[Objective] This study was to investigate the expression of the specific protein in Setcreasea purpurea Boom under copper stress, with the aim to clarify the copper tolerance mechanism of S. purpurea. [Method] Methods...[Objective] This study was to investigate the expression of the specific protein in Setcreasea purpurea Boom under copper stress, with the aim to clarify the copper tolerance mechanism of S. purpurea. [Method] Methods of water culture, elec- trophoresis and chromatography were used to analyze the molecular weight of the specific protein in the copper hyperaccumulator S. purpurea, as well as its expression time and the minimum copper concentration for the expression. And the specific protein was isolated and purified. [Result] Under copper stress, the minimum concentra- tion of copper to induce the expression of the specific protein from S. purpurea was 50 umol/L, and the expression time of the protein was in the 4th week with the molecular weight of 89.4 kDa. [Conclusion] The results show that the copper tolerance of S. purpurea is closely related with the expression of the specific protein.展开更多
Nitric oxide(NO),a bioactive signaling molecule,serves as an antioxidant and anti-stress agent under abiotic stress.A hydroponics experiment was conducted to investigate the effects of sodium nitroprusside(SNP),a ...Nitric oxide(NO),a bioactive signaling molecule,serves as an antioxidant and anti-stress agent under abiotic stress.A hydroponics experiment was conducted to investigate the effects of sodium nitroprusside(SNP),a NO donor,on tomato seedlings exposed to 50 μmol L-1CuCl 2.The results show that copper is primarily stored in the soluble cell sap fraction in the roots,especially after treatment with Cu+SNP treatment,which accounted for 66.2% of the total copper content.The copper concentration gradually decreased from the roots to the leaves.In the leaves,exogenous NO induces the storage of excess copper in the cell walls.Copper stress decreases the proportion of copper integrated with pectates and proteins,but exogenous NO remarkably reverses this trend.The alleviating effect of NO is blocked by hemoglobin.Thus,exogenous NO is likely involved in the regulation of the subcellular copper concentrations and its chemical forms under copper stress.Although exogenous NO inhibited the absorption and transport of excess copper to some extent,the copper accumulation in tomato seedlings significantly increased under copper stress.The use of exogenous NO to enhance copper tolerance in some plants is a promising method for copper remediation.展开更多
Sorghum is not only an important bio-energy crop but also a vital raw material for brewing.Exogenous copper affects the growth and metabolism of crops in specific ways.This study identified 8475 differentially expressed...Sorghum is not only an important bio-energy crop but also a vital raw material for brewing.Exogenous copper affects the growth and metabolism of crops in specific ways.This study identified 8475 differentially expressed genes(DEGs)by high-throughput transcriptome sequencing in the sorghum cultivar‘Jinnuoliang 2’after 24 h of treatment with 10 mM CuSO4.Using GO analysis,476 genes were functionally annotated,which were mainly related to catabolism and biosynthetic processes.Additionally,90 pathways were annotated by employing the KEGG analysis.Among them,glutathione metabolism and peroxisome were induced,while photosynthesis,photosynthesis-antenna protein,and carbon sequestration of photosynthetic organisms were inhibited.Of the DEGs,399 were identified to encode transcription factors belonging to 49 families.This study also identified a WRKY transcription factor-encoding gene SbWRKY24 from the transcriptome data.For studying its function,the relative expression levels of SbWRKY24 in roots and leaves post-treatment with different growth hormones and exposure to a variety of abiotic stresses were detected by RT-qPCR.SbWRKY24 showed treatment-and tis-sue-specific expression patterns,indicating its unique role in stress tolerance.This study lays a theoretical basis for the functional exploration of SbWRKY24,elucidating the mechanism of copper resistance,and elaborating on the stress responses in sorghum.It also guides the exploration of the molecular mechanism of copper ions inducing intracellular signal transduction pathways.展开更多
