硫代腺苷甲硫氨酸促进番茄百菌清降解的生理机制
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摘要
【目的】探讨硫代腺苷甲硫氨酸(SAM)对番茄农药代谢调控的生理机制,为蔬菜安全生产奠定理论基础。【方法】本研究选取鲜食樱桃番茄‘千禧’为供试植物,以生产上广泛应用的广谱性杀菌剂‘百菌清’为供试农药,研究外源SAM对番茄果实喷施百菌清后的农药残留量、活性氧含量和谷胱甘肽代谢系统的影响。【结果】外源喷施0.5—2μmol·L~(-1)的SAM均能显著降低番茄果实的百菌清残留量;但当外源SAM处理浓度达到0.5μmol·L~(-1)后,番茄果实中的百菌清残留量并未随着处理浓度的增加而下降。与对照相比,喷施百菌清后番茄果实的谷胱甘肽S-转移酶(GST)、谷胱甘肽还原酶(GR)和脱氢抗坏血酸还原酶(DHAR)活性显著上升;还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)和总谷胱甘肽含量(GSH+GSSG)变化也呈现上升趋势,谷胱甘肽的还原氧化比(GSH/GSSG)呈先上升后下降趋势;超氧阴离子(O_2~-)和过氧化氢(H_2O_2)含量显著上升,而丙二醛(MDA)含量与对照相比差异不显著。外源喷施0.5μmol·L~(-1)的SAM可显著提高百菌清处理下GST、GR和DHAR的活性,增加GSH和GSH+GSSG含量,降低处理10 d后的植株GSSG含量,提高GSH/GSSG;提高处理2—5 d的植株?和H_2O_2含量,但对MDA含量无显著影响。【结论】外源添加0.5μmol·L~(-1)的SAM可激活谷胱甘肽循环系统关键酶GR和DHAR活性,促进GSH再生,依赖GST并协同?和H_2O_2的氧化作用,促进番茄果实中的百菌清降解。
【Objective】 In order to provide basic mechanism for vegetable safe production, this study aimed at revealing the physiological mechanism of S-adenosylmethionine(SAM)-induced pesticide metabolism. 【Method】 The fresh-eating tomato‘QianXi' and broad-spectrum pesticide ‘chlorothalonil, CHT' were selected as the experimental materials. Then, we studied the effects of exogenous SAM on the metabolism of pesticide, reactive oxygen species and glutathione. 【Result】 The results showed that the CHT residues were significantly reduced by exogenous spraying 0.5-2 μmol·L~(-1) SAM. However, no significantly changes could be observed when the concentrations of SAM arrived at 0.5 μmol·L~(-1). Compared with control, the activities of glutathione S-transferase(GST), glutathione reductase(GR) and dehydroascorbate reductase(DHAR) were significantly increased under CHT treatment. And similar results could be obtained with the contents of glutathione(GSH), oxidized glutathione(GSSG) and total glutathione(GSH+GSSG). The GSH/GSSG were induced by CHT firstly, and then decreased. At the same time, superoxide anion(O_2~-) and hydrogen peroxide(H_2O_2) were significantly induced by CHT. However, no significant changes were observed in malondialdehyde(MDA) content. Importantly, exogenous application of 0.5 μmol·L~(-1) SAM had significant effects on improving the activities of GST, GR and DHAR, as well as contents of GSH and GSH+GSSG. Similarly, no significant changes were observed in MDA contents among different treatments. 【Conclusion】 Taken together, exogenous application of 0.5 μmol·L~(-1) SAM could active the key enzymes of GR and DHAR to improve the glutathione cycle and GSH regeneration, which induced the CHT metabolism of tomato fruit depending on GST, O_2~-, and H_2O_2 pathway.
