H_2O_2和NO互作调控SO_2诱导的胡杨细胞死亡
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摘要
二氧化硫(SO_2)是一种常见的大气污染物,目前关于SO_2对木本植物的毒害作用及相关机制并不清楚.本文以木本植物胡杨的愈伤细胞为材料,研究SO_2衍生物对胡杨细胞的致死效应,以及过氧化氢(H_2O_2)与一氧化氮(NO)在SO_2诱导胡杨细胞死亡中的信号调节作用.研究发现:SO_2衍生物处理(1~5 mmol·L-1)可诱发胡杨细胞死亡,且SO_2衍生物浓度越大、处理时间越长,细胞死亡率越高.2 mmol·L-1SO_2衍生物处理胡杨细胞后,胞内H_2O_2和NO水平显著升高,且H_2O_2水平的升高先于NO.一定浓度的外源H_2O_2或NO供体SNP能够提高SO_2胁迫下胡杨细胞的死亡率;而使用H_2O_2清除剂CAT和ASA、NO清除剂c PTIO、NO合成抑制剂钨酸钠后,SO_2诱导的细胞死亡率明显降低.进一步实验发现,外源H_2O_2可以提高SO_2胁迫下胡杨细胞的硝酸还原酶(NR)活性,促进胞内NO产生;而利用CAT和ASA清除H_2O_2后,细胞NR活性和NO产生均受到明显抑制.此外,SO_2胁迫下,外源SNP能够抑制抗氧化酶(CAT和APX)活性,增加胡杨细胞内的H_2O_2水平,而一定浓度的c PTIO和钨酸钠均可提高CAT和APX活性,降低胞内H_2O_2水平.结果表明:SO_2胁迫下,胡杨细胞快速产生的H_2O_2能够激活硝酸还原酶活性,促进NO生成,同时NO能够通过抑制抗氧化酶活性而提高H_2O_2水平.H_2O_2与NO互作调控SO_2诱导的胡杨细胞死亡.
Sulfur dioxide(SO_2) is a common air pollutant.In woody plants,the toxic effects of SO_2 on cells and the relevant mechanisms remain largely unknown.In this study,callus cells of a woody species Populus euphratica were applied to investigate the lethal effects of SO_2 and the roles of H_2O_2 and NO in the regulation of SO_2-induced cell death.The results showed that 1 ~ 5 mmol·L-1SO_2 derivatives caused a dose-and time-dependent increase of cell death rate.Upon 2 mmol·L-1SO_2 treatment,the intercellular H_2O_2 and NO levels increased,and the rise of H_2O_2 level was prior to NO.Exogenous H_2O_2 or NO donor elevated SO_2-induced cell death rate,while application of H_2O_2scavenger(catalase(CAT) or ascorbic acid(ASA)),NO scavenger(c PTIO),or NO synthase inhibitor(sodium tungstate) markedly reduced the cell death rate induced by SO_2.Further studies showed that exogenous H_2O_2 enhanced nitrate reductase(NR) activity and promoted NO production in P.euphratica cells exposure to SO_2 stress.When CAT or ASA was applied,the NR activity and SO_2-induced NO generation was significantly repressed.Meanwhile,exogenous SNP reduced the antioxidant enzymes activities(CAT and APX),and thus enhanced intercellular H_2O_2level;whereas c PTIO and sodium tungstate led to increased CAT and APX activities and decreased H_2O_2 accumulation within cells.These results suggested that in SO_2-stressed P.euphratica cells,H_2O_2 could activate nitrate reductase activity and thus promote NO generation.NO could improve H_2O_2 accumulation by inhibiting antioxidant enzymes activities.Crosstalk between H_2O_2 and NO regulated the SO_2-induced cell death of P.euphratica.