With the enhancement of copper (Cu) stress, the germination percentage of wheat seeds decreased gradually. Pretreatment with sodium hydrosulfide (NariS), hydrogen sulfide (H2S) donor alleviated the inhibitory ef...With the enhancement of copper (Cu) stress, the germination percentage of wheat seeds decreased gradually. Pretreatment with sodium hydrosulfide (NariS), hydrogen sulfide (H2S) donor alleviated the inhibitory effect of Cu stress in a dose- dependent manner; whereas little visible symptom was observed in germinating seeds and radicle tips cultured in NariS solutions. It was verified that H2S or HS- rather than other sulfur-containing components derived from NariS attribute to the potential role in promoting seed germination against Cu stress. Further studies showed that NariS could promote amylase and esterase activities, reduce Cu-induced disturbance of plasma membrane integrity in the radicle tips, and sustain lower levels of malondialdehyde and H202 in germinating seeds. Furthermore, NariS pretreatment increased activities of superoxide dismutase and catalase and decreased that of lipoxygenase, but showed no significant effect on ascorbate peroxidase. Alternatively, NariS prevented uptake of Cu and promoted the accumulation of free amino acids in seeds exposed to Cu. In addition, a rapid accumulation of endogenous H2S in seeds was observed at the early stage of germination, and higher level of H2S in NaHS-pretreated seeds. These data indicated that H2S was involved in the mechanism of germinating seeds' responses to Cu stress.展开更多
This study examined the effect of copper ions on the proliferation of hepatic stellate cells (HSCs) and the role of oxidative stress in this process in order to gain insight into the mechanism of he- patic fibrosis ...This study examined the effect of copper ions on the proliferation of hepatic stellate cells (HSCs) and the role of oxidative stress in this process in order to gain insight into the mechanism of he- patic fibrosis in Wilson's disease. LX-2 cells, a cell line of human HSCs, were cultured in vitro and treated with different agents including copper sulfate, N-acetyl cysteine (NAC) and buthionine sulfoxi- mine (BSO) for different time. The proliferation of LX-2 cells was measured by non-radioactive cell proliferation assay. Real-time PCR and Westem blotting were used to detect the mRNA and protein ex- pression of platelet-derived growth factor receptor 13 subunit (PDGFI3R), ELISA to determine the level of glutathione (GSH) and oxidized glutathione (GSSG), dichlorofluorescein assay to measure the level of reactive oxygen species (ROS), and lipid hydroperoxide assay to quantify the level of lipid peroxide (LPO). The results showed that copper sulfate over a certain concentration range could promote the pro- liferation of LX-2 cells in a time- and dose-dependent manner. The effect was most manifest when LX-2 cells were treated with copper sulfate at a concentration of 100 ~tmol/L for 24 h. Additionally, copper sulfate could dose-dependently increase the levels of ROS and LPO, and decrease the ratio of GSH/GSSG in LX-2 cells. The copper-induced increase in mRNA and protein expression of PDGF^R was significantly inhibited in LX-2 cells pre-treated with NAC, a precursor of GSH, and this phenome- non could be reversed by the intervention of BSO, an inhibitor of NAC. It was concluded that copper ions may directly stimulate the proliferation of HSCs via oxidative stress. Anti-oxidative stress therapies may help suppress the copper-induced activation and proliferation of HSCs.展开更多
An experiment was carried out to determine plant growth, mineral uptake, lipid peroxidation, antioxidative enzymes, and antioxidant of cucumber plants (Cucumis sativus L. cv. Xintaimici) under copper stress, either ...An experiment was carried out to determine plant growth, mineral uptake, lipid peroxidation, antioxidative enzymes, and antioxidant of cucumber plants (Cucumis sativus L. cv. Xintaimici) under copper stress, either ungrafted or grafted onto the rootstock (Cucurbitaficifolia). Excess Cu inhibited growth, photosynthesis, and pigment synthesis of grafted and ungrafted cucumber seedlings and significantly increased accumulation of Cu in roots besides reducing mineral uptake. Cu concentration