【Objective】 In order to provide basic mechanism for vegetable safe production, this study aimed at revealing the physiological mechanism of S-adenosylmethionine(SAM)-induced pesticide metabolism. 【Method】 The fresh-eating tomato‘QianXi' and broad-spectrum pesticide ‘chlorothalonil, CHT' were selected as the experimental materials. Then, we studied the effects of exogenous SAM on the metabolism of pesticide, reactive oxygen species and glutathione. 【Result】 The results showed that the CHT residues were significantly reduced by exogenous spraying 0.5-2 μmol·L~(-1) SAM. However, no significantly changes could be observed when the concentrations of SAM arrived at 0.5 μmol·L~(-1). Compared with control, the activities of glutathione S-transferase(GST), glutathione reductase(GR) and dehydroascorbate reductase(DHAR) were significantly increased under CHT treatment. And similar results could be obtained with the contents of glutathione(GSH), oxidized glutathione(GSSG) and total glutathione(GSH+GSSG). The GSH/GSSG were induced by CHT firstly, and then decreased. At the same time, superoxide anion(O_2~-) and hydrogen peroxide(H_2O_2) were significantly induced by CHT. However, no significant changes were observed in malondialdehyde(MDA) content. Importantly, exogenous application of 0.5 μmol·L~(-1) SAM had significant effects on improving the activities of GST, GR and DHAR, as well as contents of GSH and GSH+GSSG. Similarly, no significant changes were observed in MDA contents among different treatments. 【Conclusion】 Taken together, exogenous application of 0.5 μmol·L~(-1) SAM could active the key enzymes of GR and DHAR to improve the glutathione cycle and GSH regeneration, which induced the CHT metabolism of tomato fruit depending on GST, O_2~-, and H_2O_2 pathway.
引文
[1]于安芬,车宗贤,王东晖,李瑞琴,白滨,徐瑞,宋政平.百菌清在番茄上的自然消解动态及健康风险研究.西北农业学报,2016,25(10):1508-1514.YU A F,CHE Z X,WANG D H,LI R Q,BAI B,XU R,SONG Z P.Study on natural dynamic degradation of chlorothalonil in tomato and its health risks.Acta Agriculturae Boreali-occidentalis Sinica,2016,25(10):1508-1514.(in Chinese)
[2]ZHOU Y H,XIA X J,YU G B,WANG J T,WU J X,WANG M M,YANG Y X,SHI K,YU Y L,CHEN Z X,GAN J,YU J Q.Brassinosteroids play a critical role in the regulation of pesticide metabolism in crop plants.Scientific Reports,2015,5:9018.
[3]MUNTANE J.Regulation of drug metabolism and transporters.Current Drug Metabolism,2009,10(8):932-945.
[4]SIMINSZKY B.Plant cytochrome P450-mediated herbicide metabolism.Phytochemistry Reviews,2006,5:445-458.
[5]YIN Y L,ZHOU Y,ZHOU Y H,SHI K,ZHOU J,YU Y L,YU J Q,XIA X J.Interplay between mitogen-activated protein kinase and nitric oxide in brassinosteroid-induced pesticide metabolism in Solanum lycopersicum.Journal of Hazardous Materials,2016,316:221-231.
[6]杜丽亚,章钢娅,靳伟.土壤含水量和胡敏酸对有机氯农药降解的影响.土壤学报,2006,43(2):158-162.DU Y L,ZHANG G Y,JIN W.Effects of soil water content and humic acid on degradation of organochlorine pesticides.Acta Pedologica Sinica,2006,43(2):158-162.(in Chinese)
[7]刘鑫,李晓彤,荆鑫,王硕硕,巩彪,魏珉,史庆华.S-腺苷甲硫氨酸对黄瓜断根扦插苗生长及生理代谢的影响.园艺学报,2018,45(8):1513-1522.LIU X,LI X T,JING X,WANG S S,GONG B,WEI M,SHI Q H.Effect of S-adenosylmethionine on growth and physiological metabolism of cucumber cutting seedlings.Acta Horticulturae Sinica,2018,45(8):1513-1522.(in Chinese)
[8]GONG B,LI X,VANDENLANGENBERG K M,WEN D,SUN S S,WEI M,LI Y,YANG F J,SHI Q H,WANG X F.Overexpression of S-adenosyl-L-methionine synthetase increased tomato tolerance to alkali stress through polyamine metabolism.Plant Biotechnology Journal,2014,12(6):694-708.
[9]GONG B,WANG X F,WEI M,YANG F J,LI Y,SHI Q H.Overexpression of S-adenosylmethionine synthetase 1 enhances tomato callus tolerance to alkali stress through polyamine and hydrogen peroxide cross-linked networks.Plant Cell,Tissus and Organ Culture,2016,124:377-391.
