(T3SS)to deliver effector proteins into host cells,thereby modulating host cellular processes and suppressing host immunity to facilitate pathogenesis and colonization.In this study,we developed a straightforward,rapid,and quantitative method for detecting T3SS-mediated translocation of Pseudomonas syringae effectors using a self-assembling split Nano luciferase(Nluc)-based reporter system.It was demonstrated that this system can detect effector secretion in vitro with an exceptionally high signal-to-noise ratio and sensitivity,attributed to the strong affinity between the split domains of Nluc and the intense luminescence generated by functional Nluc.During natural infections,effectors fused to a small C-terminal fragment of Nluc were successfully translocated into plant cells and retained their virulence functions.Furthermore,upon infection of plants expressing the N-terminal fragment of Nluc with these P.syringae strains,functional Nluc proteins were spontaneously assembled and produced bright luminescence,demonstrating that this system enables the straightforward and rapid assessment of P.syringae T3SS-mediated effector translocation during natural infections.In conclusion,the self-assembling split Nluc-based reporting system developed in this study is suitable for efficient in vitro and in planta detection of effectors secreted via T3SS.
基金supported by grants from the National Key Research and Development Program of China(Grant No.2022YFD1400200)the Special Support Plan for High-Level Talent of Shaanxi Province。
文摘Ambient temperature affects the occurrence and prevalence of plant disease.Most bacterial diseases are damaging at high temperatures.However,kiwifruit bacterial canker caused by Pseudomonas syringae pv.actinidiae(Psa)has been found to be prevalent at relatively cool temperatures,and it is unclear how ambient temperature affects the development of kiwifruit bacterial canker.In this study,basal resistance to Psa was suppressed in kiwifruit at cool growth temperature(16℃)compared with at normal temperature(24℃).In addition,RNA sequence analysis and ethylene content assessment indicated that ethylene modulated kiwifruit resistance to Psa at normal growth temperature and that cool temperature inhibited ethylene accumulation and Psa-induced activation of the ethylene signaling pathway in kiwifruit.Virusmediated silencing of the kiwifruit ethylene signaling gene AcEIN2 suppressed kiwifruit resistance to Psa at normal growth temperature.Exogenous application of ethylene inhibitor 1-methylcyclopropene eliminated the difference in kiwifruit resistance to Psa at 16 and 24℃.Exogenous application of ethylene analogues ethephon induced resistance to Psa in kiwifruit.In conclusion,cool temperatures impair basal resistance to Psa by reducing the activation of ethylene biosynthesis and signaling in kiwifruit.The results provide clues for new strategies to control plant diseases in a context of global environmental change.
基金This research was supported, in part, by National Science Foundation grant IOS-1025642 （G,B.M.）,
文摘The Pto protein kinase from Solanum pimpinellifolium interacts with Pseudomonas syringae effectors AvrPto or AvrPtoB to activate effector-triggered immunity. The previously solved crystal structures of the AvrPto-Pto and AvrPtoB-Pto complexes revealed that Pro binds each effector through both a shared and a unique interface. Hera we use natural variation in wild species of tomato to further investigate Pto recognition of these two effectors. One species, Solanum chmielewskU, was found to have many accessions that recognize only AvrPtoB. The Pto ortholog from one of these accessions was responsible for recognition of AvrPtoB and it differed from Solanum pimpinellifolium Pto by only 14 amino acids, including two in the AvrPto-specific interface, glutamate-49/glycine-51. Converting these two residues to those in Pro （histidine-49/valine-51） did not restore recognition of AvrPto. Subsequent experiments revealed that a single substitution of a histidine-to-aspartate at position 193 in Pto, which is not near the AvrPto- specific interface, was sufficient for conferring recognRion of AvrPto in plant cells. The reciprocal substi- tution of aspartate-to-histidine-193 in Pto abolished AvrPto recognition, confirming the importance of this residue. Our results reveal new aspects about effector recognition by Pto and demonstrate the value of using natural variation to understand the interaction between resistance proteins and pathogen effectors.