in roots of grafted cucumber plants was significantly higher than that of ungrafted plants and obviously lower in leaves. The accumulation of reactive oxygen species (ROS) significantly increased in cucumber leaves under Cu stress and resulted in lipid peroxidation, and the levels of ROS and lipid peroxidation were greatly decreased by grafting. Activities of protective enzymes (superoxide dismutase, SOD; peroxidase, POD; catalase, CAT; ascorbate peroxidase, APX; dehydroascorbate reductase, DHAR; glutathione reductase, GR) and the contents of ascorbate and glutathione in leaves of grafted plants were significantly higher than those of ungrafted plants under Cu stress. Better performance of grafted cucumber plants were attributed to the higher ability of Cu accumulation in their roots, better nutrient status, and the effective scavenging system of ROS.展开更多
A greenhouse experiment was carried out to determine plant growth, reactive oxygen species (ROS) metabolism in roots and functions of plasma membrane (PM) and tonoplast in cucumber seedlings (Cucumis sativus L. e...A greenhouse experiment was carried out to determine plant growth, reactive oxygen species (ROS) metabolism in roots and functions of plasma membrane (PM) and tonoplast in cucumber seedlings (Cucumis sativus L. ev. Xintaimici) treated with 40 μmol L^-1 CuSO4·5H2O, which were either ungrafted or grafted onto the rootstock (Cucurbitaficifolia). Cu treatment inhibited growth, induced significant accumulation of H2O2 and led to serious lipid peroxidation in cucumber roots, and the ROS-scavenging enzymes activities in grafted seedlings roots were significantly higher than that of ungrafted plants, thus less accumulation in grafted cucumber roots induced by Cu. As a result, lipid peroxidation in roots decreased. Furthermore, the activities of H^-ATPase, H+-PPase and Ca^2+-ATPase in PM and/or tonoplast in grafted cucumber seedlings under Cu stress were obviously higher than that in ungrafted plants, resulting into higher ability in grafted plants to expulse the excess H+, promote the cytoplasm alkalinization, regulate the intracellular Ca^2+ concentration and brought the cytoplasma concentration of free Ca2+ to extremely low level under Cu stress.展开更多
The aim of the study is to determine whether grafting could improve antioxidant enzyme activities and polyamine contents in leaves of cucumber plants(Cucumis sativus L.cv.Xintaimici) under copper stress.Grafted(usi...The aim of the study is to determine whether grafting could improve antioxidant enzyme activities and polyamine contents in leaves of cucumber plants(Cucumis sativus L.cv.Xintaimici) under copper stress.Grafted(using Cucurbita ficifolia as rootstock) and ungrafted cucumber seedlings were cultured in deep flow technique(DFT) with the Cu2+ concentration of 40 μmol L-1.The results showed that on the 9th day of copper stress treatment,the contents of malondialdehyde(MDA) and hydrogen peroxide(H2O2),superoxide radical() producing rate,and electrolyte leakage percentage were significantly lower in grafted seedlings in comparison to those of the ungrafted seedlings,whereas the activities of antioxidants such as superoxide dismutase(SOD,EC 1.15.1.1),peroxidase(POD,EC 1.11.1.7),ascorbate peroxidase(APX,EC 1.11.1.11),catalase(CAT,EC 1.11.1.6),glutathione reductase(GR,EC 1.6.4.2),and monodehydroascorbate reductase(MDHAR,EC 1.6.5.4) of grafted seedlings were also significantly higher than that of ungrafted seedlings,and the contents of free proline and soluble protein of grafted seedlings were significantly higher than that of ungrafted seedlings.Cu2+ treatment increased the putrescine(Put) level and lowered the spermidine(Spd) and spermine(Spm) levels,thereby reducing the Put/(Spd + Spm) ratio in leaves of grafted and ungrafted seedlings.Grafting markedly reversed these Cu-induced effects for all three PAs and partially restored the Put/(Spd + Spm) ratio in leaves.These results suggest that grafting can enhance the tolerance of cucumber seedlings to Cu2+ stress by increasing the activities of antioxidants and the levels of endogenous Spd and Spm,decreasing the Put/(Spd + Spm) ratio and the levels of ROS,promoting free proline and soluble protein synthesis in cucumber seedling leaves.展开更多