[10]CHEN Y W,TAN T W.Enhanced S-adenosylmethionine production by increasing ATP levels in baker's yeast(Saccharomyces cerevisiae).Journal of Agricultural and Food Chemistry,2018,66(20):5200-5209.
[11]何建社,张利,刘千里,朱欣伟,刘兴良,冯秋红,罗达,史作民.岷江干旱河谷区典型灌木对干旱胁迫的生理生化响应.生态学报,2018,38(7):2362-2371.HE J S,ZHANG L,LIU Q L,ZHU X W,LIU X L,FENG Q H,LUOD,SHI Z M.Physiological and biochemical response of typical shrubs to drought stress in the Minjiang River dry valley.Acta Ecologica Sinica,2018,38(7):2362-2371.(in Chinese)
[12]孙莎莎,巩彪,温丹,王秀峰,魏珉,杨凤娟,李岩,史庆华.对羟基苯甲酸胁迫下褪黑素对黄瓜胚根生理生化特性的影响.应用生态学报,2016,27(3):897-903.SUN S S,GONG B,WEN D,WANG X F,WEI M,YANG F J,LI Y,SHI Q H.Effect of exogenous melatonin on physiological and biochemical characteristics of cucumber radicles under p-hydroxybenzoic acid.Chinese Journal of Applied Ecology,2016,27(3):897-903.(in Chinese)
[13]GONG B,LI X,BLOSZIES S,WEN D,SUN S S,WEI M,LI Y,YANG F J,SHI Q H,WANG X F.Sodic alkaline stress mitigation by interaction of nitric oxide and polyamines involves antioxidants and physiological strategies in Solanum lycopersicum.Free Radical Biology and Medicine,2014,71:36-48.
[14]李顺欣,郅军锐,杨广明,岳文波,叶茂.外源茉莉酸诱导的菜豆叶片生化抗性及其对西花蓟马体内保护酶和解毒酶活性的影响.应用生态学报,2017,28(9):2975-2983.LI S X,ZHI J R,YANG G M,YUE W B,YE M.Resistance of bean leaves induced by exogenous jasmonic acid and its effects on activities of protective and detoxification enzymes in Frankliniella occidentalis.Chinese Journal of Applied Ecology,2017,28(9):2975-2983.(in Chinese)
[15]FOYER C H,HALLIWELL B.The presence of glutathione and glutathione reductase in chloroplasts:A proposed role in ascorbic acid metabolism.Planta,1976,133:21-25.
[16]NAKANO Y,ASADA K.Hydrogen peroxide scanvenged by ascorbate-specific peroxidase in spinach chloroplast.Plant and Cell Physiology,1981,22:867-880.
[17]GONG B,WEN D,BLOSZIES S,LI X,WEI M,YANG F J,SHI Q H,WANG X F.Comparative effects of Na Cl and Na HCO3 stresses on respiratory metabolism,antioxidant system,nutritional status,and organic acid metabolism in tomato roots.Acta Physiologiae Plantarum,2014,36:2167-2181.
[18]柴兴苹,张玉秀,谭金娟,冯珊珊,柴团耀.Zn胁迫下小麦S-腺苷甲硫氨酸代谢途径关键基因表达模式分析.植物生理学报,2013,49(4):375-384.CHAI X P,ZHANG Y X,TAN J J,FENG S S,CHAI T Y.Analysis of expression patterns of genes participated in S-adenosylmethionine(SAM)metabolic pathway in wheat under Zn stress.Plant Physiology Journal,2013,49(4):375-384.(in Chinese)
[19]EZAKI B,HIGASHI A,NANBA N,NISHIUCHI T.An S-adenosyl methionine synthetase(SAMS)gene from Andropogon virginicus L.confers aluminum stress tolerance and facilitates epigenetic gene regulation in Arabidopsis thaliana.Frontiers in Plant Science,2016,7:1627.
[20]NELSON D,WERCK-REICHHART D A.P450-centric view of plant evolution.Plant Journal,2011,66(1):194-211.
[21]LV B S,TIAN H Y,ZHANG F,LIU J J,LU S C,BAI M Y,LI C Y,DING Z J.Brassinosteroids regulate root growth by controlling reactive oxygen species homeostasis and dual effect on ethylene synthesis in Arabidopsis.PLoS Genetics,2018,14(1):e1007144.
