Hydrogen peroxide （H2O2） is an important signaling molecule in ethylene-induced stomatal closure in Arabidopsis thaliana. Early studies on the sources of H2O2 mainly focused on NADPH oxidases and cell-wall peroxidas...Hydrogen peroxide （H2O2） is an important signaling molecule in ethylene-induced stomatal closure in Arabidopsis thaliana. Early studies on the sources of H2O2 mainly focused on NADPH oxidases and cell-wall peroxidases. Here, we report the involvement of polyamine oxidases （PAOs） in ethylene-induced H2O2 production in guard cells. In Arabidopsis epidermal peels, application of PAO inhibitors caused the failure of ethylene to induce H2O2 production and stomatal closure. Results of quantitative RT-PCR analysis and pharmacological experiments showed that AtPAO2 and AtPAO4 transcripts and activities of PAOs were both induced by ethylene. In transgenic Arabidopsis plants over-expressing AtPAO2 and AtPAO4, stomatal movement was more sensitive to ethylene treatment and H2O2 production was also significantly induced. The increased H2O2 production in the transgenic lines compared to the wild-type plants suggests that AtPAO2 and AtPAO4 probably are involved in ethylene-induced H2O2 production. Several factors which induce stomatal closure such as dehydration and high salinity all enhanced the expression of AtPAO2 and AtPAO4 to different degrees. Moreover, GFP- AtPAOs fusion protein localized in the nucleus, cytoplasm, and cell wall of the guard cells. Therefore, our results strongly indicated that PAO is a source of H2O2 generation in Arabidopsis guard cells and plays crucial roles in stomatal movement.展开更多
Oxalic acid（OA） is considered as an important pathogenetic factor of some destructive diseases caused by some fungal pathogens such as Sclerotinia sclerotiorum. Oxalate degradation is important for plant health, and...Oxalic acid（OA） is considered as an important pathogenetic factor of some destructive diseases caused by some fungal pathogens such as Sclerotinia sclerotiorum. Oxalate degradation is important for plant health, and plants that contain oxalate oxidase（OXO） enzymes could breakdown oxalate into CO_2 and H_2O_2, which subsequently evokes defense responses. However, some species, such as Arabidopsis thaliana, have no oxalate oxidase activity identified to date. The present study aims to develop transgenic Arabidopsis expressing a wheat oxalate oxidase, to test for the response to OA exposure and fungal infection by S. sclerotiorum. The results showed that the transgenic Arabidopsis lines that expressed the wheat OXO exhibited enhanced resistance to OA exposure and S. sclerotiorum infection in the tolerance assays. In the same manner, it could convert OA to CO_2 and H_2O_2 to a higher extent than the wild-type. Intensive osmotic adjustments were also detected in the transgenic Arabidopsis lines. The higher level of produced H_2O_2 subsequently induced an elevated activity of antioxidant enzymes including superoxide dismutase（SOD） and peroxidase（POD） in the transgenic Arabidopsis plants. The present study indicated that the expression of a gene encoding wheat OXO could induce intensive osmotic adjustments and hydrogen peroxide related defense response, and subsequently increased tolerance to S. sclerotiorum in transgenic A. thaliana.展开更多
In non-cyanogenic species, the main source of cyanide derives from ethylene and camalexin biosyntheses. In mitochondria, cyanide is a potent inhibitor of the cytochrome c oxidase and is metabolized by the β-cyanoalan...In non-cyanogenic species, the main source of cyanide derives from ethylene and camalexin biosyntheses. In mitochondria, cyanide is a potent inhibitor of the cytochrome c oxidase and is metabolized by the β-cyanoalanine synthase CYS-C1, catalyzing the conversion of cysteine and cyanide to hydrogen sulfide and β-cyanoalanine. The hydrogen sulfide released also inhibits the cytochrome c oxidase and needs to be detoxified by the O-acetylserine（thiol）lyase mitochondrial isoform, OAS-C, which catalyzes the incorporation of sulfide to O-acetylserine to produce cysteine, thus generating a cyclic pathway in the mitochondria. The loss of functional OAS-C isoforms causes phenotypic characteristics very similar to the loss of the CYS-C1 enzyme, showing defects in root hair formation. Genetic complementation with the OAS-Cgene rescues the impairment of root hair elongation, restoring the wild-type phenotype. The mitochondria compromise their capacity to properly detoxify cyanide and the resulting sulfide because the latter cannot re-assimilate into cysteine in the oas-c null mutant. Consequently, we observe an accumulation of sulfide and cyanide and of the alternative oxidase, which is unable to prevent the production of reactive oxygen species probably due to the accumulation of both toxic molecules. Our results allow us to suggest that the significance of OAS-C is related to its role in the proper sulfide and cyanide detoxification in mitochondria.展开更多
The three dimensional structure model for protoporphyrinogen oxidase (Arabidopsis thaliana) has been established theoretically,using comparative protein modeling by satisfaction of spatial restrains and fold recogniti...The three dimensional structure model for protoporphyrinogen oxidase (Arabidopsis thaliana) has been established theoretically,using comparative protein modeling by satisfaction of spatial restrains and fold recognition methods,and the loops structures restrans were obtained by scanning the PDB data bank.The last optimized model shows adequate fitting to protoporphyrinogen Ⅸ.展开更多