Sulfur dioxide(SO_2) is a common air pollutant.In woody plants,the toxic effects of SO_2 on cells and the relevant mechanisms remain largely unknown.In this study,callus cells of a woody species Populus euphratica were applied to investigate the lethal effects of SO_2 and the roles of H_2O_2 and NO in the regulation of SO_2-induced cell death.The results showed that 1 ~ 5 mmol·L-1SO_2 derivatives caused a dose-and time-dependent increase of cell death rate.Upon 2 mmol·L-1SO_2 treatment,the intercellular H_2O_2 and NO levels increased,and the rise of H_2O_2 level was prior to NO.Exogenous H_2O_2 or NO donor elevated SO_2-induced cell death rate,while application of H_2O_2scavenger(catalase(CAT) or ascorbic acid(ASA)),NO scavenger(c PTIO),or NO synthase inhibitor(sodium tungstate) markedly reduced the cell death rate induced by SO_2.Further studies showed that exogenous H_2O_2 enhanced nitrate reductase(NR) activity and promoted NO production in P.euphratica cells exposure to SO_2 stress.When CAT or ASA was applied,the NR activity and SO_2-induced NO generation was significantly repressed.Meanwhile,exogenous SNP reduced the antioxidant enzymes activities(CAT and APX),and thus enhanced intercellular H_2O_2level;whereas c PTIO and sodium tungstate led to increased CAT and APX activities and decreased H_2O_2 accumulation within cells.These results suggested that in SO_2-stressed P.euphratica cells,H_2O_2 could activate nitrate reductase activity and thus promote NO generation.NO could improve H_2O_2 accumulation by inhibiting antioxidant enzymes activities.Crosstalk between H_2O_2 and NO regulated the SO_2-induced cell death of P.euphratica.
引文
Brychkova G,Xia Z,Yang G,et al.2007.Sulfite oxidase protects plant against sulfur dioxide toxicity[J].The Plant Journal,50(4):696-709
Bright J,Desikan R,Hancock J T,et al.2006.ABA-induced NOgeneration and stomatal closure in Arabidopsis are dependent on H2O2synthesis[J].The Plant Journal,45(1):113-122
Choi D,Toda H,Kim Y,et al.2014.Effect of sulfur dioxide(SO2)on growth and physiological activity in Alnus sieboldiana at Miyakejim Island in Japan[J].Ecological Research,29(1):103-110
De Michele R,Vurro E,Rigo C,et al.2009.Nitric oxide is involved in cadmium-induced programmed cell death in Arabidopsis suspension cultures[J].Plant Physiology,150(1):217-228
Disikan R,Griffiths R,Hancock J,et al.2002.A new role for an old enzyme:Nitrate reductase-mediated nitric oxide generation is required for abscisic acid-induced stomatal closure in Arabidopsis thaliana[J].Proceedings of the National Academy of Sciences,99(25):16314-16318
付宝春,魏爱丽,翟晓燕,等.2015.NO、ROS对诱导的万寿菊保卫细胞凋亡的调控[J].环境科学学报,35(7):2289-2296
Lang C,Popko J,Wirtz M,et al.2007.Sulfite oxidase as key enzyme fo protecting plants against sulfur dioxide[J].Plant,Cell and Environment,30(4):447-455
Lewandowska M,Sirko A.2008.Recent advances in understanding plan response to sulfur-deficiency stress[J].Acta Biochimica Polonica,55(3):457-471
Li L,Yi H.2012.Differential expression of Arabidopsis defense-related genes in response to sulfur dioxide[J].Chemosphere,87(7):718-724.