[22]NOCTOR G,FOYER C H.Intracellular redox compartmentation and ROS-related communication in regulation and signaling.Plant Physiology,2016,171(3):1581-1592.
[23]GUO Z F,TAN J L,ZHUO C L,WANG C Y,XIANG B,WANG Z Y.Abscisic acid,H2O2 and nitric oxide interactions mediated coldinduced S-adenosylmethionine synthetase in Medicago sativa subsp.falcata that confers cold tolerance through up-regulating polyamine oxidation.Plant Biotechnology Journal,2014,12:601-612.
[24]丁顺华,陈珊,卢从明.植物叶绿体谷胱甘肽还原酶的功能研究进展.植物生理学报,2016,52(11):1703-1709.DING S H,CHEN S,LU C M.Research progress on functions of glutathione reductase in chloroplasts of plants.Plant Physiology Journal,2016,52(11):1703-1709.(in Chinese)
[25]谭维嘉,梁革梅,郭予元,王武刚.B.t杀虫剂预处理缓解棉铃虫对化学农药抗性及机理的研究.中国农业科学,1997,30(5):13-19.TAN J W,LIANG G M,GUO Y Y,WANG W G.Study on the mechanism alleviation of cotton bollworm to insecticides by B.t treatment.Scientia Agricultura Sinica,1997,30(5):13-19.(in Chinese)
[26]KWAK M K,LEE M H,PARK S J,SHIN S M,LIU R,KANG S O.Polyamines regulate cell growth and cellular methylglyoxal in high-glucose medium independently of intracellular glutathione.Febs Letters,2016,590(6):739-749.
[27]DIAO Q N,SONG Y J,QI H Y.Exogenous spermidine enhances chilling tolerance of tomato(Solanum lycopersicum L.)seedlings via involvement in polyamines metabolism and physiological parameter levels.Acta Physiologiae Plantarum,2015,37(11):230.
[28]AGRAWAL S B,AGRAWAL M,LEE E H,KRAMER G F,PILLAI P.Changes in polyamine and glutathione contents of a green alga,Chlorogonium elongatum(Dang)France exposed to mercury.Environmental&Experimental Botany,1992,32(2):145-151.
[29]化文平,刘文超,王喆之,李翠芹.干涉丹参SmORA1对植物抗病和丹参酮类次生代谢的影响.中国农业科学,2016,49(3):491-502.HUA W P,LIU W C,WANG Z Z,LI C Q.Effect of RNAi of SmORA1 on disease resistance and tanshinones secondary metabolism in Salvia miltiorrhiza.Scientia Agricultura Sinica,2016,49(3):491-502.(in Chinese)
[30]周琦,郑幸果,何辉煌,康杰,肖海华.植物S-腺苷甲硫氨酸合成酶的新功能展望.生命的化学,2017(4):57-63.ZHOU Q,ZHENG X G,HE H H,KANG J,XIAO H H.Progresses in new functions of plant S-adenosylmethionine synthetase.Chemistry of Life,2017(4):57-63.(in Chinese)
[2]ZHOU Y H,XIA X J,YU G B,WANG J T,WU J X,WANG M M,YANG Y X,SHI K,YU Y L,CHEN Z X,GAN J,YU J Q.Brassinosteroids play a critical role in the regulation of pesticide metabolism in crop plants.Scientific Reports,2015,5:9018.
[3]MUNTANE J.Regulation of drug metabolism and transporters.Current Drug Metabolism,2009,10(8):932-945.
[4]SIMINSZKY B.Plant cytochrome P450-mediated herbicide metabolism.Phytochemistry Reviews,2006,5:445-458.
[5]YIN Y L,ZHOU Y,ZHOU Y H,SHI K,ZHOU J,YU Y L,YU J Q,XIA X J.Interplay between mitogen-activated protein kinase and nitric oxide in brassinosteroid-induced pesticide metabolism in Solanum lycopersicum.Journal of Hazardous Materials,2016,316:221-231.