基金Supported by the National Natural Science Foundation of China (30760021)the Natural Science Foundation of Jiangxi Province,China (0530016)the Program of Education Bureau of Jiangxi Province,China (ganjiaojizi[2007]No.151)~~
文摘[Objective] This study was to investigate the expression of the specific protein in Setcreasea purpurea Boom under copper stress, with the aim to clarify the copper tolerance mechanism of S. purpurea. [Method] Methods of water culture, elec- trophoresis and chromatography were used to analyze the molecular weight of the specific protein in the copper hyperaccumulator S. purpurea, as well as its expression time and the minimum copper concentration for the expression. And the specific protein was isolated and purified. [Result] Under copper stress, the minimum concentra- tion of copper to induce the expression of the specific protein from S. purpurea was 50 umol/L, and the expression time of the protein was in the 4th week with the molecular weight of 89.4 kDa. [Conclusion] The results show that the copper tolerance of S. purpurea is closely related with the expression of the specific protein.
基金supported partially by the National Natural Science Foundation of China (31201619)Profession Expert Group of Facility Cultivation and Engineering (CARS25-D-03)the Sci-Tech Development Project of Tai’an City, China (32606)
文摘Nitric oxide(NO),a bioactive signaling molecule,serves as an antioxidant and anti-stress agent under abiotic stress.A hydroponics experiment was conducted to investigate the effects of sodium nitroprusside(SNP),a NO donor,on tomato seedlings exposed to 50 μmol L-1CuCl 2.The results show that copper is primarily stored in the soluble cell sap fraction in the roots,especially after treatment with Cu+SNP treatment,which accounted for 66.2% of the total copper content.The copper concentration gradually decreased from the roots to the leaves.In the leaves,exogenous NO induces the storage of excess copper in the cell walls.Copper stress decreases the proportion of copper integrated with pectates and proteins,but exogenous NO remarkably reverses this trend.The alleviating effect of NO is blocked by hemoglobin.Thus,exogenous NO is likely involved in the regulation of the subcellular copper concentrations and its chemical forms under copper stress.Although exogenous NO inhibited the absorption and transport of excess copper to some extent,the copper accumulation in tomato seedlings significantly increased under copper stress.The use of exogenous NO to enhance copper tolerance in some plants is a promising method for copper remediation.
基金funded by the Key Planned Projects of the Sichuan Provincial Department of Science&Technology(2020YFN0023)the Cooperation Project of Wuliangye Group Co.,Ltd.,and Sichuan University of Science&Engineering,China(CXY2021ZR010).
文摘Sorghum is not only an important bio-energy crop but also a vital raw material for brewing.Exogenous copper affects the growth and metabolism of crops in specific ways.This study identified 8475 differentially expressed genes(DEGs)by high-throughput transcriptome sequencing in the sorghum cultivar‘Jinnuoliang 2’after 24 h of treatment with 10 mM CuSO4.Using GO analysis,476 genes were functionally annotated,which were mainly related to catabolism and biosynthetic processes.Additionally,90 pathways were annotated by employing the KEGG analysis.Among them,glutathione metabolism and peroxisome were induced,while photosynthesis,photosynthesis-antenna protein,and carbon sequestration of photosynthetic organisms were inhibited.Of the DEGs,399 were identified to encode transcription factors belonging to 49 families.This study also identified a WRKY transcription factor-encoding gene SbWRKY24 from the transcriptome data.For studying its function,the relative expression levels of SbWRKY24 in roots and leaves post-treatment with different growth hormones and exposure to a variety of abiotic stresses were detected by RT-qPCR.SbWRKY24 showed treatment-and tis-sue-specific expression patterns,indicating its unique role in stress tolerance.This study lays a theoretical basis for the functional exploration of SbWRKY24,elucidating the mechanism of copper resistance,and elaborating on the stress responses in sorghum.It also guides the exploration of the molecular mechanism of copper ions inducing intracellular signal transduction pathways.