李蕊,仪慧兰,仪民.2015.一氧化氮合酶途径参与SO2胁迫下蚕豆气孔运动调节[J].环境科学学报,35(10):3406-3410
刘洁,王省芬,张桂寅,等.2010.棉花叶片硝酸还原酶活性的测定方法[J].河北农业大学学报,33(4):1-4
Lu Z,Streets D G,Zhang Q,et al.2010.Sulfur dioxide emissions in Chin and sulfur trends in East Asia since 2000[J].Atmospheric Chemistr and Physics,10(13):6311-6331
Ma W,Xu W,Xu H,et al.2010.Nitric oxide modulates cadmium influ during cadmium-induced programmed cell death in tobacco BY-cells[J].Planta,232(2):325-335
Qiao W,Li C,Fan L.2014.Cross-talk between nitric oxide and hydrogen peroxide in plant responses to abiotic stresses[J].Environmental and Experimental Botany 100:84-93
Randewig D,Hamisch D,Herschbach C,et al.2012.Sulfite oxidas controls sulfur metabolism under SO2exposure in Arabidopsi thaliana[J].Plant,Cell and Environment,35(1):100-115
Shapiro R.1977.Genetic effects of bisulfite[J].Mutation Research,39149-176
Sun J,Li L,Liu M,et al.2010.Hydrogen peroxide and nitric oxide mediat K+/Na+homeostasis and antioxidant defense in NaC l-stressed callu cells of two contrasting poplars[J].Plant Cell,Tissue and Organ Culture,103(2):205-215
Sun J,Zhan C,Deng S,et al.2012.An ATP signalling pathway in plan cells:extracellular ATP triggers programmed cell death in Populu euphratica[J].Plant,Cell and Environment,35(5):893-916
Sun J,Yu Y,Li Z,et al.2014.1-Butanol triggers programmed cell death in Populus euphratica cell cultures[J].Plant Growth Regulation,74(1):33-45
Wang Y,Loake GJ,Chu C.2013.Cross-talk of nitric oxide and reactiv oxygen species in plant programed cell death[J].Frontiers in Plan Science,4:314
Wang L,Guo Y,Jia L,et al.2014.Hydrogen peroxide acts upstream o nitric oxide in the heat shock pathway in Arabidopsis seedlings[J]Plant Physiology,164(4):2184-2196
Wei A,Xin X,Wang Y,et al.2013.Signal regulation involved in sulfu dioxide-induced guard cell apoptosis in Hemerocallis fulva[J]Ecotoxicology and Environmental Safety,98(1):41-45
Wei A,Fu B,Wang Y,et al.2015.Involvement of NO and ROS in sulfu dioxide induced guard cells apoptosis in Tagetes erecta[J]Ecotoxicology and Environmental Safety,114:198-203
Yi H,Yin J,Liu X,et al.2012.Sulfur dioxide induced programmed cel death in Vicia guard cells[J].Ecotoxicology and Environmenta Safety,78(1):281-286
尹晶晶,刘鑫,仪慧兰,等.2010.二氧化硫诱导蚕豆气孔保卫细胞死亡效应研究[J].环境科学学报,30(12):2512-2517
赵均,薛美昭,白鹤丽,等.2014.硝酸还原酶介导的NO合成参于SO2引发的拟南芥气孔运动的调节[J].环境科学学报,34(3)796-800
Bright J,Desikan R,Hancock J T,et al.2006.ABA-induced NOgeneration and stomatal closure in Arabidopsis are dependent on H2O2synthesis[J].The Plant Journal,45(1):113-122
Choi D,Toda H,Kim Y,et al.2014.Effect of sulfur dioxide(SO2)on growth and physiological activity in Alnus sieboldiana at Miyakejim Island in Japan[J].Ecological Research,29(1):103-110
De Michele R,Vurro E,Rigo C,et al.2009.Nitric oxide is involved in cadmium-induced programmed cell death in Arabidopsis suspension cultures[J].Plant Physiology,150(1):217-228
Disikan R,Griffiths R,Hancock J,et al.2002.A new role for an old enzyme:Nitrate reductase-mediated nitric oxide generation is required for abscisic acid-induced stomatal closure in Arabidopsis thaliana[J].Proceedings of the National Academy of Sciences,99(25):16314-16318
付宝春,魏爱丽,翟晓燕,等.2015.NO、ROS对诱导的万寿菊保卫细胞凋亡的调控[J].环境科学学报,35(7):2289-2296
Lang C,Popko J,Wirtz M,et al.2007.Sulfite oxidase as key enzyme fo protecting plants against sulfur dioxide[J].Plant,Cell and Environment,30(4):447-455
Lewandowska M,Sirko A.2008.Recent advances in understanding plan response to sulfur-deficiency stress[J].Acta Biochimica Polonica,55(3):457-471
Li L,Yi H.2012.Differential expression of Arabidopsis defense-related genes in response to sulfur dioxide[J].Chemosphere,87(7):718-724.