[6]杜丽亚,章钢娅,靳伟.土壤含水量和胡敏酸对有机氯农药降解的影响.土壤学报,2006,43(2):158-162.DU Y L,ZHANG G Y,JIN W.Effects of soil water content and humic acid on degradation of organochlorine pesticides.Acta Pedologica Sinica,2006,43(2):158-162.(in Chinese)
[7]刘鑫,李晓彤,荆鑫,王硕硕,巩彪,魏珉,史庆华.S-腺苷甲硫氨酸对黄瓜断根扦插苗生长及生理代谢的影响.园艺学报,2018,45(8):1513-1522.LIU X,LI X T,JING X,WANG S S,GONG B,WEI M,SHI Q H.Effect of S-adenosylmethionine on growth and physiological metabolism of cucumber cutting seedlings.Acta Horticulturae Sinica,2018,45(8):1513-1522.(in Chinese)
[8]GONG B,LI X,VANDENLANGENBERG K M,WEN D,SUN S S,WEI M,LI Y,YANG F J,SHI Q H,WANG X F.Overexpression of S-adenosyl-L-methionine synthetase increased tomato tolerance to alkali stress through polyamine metabolism.Plant Biotechnology Journal,2014,12(6):694-708.
[9]GONG B,WANG X F,WEI M,YANG F J,LI Y,SHI Q H.Overexpression of S-adenosylmethionine synthetase 1 enhances tomato callus tolerance to alkali stress through polyamine and hydrogen peroxide cross-linked networks.Plant Cell,Tissus and Organ Culture,2016,124:377-391.
[10]CHEN Y W,TAN T W.Enhanced S-adenosylmethionine production by increasing ATP levels in baker's yeast(Saccharomyces cerevisiae).Journal of Agricultural and Food Chemistry,2018,66(20):5200-5209.
[11]何建社,张利,刘千里,朱欣伟,刘兴良,冯秋红,罗达,史作民.岷江干旱河谷区典型灌木对干旱胁迫的生理生化响应.生态学报,2018,38(7):2362-2371.HE J S,ZHANG L,LIU Q L,ZHU X W,LIU X L,FENG Q H,LUOD,SHI Z M.Physiological and biochemical response of typical shrubs to drought stress in the Minjiang River dry valley.Acta Ecologica Sinica,2018,38(7):2362-2371.(in Chinese)
[12]孙莎莎,巩彪,温丹,王秀峰,魏珉,杨凤娟,李岩,史庆华.对羟基苯甲酸胁迫下褪黑素对黄瓜胚根生理生化特性的影响.应用生态学报,2016,27(3):897-903.SUN S S,GONG B,WEN D,WANG X F,WEI M,YANG F J,LI Y,SHI Q H.Effect of exogenous melatonin on physiological and biochemical characteristics of cucumber radicles under p-hydroxybenzoic acid.Chinese Journal of Applied Ecology,2016,27(3):897-903.(in Chinese)
[13]GONG B,LI X,BLOSZIES S,WEN D,SUN S S,WEI M,LI Y,YANG F J,SHI Q H,WANG X F.Sodic alkaline stress mitigation by interaction of nitric oxide and polyamines involves antioxidants and physiological strategies in Solanum lycopersicum.Free Radical Biology and Medicine,2014,71:36-48.
[14]李顺欣,郅军锐,杨广明,岳文波,叶茂.外源茉莉酸诱导的菜豆叶片生化抗性及其对西花蓟马体内保护酶和解毒酶活性的影响.应用生态学报,2017,28(9):2975-2983.LI S X,ZHI J R,YANG G M,YUE W B,YE M.Resistance of bean leaves induced by exogenous jasmonic acid and its effects on activities of protective and detoxification enzymes in Frankliniella occidentalis.Chinese Journal of Applied Ecology,2017,28(9):2975-2983.(in Chinese)
[15]FOYER C H,HALLIWELL B.The presence of glutathione and glutathione reductase in chloroplasts:A proposed role in ascorbic acid metabolism.Planta,1976,133:21-25.
[16]NAKANO Y,ASADA K.Hydrogen peroxide scanvenged by ascorbate-specific peroxidase in spinach chloroplast.Plant and Cell Physiology,1981,22:867-880.
[17]GONG B,WEN D,BLOSZIES S,LI X,WEI M,YANG F J,SHI Q H,WANG X F.Comparative effects of Na Cl and Na HCO3 stresses on respiratory metabolism,antioxidant system,nutritional status,and organic acid metabolism in tomato roots.Acta Physiologiae Plantarum,2014,36:2167-2181.