基金supported by the National Natural Science Foundation of China(30970228 and 31170237)the National Science Foundation of Shandong Province,China(ZR2010CM024)the Foundation of State Key Laboratory of Plant Physiology and Biochemistry,China(SKLPPBKF11001)
文摘Hydrogen peroxide （H2O2） is an important signaling molecule in ethylene-induced stomatal closure in Arabidopsis thaliana. Early studies on the sources of H2O2 mainly focused on NADPH oxidases and cell-wall peroxidases. Here, we report the involvement of polyamine oxidases （PAOs） in ethylene-induced H2O2 production in guard cells. In Arabidopsis epidermal peels, application of PAO inhibitors caused the failure of ethylene to induce H2O2 production and stomatal closure. Results of quantitative RT-PCR analysis and pharmacological experiments showed that AtPAO2 and AtPAO4 transcripts and activities of PAOs were both induced by ethylene. In transgenic Arabidopsis plants over-expressing AtPAO2 and AtPAO4, stomatal movement was more sensitive to ethylene treatment and H2O2 production was also significantly induced. The increased H2O2 production in the transgenic lines compared to the wild-type plants suggests that AtPAO2 and AtPAO4 probably are involved in ethylene-induced H2O2 production. Several factors which induce stomatal closure such as dehydration and high salinity all enhanced the expression of AtPAO2 and AtPAO4 to different degrees. Moreover, GFP- AtPAOs fusion protein localized in the nucleus, cytoplasm, and cell wall of the guard cells. Therefore, our results strongly indicated that PAO is a source of H2O2 generation in Arabidopsis guard cells and plays crucial roles in stomatal movement.
基金financially supported by the National Key Technology R&D Program of China(2010BAD01B02)the National Natural Science Foundation of China(U1204308)the Education Department of Henan Province,China(13A180437)
文摘Oxalic acid（OA） is considered as an important pathogenetic factor of some destructive diseases caused by some fungal pathogens such as Sclerotinia sclerotiorum. Oxalate degradation is important for plant health, and plants that contain oxalate oxidase（OXO） enzymes could breakdown oxalate into CO_2 and H_2O_2, which subsequently evokes defense responses. However, some species, such as Arabidopsis thaliana, have no oxalate oxidase activity identified to date. The present study aims to develop transgenic Arabidopsis expressing a wheat oxalate oxidase, to test for the response to OA exposure and fungal infection by S. sclerotiorum. The results showed that the transgenic Arabidopsis lines that expressed the wheat OXO exhibited enhanced resistance to OA exposure and S. sclerotiorum infection in the tolerance assays. In the same manner, it could convert OA to CO_2 and H_2O_2 to a higher extent than the wild-type. Intensive osmotic adjustments were also detected in the transgenic Arabidopsis lines. The higher level of produced H_2O_2 subsequently induced an elevated activity of antioxidant enzymes including superoxide dismutase（SOD） and peroxidase（POD） in the transgenic Arabidopsis plants. The present study indicated that the expression of a gene encoding wheat OXO could induce intensive osmotic adjustments and hydrogen peroxide related defense response, and subsequently increased tolerance to S. sclerotiorum in transgenic A. thaliana.
文摘In non-cyanogenic species, the main source of cyanide derives from ethylene and camalexin biosyntheses. In mitochondria, cyanide is a potent inhibitor of the cytochrome c oxidase and is metabolized by the β-cyanoalanine synthase CYS-C1, catalyzing the conversion of cysteine and cyanide to hydrogen sulfide and β-cyanoalanine. The hydrogen sulfide released also inhibits the cytochrome c oxidase and needs to be detoxified by the O-acetylserine（thiol）lyase mitochondrial isoform, OAS-C, which catalyzes the incorporation of sulfide to O-acetylserine to produce cysteine, thus generating a cyclic pathway in the mitochondria. The loss of functional OAS-C isoforms causes phenotypic characteristics very similar to the loss of the CYS-C1 enzyme, showing defects in root hair formation. Genetic complementation with the OAS-Cgene rescues the impairment of root hair elongation, restoring the wild-type phenotype. The mitochondria compromise their capacity to properly detoxify cyanide and the resulting sulfide because the latter cannot re-assimilate into cysteine in the oas-c null mutant. Consequently, we observe an accumulation of sulfide and cyanide and of the alternative oxidase, which is unable to prevent the production of reactive oxygen species probably due to the accumulation of both toxic molecules. Our results allow us to suggest that the significance of OAS-C is related to its role in the proper sulfide and cyanide detoxification in mitochondria.
文摘The three dimensional structure model for protoporphyrinogen oxidase (Arabidopsis thaliana) has been established theoretically,using comparative protein modeling by satisfaction of spatial restrains and fold recognition methods,and the loops structures restrans were obtained by scanning the PDB data bank.The last optimized model shows adequate fitting to protoporphyrinogen Ⅸ.