李蕊,仪慧兰,仪民.2015.一氧化氮合酶途径参与SO2胁迫下蚕豆气孔运动调节[J].环境科学学报,35(10):3406-3410
刘洁,王省芬,张桂寅,等.2010.棉花叶片硝酸还原酶活性的测定方法[J].河北农业大学学报,33(4):1-4
Lu Z,Streets D G,Zhang Q,et al.2010.Sulfur dioxide emissions in Chin and sulfur trends in East Asia since 2000[J].Atmospheric Chemistr and Physics,10(13):6311-6331
Ma W,Xu W,Xu H,et al.2010.Nitric oxide modulates cadmium influ during cadmium-induced programmed cell death in tobacco BY-cells[J].Planta,232(2):325-335
Qiao W,Li C,Fan L.2014.Cross-talk between nitric oxide and hydrogen peroxide in plant responses to abiotic stresses[J].Environmental and Experimental Botany 100:84-93
Randewig D,Hamisch D,Herschbach C,et al.2012.Sulfite oxidas controls sulfur metabolism under SO2exposure in Arabidopsi thaliana[J].Plant,Cell and Environment,35(1):100-115
Shapiro R.1977.Genetic effects of bisulfite[J].Mutation Research,39149-176
Sun J,Li L,Liu M,et al.2010.Hydrogen peroxide and nitric oxide mediat K+/Na+homeostasis and antioxidant defense in NaC l-stressed callu cells of two contrasting poplars[J].Plant Cell,Tissue and Organ Culture,103(2):205-215
Sun J,Zhan C,Deng S,et al.2012.An ATP signalling pathway in plan cells:extracellular ATP triggers programmed cell death in Populu euphratica[J].Plant,Cell and Environment,35(5):893-916
Sun J,Yu Y,Li Z,et al.2014.1-Butanol triggers programmed cell death in Populus euphratica cell cultures[J].Plant Growth Regulation,74(1):33-45
Wang Y,Loake GJ,Chu C.2013.Cross-talk of nitric oxide and reactiv oxygen species in plant programed cell death[J].Frontiers in Plan Science,4:314
Wang L,Guo Y,Jia L,et al.2014.Hydrogen peroxide acts upstream o nitric oxide in the heat shock pathway in Arabidopsis seedlings[J]Plant Physiology,164(4):2184-2196
Wei A,Xin X,Wang Y,et al.2013.Signal regulation involved in sulfu dioxide-induced guard cell apoptosis in Hemerocallis fulva[J]Ecotoxicology and Environmental Safety,98(1):41-45
Wei A,Fu B,Wang Y,et al.2015.Involvement of NO and ROS in sulfu dioxide induced guard cells apoptosis in Tagetes erecta[J]Ecotoxicology and Environmental Safety,114:198-203
Yi H,Yin J,Liu X,et al.2012.Sulfur dioxide induced programmed cel death in Vicia guard cells[J].Ecotoxicology and Environmenta Safety,78(1):281-286
尹晶晶,刘鑫,仪慧兰,等.2010.二氧化硫诱导蚕豆气孔保卫细胞死亡效应研究[J].环境科学学报,30(12):2512-2517
赵均,薛美昭,白鹤丽,等.2014.硝酸还原酶介导的NO合成参于SO2引发的拟南芥气孔运动的调节[J].环境科学学报,34(3)796-800