[18]柴兴苹,张玉秀,谭金娟,冯珊珊,柴团耀.Zn胁迫下小麦S-腺苷甲硫氨酸代谢途径关键基因表达模式分析.植物生理学报,2013,49(4):375-384.CHAI X P,ZHANG Y X,TAN J J,FENG S S,CHAI T Y.Analysis of expression patterns of genes participated in S-adenosylmethionine(SAM)metabolic pathway in wheat under Zn stress.Plant Physiology Journal,2013,49(4):375-384.(in Chinese)
[19]EZAKI B,HIGASHI A,NANBA N,NISHIUCHI T.An S-adenosyl methionine synthetase(SAMS)gene from Andropogon virginicus L.confers aluminum stress tolerance and facilitates epigenetic gene regulation in Arabidopsis thaliana.Frontiers in Plant Science,2016,7:1627.
[20]NELSON D,WERCK-REICHHART D A.P450-centric view of plant evolution.Plant Journal,2011,66(1):194-211.
[21]LV B S,TIAN H Y,ZHANG F,LIU J J,LU S C,BAI M Y,LI C Y,DING Z J.Brassinosteroids regulate root growth by controlling reactive oxygen species homeostasis and dual effect on ethylene synthesis in Arabidopsis.PLoS Genetics,2018,14(1):e1007144.
[22]NOCTOR G,FOYER C H.Intracellular redox compartmentation and ROS-related communication in regulation and signaling.Plant Physiology,2016,171(3):1581-1592.
[23]GUO Z F,TAN J L,ZHUO C L,WANG C Y,XIANG B,WANG Z Y.Abscisic acid,H2O2 and nitric oxide interactions mediated coldinduced S-adenosylmethionine synthetase in Medicago sativa subsp.falcata that confers cold tolerance through up-regulating polyamine oxidation.Plant Biotechnology Journal,2014,12:601-612.
[24]丁顺华,陈珊,卢从明.植物叶绿体谷胱甘肽还原酶的功能研究进展.植物生理学报,2016,52(11):1703-1709.DING S H,CHEN S,LU C M.Research progress on functions of glutathione reductase in chloroplasts of plants.Plant Physiology Journal,2016,52(11):1703-1709.(in Chinese)
[25]谭维嘉,梁革梅,郭予元,王武刚.B.t杀虫剂预处理缓解棉铃虫对化学农药抗性及机理的研究.中国农业科学,1997,30(5):13-19.TAN J W,LIANG G M,GUO Y Y,WANG W G.Study on the mechanism alleviation of cotton bollworm to insecticides by B.t treatment.Scientia Agricultura Sinica,1997,30(5):13-19.(in Chinese)
[26]KWAK M K,LEE M H,PARK S J,SHIN S M,LIU R,KANG S O.Polyamines regulate cell growth and cellular methylglyoxal in high-glucose medium independently of intracellular glutathione.Febs Letters,2016,590(6):739-749.
[27]DIAO Q N,SONG Y J,QI H Y.Exogenous spermidine enhances chilling tolerance of tomato(Solanum lycopersicum L.)seedlings via involvement in polyamines metabolism and physiological parameter levels.Acta Physiologiae Plantarum,2015,37(11):230.
[28]AGRAWAL S B,AGRAWAL M,LEE E H,KRAMER G F,PILLAI P.Changes in polyamine and glutathione contents of a green alga,Chlorogonium elongatum(Dang)France exposed to mercury.Environmental&Experimental Botany,1992,32(2):145-151.
[29]化文平,刘文超,王喆之,李翠芹.干涉丹参SmORA1对植物抗病和丹参酮类次生代谢的影响.中国农业科学,2016,49(3):491-502.HUA W P,LIU W C,WANG Z Z,LI C Q.Effect of RNAi of SmORA1 on disease resistance and tanshinones secondary metabolism in Salvia miltiorrhiza.Scientia Agricultura Sinica,2016,49(3):491-502.(in Chinese)
[30]周琦,郑幸果,何辉煌,康杰,肖海华.植物S-腺苷甲硫氨酸合成酶的新功能展望.生命的化学,2017(4):57-63.ZHOU Q,ZHENG X G,HE H H,KANG J,XIAO H H.Progresses in new functions of plant S-adenosylmethionine synthetase.Chemistry of Life,2017(4):57-63.(in Chinese)