基金Supported by the Natural Science Foundation of Anhui Province (070411009)the Innovation Fund from Hefei University of Technology to undergraduate students (XS0637).
文摘With the enhancement of copper (Cu) stress, the germination percentage of wheat seeds decreased gradually. Pretreatment with sodium hydrosulfide (NariS), hydrogen sulfide (H2S) donor alleviated the inhibitory effect of Cu stress in a dose- dependent manner; whereas little visible symptom was observed in germinating seeds and radicle tips cultured in NariS solutions. It was verified that H2S or HS- rather than other sulfur-containing components derived from NariS attribute to the potential role in promoting seed germination against Cu stress. Further studies showed that NariS could promote amylase and esterase activities, reduce Cu-induced disturbance of plasma membrane integrity in the radicle tips, and sustain lower levels of malondialdehyde and H202 in germinating seeds. Furthermore, NariS pretreatment increased activities of superoxide dismutase and catalase and decreased that of lipoxygenase, but showed no significant effect on ascorbate peroxidase. Alternatively, NariS prevented uptake of Cu and promoted the accumulation of free amino acids in seeds exposed to Cu. In addition, a rapid accumulation of endogenous H2S in seeds was observed at the early stage of germination, and higher level of H2S in NaHS-pretreated seeds. These data indicated that H2S was involved in the mechanism of germinating seeds' responses to Cu stress.
文摘This study examined the effect of copper ions on the proliferation of hepatic stellate cells (HSCs) and the role of oxidative stress in this process in order to gain insight into the mechanism of he- patic fibrosis in Wilson's disease. LX-2 cells, a cell line of human HSCs, were cultured in vitro and treated with different agents including copper sulfate, N-acetyl cysteine (NAC) and buthionine sulfoxi- mine (BSO) for different time. The proliferation of LX-2 cells was measured by non-radioactive cell proliferation assay. Real-time PCR and Westem blotting were used to detect the mRNA and protein ex- pression of platelet-derived growth factor receptor 13 subunit (PDGFI3R), ELISA to determine the level of glutathione (GSH) and oxidized glutathione (GSSG), dichlorofluorescein assay to measure the level of reactive oxygen species (ROS), and lipid hydroperoxide assay to quantify the level of lipid peroxide (LPO). The results showed that copper sulfate over a certain concentration range could promote the pro- liferation of LX-2 cells in a time- and dose-dependent manner. The effect was most manifest when LX-2 cells were treated with copper sulfate at a concentration of 100 ~tmol/L for 24 h. Additionally, copper sulfate could dose-dependently increase the levels of ROS and LPO, and decrease the ratio of GSH/GSSG in LX-2 cells. The copper-induced increase in mRNA and protein expression of PDGF^R was significantly inhibited in LX-2 cells pre-treated with NAC, a precursor of GSH, and this phenome- non could be reversed by the intervention of BSO, an inhibitor of NAC. It was concluded that copper ions may directly stimulate the proliferation of HSCs via oxidative stress. Anti-oxidative stress therapies may help suppress the copper-induced activation and proliferation of HSCs.
