Lesion mmic mutants(LMMs)are plants that spontaneously form lesions without pathogeninfection or external stimulus and exhibit resistance to pathogens.Here,a rice LMM was created by ethylmethane sulfonate mutagenesis,...Lesion mmic mutants(LMMs)are plants that spontaneously form lesions without pathogeninfection or external stimulus and exhibit resistance to pathogens.Here,a rice LMM was created by ethylmethane sulfonate mutagenesis,named as hpil(hydrogen peroxide induced lesion).Diaminobenzidineand trypan blue staining showed that large amounts of H_(2)O_(2) were produced and cell death was occurredat and around the parts of lesion mimic in the rice leaves.The phenotype of hpil is controlled by a singlerecessive gene,localized at a 2 Mb interval on chromosome 2.The data suggested that hpil is a novelLMM with enhanced bacterial and fungal disease resistance,and multiple pathogenesis-related proteins(PRs)were up-regulated.The proteomes of leaves at three positions(different degrees of lesion mimicseverity)were characterized in hpil compared with its wild type plant.Differentially expressed proteinswere detected by two dimensional difference gel electrophoresis and 274 proteins were identified byMALDITOF/TOFTM.These proteins were related to metabolic process,cellular process and response tostimulus,with mostly down-regulated in hpil leaves.Many of these proteins were related to the Calvincycle,photosynthetic electron transport chain,glycolysis/gluconeogenesis and phosphonates pathways.Some resistance-related proteins including 14-3-3 proteins,OsPR10 and antioxidases such asperoxidase,superoxide dismutase and ascorbate peroxidase were up-regulated in leaves with lesionmimic.These results provide the foundation for cloning of the target gene and shed light on themechanism involved in autaimmunity of rice.展开更多
OsPRIOa is one of the well known pathogenesis-related genes in rice,and is induced by multiple plant hormones and pathogens.However,the underlying transcriptional regulation mechanisms in response to differe nt signal...OsPRIOa is one of the well known pathogenesis-related genes in rice,and is induced by multiple plant hormones and pathogens.However,the underlying transcriptional regulation mechanisms in response to differe nt signals and their crosstalks are still largely unknown.In order to find new players participated in the activation of OsPRIOa,we systematically analyzed the basal expression patterns as well as the expression responses of a 2.5 kb OsPRIOa promoter in rice transgenic plants after phytohormone and pathogen stimulations.In agreement with the native gene expression,the OsPRIOa promoter can drive glucuronidase(GUS)gene expressing in spots of leaf cells,leaf trichomes,lemmas and paleae,germinating embryos,calli and root tips.The leaf expression of OsPR10a::GUS was dramatically in creased upon jasm onic acid(JA)and cytoki nin(CK)treatme nts,or challe nges of the pathogen Magnaporthe grisea and Xanthomonas oryzae pv.oryzae.Thus,the OsPRIOa promoter reported here can faithfully reflect its native gene expression.The effects of several JA and CK responsive OsWRKY genes on the regulation of OsPRIOa promoter were then inspected by luciferase transient expression assay,and the JA inducible OsWRKYlO transcription factor was found as a new positive regulator of OsPRIOa.However,the key transcription factors of JA and CK signaling pathways,OsMYC2 and B-type response regulators,were not responsible for the activation of OsPRIOa promote Our findings provided new in sights into the regulation of OsPRIOa expression during plan t-hormone/pathoge n interactions,and the OsPRIOa reporter system can be useful to unravel novel regulators from both pathogen and host.展开更多
基金supported by the Zhejiang Provincial Key Research and Development Plan(Grant No.2019C02006)the National Key Research and Development Program of China(Grant Nos.2016YFD0200804 and 2016YFD0100601-15)+2 种基金the Key Program of Zhejiang Provincial Foundation for Natural Science(Grant No.LZ16C130002)the Zhejiang Fundamental Public Welfare Research Program(Grant No.LGN19C140008)the State Key Laboratory for Managing Biotic and Chemical Threats to Quality and Safety of Agro-products(Grant No.2010DS700124-ZZ1907).
文摘Lesion mmic mutants(LMMs)are plants that spontaneously form lesions without pathogeninfection or external stimulus and exhibit resistance to pathogens.Here,a rice LMM was created by ethylmethane sulfonate mutagenesis,named as hpil(hydrogen peroxide induced lesion).Diaminobenzidineand trypan blue staining showed that large amounts of H_(2)O_(2) were produced and cell death was occurredat and around the parts of lesion mimic in the rice leaves.The phenotype of hpil is controlled by a singlerecessive gene,localized at a 2 Mb interval on chromosome 2.The data suggested that hpil is a novelLMM with enhanced bacterial and fungal disease resistance,and multiple pathogenesis-related proteins(PRs)were up-regulated.The proteomes of leaves at three positions(different degrees of lesion mimicseverity)were characterized in hpil compared with its wild type plant.Differentially expressed proteinswere detected by two dimensional difference gel electrophoresis and 274 proteins were identified byMALDITOF/TOFTM.These proteins were related to metabolic process,cellular process and response tostimulus,with mostly down-regulated in hpil leaves.Many of these proteins were related to the Calvincycle,photosynthetic electron transport chain,glycolysis/gluconeogenesis and phosphonates pathways.Some resistance-related proteins including 14-3-3 proteins,OsPR10 and antioxidases such asperoxidase,superoxide dismutase and ascorbate peroxidase were up-regulated in leaves with lesionmimic.These results provide the foundation for cloning of the target gene and shed light on themechanism involved in autaimmunity of rice.
基金funded by the National Key Research and Development Program of China(Grant No.2016YFD0100601-15)Zhejiang Fundamental Public Welfare Research Program(Grant No.LGN19C140008)+2 种基金Zhejiang Provincial Key Research and Development Plan(Grant No.2019C02006)Major Scientific and Technological Innovation 2025 Project of Ningbo(Grant No.2019B10004)Major Agricultural Science and Technology Project in Ningbo(Grant No.2016C11017).
文摘OsPRIOa is one of the well known pathogenesis-related genes in rice,and is induced by multiple plant hormones and pathogens.However,the underlying transcriptional regulation mechanisms in response to differe nt signals and their crosstalks are still largely unknown.In order to find new players participated in the activation of OsPRIOa,we systematically analyzed the basal expression patterns as well as the expression responses of a 2.5 kb OsPRIOa promoter in rice transgenic plants after phytohormone and pathogen stimulations.In agreement with the native gene expression,the OsPRIOa promoter can drive glucuronidase(GUS)gene expressing in spots of leaf cells,leaf trichomes,lemmas and paleae,germinating embryos,calli and root tips.The leaf expression of OsPR10a::GUS was dramatically in creased upon jasm onic acid(JA)and cytoki nin(CK)treatme nts,or challe nges of the pathogen Magnaporthe grisea and Xanthomonas oryzae pv.oryzae.Thus,the OsPRIOa promoter reported here can faithfully reflect its native gene expression.The effects of several JA and CK responsive OsWRKY genes on the regulation of OsPRIOa promoter were then inspected by luciferase transient expression assay,and the JA inducible OsWRKYlO transcription factor was found as a new positive regulator of OsPRIOa.However,the key transcription factors of JA and CK signaling pathways,OsMYC2 and B-type response regulators,were not responsible for the activation of OsPRIOa promote Our findings provided new in sights into the regulation of OsPRIOa expression during plan t-hormone/pathoge n interactions,and the OsPRIOa reporter system can be useful to unravel novel regulators from both pathogen and host.