基金supported by the Young Scientist Innovation Science of Shandong Agricultural University,China (23653)
文摘An experiment was carried out to determine plant growth, mineral uptake, lipid peroxidation, antioxidative enzymes, and antioxidant of cucumber plants (Cucumis sativus L. cv. Xintaimici) under copper stress, either ungrafted or grafted onto the rootstock (Cucurbitaficifolia). Excess Cu inhibited growth, photosynthesis, and pigment synthesis of grafted and ungrafted cucumber seedlings and significantly increased accumulation of Cu in roots besides reducing mineral uptake. Cu concentration in roots of grafted cucumber plants was significantly higher than that of ungrafted plants and obviously lower in leaves. The accumulation of reactive oxygen species (ROS) significantly increased in cucumber leaves under Cu stress and resulted in lipid peroxidation, and the levels of ROS and lipid peroxidation were greatly decreased by grafting. Activities of protective enzymes (superoxide dismutase, SOD; peroxidase, POD; catalase, CAT; ascorbate peroxidase, APX; dehydroascorbate reductase, DHAR; glutathione reductase, GR) and the contents of ascorbate and glutathione in leaves of grafted plants were significantly higher than those of ungrafted plants under Cu stress. Better performance of grafted cucumber plants were attributed to the higher ability of Cu accumulation in their roots, better nutrient status, and the effective scavenging system of ROS.
基金financially supported by Modern Agricultural Industurl Techenology System Fund of Shandong Province,China (2010-03)
文摘A greenhouse experiment was carried out to determine plant growth, reactive oxygen species (ROS) metabolism in roots and functions of plasma membrane (PM) and tonoplast in cucumber seedlings (Cucumis sativus L. ev. Xintaimici) treated with 40 μmol L^-1 CuSO4·5H2O, which were either ungrafted or grafted onto the rootstock (Cucurbitaficifolia). Cu treatment inhibited growth, induced significant accumulation of H2O2 and led to serious lipid peroxidation in cucumber roots, and the ROS-scavenging enzymes activities in grafted seedlings roots were significantly higher than that of ungrafted plants, thus less accumulation in grafted cucumber roots induced by Cu. As a result, lipid peroxidation in roots decreased. Furthermore, the activities of H^-ATPase, H+-PPase and Ca^2+-ATPase in PM and/or tonoplast in grafted cucumber seedlings under Cu stress were obviously higher than that in ungrafted plants, resulting into higher ability in grafted plants to expulse the excess H+, promote the cytoplasm alkalinization, regulate the intracellular Ca^2+ concentration and brought the cytoplasma concentration of free Ca2+ to extremely low level under Cu stress.
基金supported by the Young Scientist Innovation Science of Shandong Agricultural University,China (23653)
文摘The aim of the study is to determine whether grafting could improve antioxidant enzyme activities and polyamine contents in leaves of cucumber plants(Cucumis sativus L.cv.Xintaimici) under copper stress.Grafted(using Cucurbita ficifolia as rootstock) and ungrafted cucumber seedlings were cultured in deep flow technique(DFT) with the Cu2+ concentration of 40 μmol L-1.The results showed that on the 9th day of copper stress treatment,the contents of malondialdehyde(MDA) and hydrogen peroxide(H2O2),superoxide radical() producing rate,and electrolyte leakage percentage were significantly lower in grafted seedlings in comparison to those of the ungrafted seedlings,whereas the activities of antioxidants such as superoxide dismutase(SOD,EC 1.15.1.1),peroxidase(POD,EC 1.11.1.7),ascorbate peroxidase(APX,EC 1.11.1.11),catalase(CAT,EC 1.11.1.6),glutathione reductase(GR,EC 1.6.4.2),and monodehydroascorbate reductase(MDHAR,EC 1.6.5.4) of grafted seedlings were also significantly higher than that of ungrafted seedlings,and the contents of free proline and soluble protein of grafted seedlings were significantly higher than that of ungrafted seedlings.Cu2+ treatment increased the putrescine(Put) level and lowered the spermidine(Spd) and spermine(Spm) levels,thereby reducing the Put/(Spd + Spm) ratio in leaves of grafted and ungrafted seedlings.Grafting markedly reversed these Cu-induced effects for all three PAs and partially restored the Put/(Spd + Spm) ratio in leaves.These results suggest that grafting can enhance the tolerance of cucumber seedlings to Cu2+ stress by increasing the activities of antioxidants and the levels of endogenous Spd and Spm,decreasing the Put/(Spd + Spm) ratio and the levels of ROS,promoting free proline and soluble protein synthesis in cucumber seedling leaves.