低温胁迫下番茄幼苗根穗互作对其抗坏血酸—谷胱甘肽循环的影响
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摘要
为探讨番茄根系与接穗及其互作效应影响嫁接苗耐冷性的效应大小与生理机制,以耐冷性强的番茄‘060112’(R)和冷敏感番茄‘060911’(S)为试材,采用靠接法进行嫁接,组成双根双穗嫁接苗RS/RS,待嫁接苗成活后通过断根或断穗,形成R/R、R/RS、R/S、S/R、S/RS和S/S共6个根/穗组合处理,置于光强22 klx、光周期12 h/12 h光照培养箱,测定昼夜温度10℃/3℃低温胁迫9 d及25℃/15℃常温恢复3 d时嫁接苗叶片和根系中活性氧水平及As A-GSH循环中关键酶和抗氧化物质的变化。结果表明,低温胁迫显著提高了番茄嫁接苗叶片和根系中H_2O_2含量及超氧阴离子产生速率,但耐冷性强的R根系嫁接苗活性氧水平显著低于冷敏感的S根系嫁接苗,虽然R接穗嫁接苗活性氧水平亦显著低于S接穗嫁接苗,但降幅不及根系嫁接苗的大,表明根系在增强嫁接苗耐冷性中的作用大于接穗。低温胁迫下,各处理嫁接苗叶片和根系As A-GSH循环中关键酶(APX、MDHAR、DHAR、GR)活性和抗氧化物质(As A、DHA、GSH、GSSG)的含量均显著增加,且除DHA和GSSG外,均以R根系或R接穗嫁接苗较高,S根系或S接穗嫁接苗较低,RS根系或RS接穗嫁接苗居中,表明低温胁迫下耐冷性强的R根系或接穗嫁接苗的抗氧化能力显著高于S根系或接穗嫁接苗。具RS双接穗的嫁接苗中又以RS-R叶片的抗氧化能力显著高于RS-S叶片,且互作效应的P值小于0.01(或0.05),表明根、穗对嫁接苗耐冷性存在显著的互作效应。
In order to investigate the effect of tomato rootstock,scion and their potential interaction on chilling tolerance,the approach grafting between chilling-sensitive tomato cultivar‘060911'(S)and chilling-resistant tomato cultivar‘060112'(R)was proceeded to generate grafted seedlings RS/RS with double rootstocks and scions. By selectively detaching one rootstock or scion from RS/RS,newly formed seedlings of six combinations with different rootstock and scion,i.e.,R/R,R/RS,R/S,S/R,S/RS,S/S were raised in the growth chamber with light intensity of 22 klx and photoperiod of 12 h/12 h. Seedlings were treated with low temperature at 10 ℃ in the day and 3 ℃ at night for 9 days and recovered at 25 ℃ in the day and 15 ℃ at night for 3 days. The level of reactive oxygen species(ROS),and the key enzymes and antioxidants in AsA-GSH cycle in leaves and roots of grafted seedlings were measured after the chilling treatment and recovery. The content of hydrogen peroxide and production rate of superoxide anion in leaves and roots of grafted tomato seedlings were significantly increased under low temperature stress. The ROS level in grafted seedlings with R rootstock or R scion was significantly lower than that with S rootstock or S scion,respectively. The decrease of ROS level between R and S rootstock was more significant than that between R and S scion,indicating that rootstock conferred more chilling resistance than scion. Low temperature stress increased the accumulation of AsA,DHA,GSH and GSSG,and enhanced the activities of APX,MDHAR,DHAR and GR. Moreover,enzyme activities and the content of antioxidants except for DHA and GSSG were highest in grafted seedlings with R rootstock or R scion,lowest in those with S rootstock or scion,and intermediate in those with RS rootstock or scion,indicating that the ability of anti-oxidation in grafted seedlings with R rootstock or R scion under cold stress was significantly higher than that in grafted seedlings with S rootstock or S scion. In addition,the ability of anti-oxidation in leaves of RS-R grafted seedling was significantly higher than that in RS-S grafted seedlings. Statistical analysis showed that probability(P)values of interaction between rootstock and scion were less than 0.01(or 0.05),indicating that rootstock and scion posses significant interaction effects on the chilling tolerance of grafted tomato seedlings.
In order to investigate the effect of tomato rootstock,scion and their potential interaction on chilling tolerance,the approach grafting between chilling-sensitive tomato cultivar‘060911'(S)and chilling-resistant tomato cultivar‘060112'(R)was proceeded to generate grafted seedlings RS/RS with double rootstocks and scions. By selectively detaching one rootstock or scion from RS/RS,newly formed seedlings of six combinations with different rootstock and scion,i.e.,R/R,R/RS,R/S,S/R,S/RS,S/S were raised in the growth chamber with light intensity of 22 klx and photoperiod of 12 h/12 h. Seedlings were treated with low temperature at 10 ℃ in the day and 3 ℃ at night for 9 days and recovered at 25 ℃ in the day and 15 ℃ at night for 3 days. The level of reactive oxygen species(ROS),and the key enzymes and antioxidants in AsA-GSH cycle in leaves and roots of grafted seedlings were measured after the chilling treatment and recovery. The content of hydrogen peroxide and production rate of superoxide anion in leaves and roots of grafted tomato seedlings were significantly increased under low temperature stress. The ROS level in grafted seedlings with R rootstock or R scion was significantly lower than that with S rootstock or S scion,respectively. The decrease of ROS level between R and S rootstock was more significant than that between R and S scion,indicating that rootstock conferred more chilling resistance than scion. Low temperature stress increased the accumulation of AsA,DHA,GSH and GSSG,and enhanced the activities of APX,MDHAR,DHAR and GR. Moreover,enzyme activities and the content of antioxidants except for DHA and GSSG were highest in grafted seedlings with R rootstock or R scion,lowest in those with S rootstock or scion,and intermediate in those with RS rootstock or scion,indicating that the ability of anti-oxidation in grafted seedlings with R rootstock or R scion under cold stress was significantly higher than that in grafted seedlings with S rootstock or S scion. In addition,the ability of anti-oxidation in leaves of RS-R grafted seedling was significantly higher than that in RS-S grafted seedlings. Statistical analysis showed that probability(P)values of interaction between rootstock and scion were less than 0.01(or 0.05),indicating that rootstock and scion posses significant interaction effects on the chilling tolerance of grafted tomato seedlings.
引文
Ahmad P,Nabi G,Ashraf M.2011.Cadmium-induced oxidative damage in mustard[Brassica juncea(L.)Czern.&Coss.]plants can be alleviated by salicylic acid.South African Journal of Botany,77(1):36-44.
Airaki M,Leterrier M,Mateos R M,Valderrama R,Chaki M,Barroso J B,LA Del RI′O,Palma J M,Corpas F J.2012.Metabolism of reactive oxygen species and reactive nitrogen species in pepper(Capsicum annuum L.)plants under low temperature stress.Plant Cell Environ,35(2):281-295.
Anderson J V,Chevone B I,Hess J L.1992.Seasonal variation in the antioxidant system of eastern white pine needles.Plant physiology,98(2):501-508.
Chen J L,Wu X X,Yao X F,Zhu Z W,Xu S,Zha D S.2016.Exogenous 6-benzylaminopurine confers tolerance to low temperature by amelioration of oxidative damage in eggplant(Solanum melongena L.)seedlings.Brazilian Journal of Botany,39(2):409-416.
Chen S,Yin C,Strasser R J,Govindjee,Yang C,Qiang S.2012.Reactive oxygen species from chloroplasts contribute to 3-acetyl-5-isopropyltetramic acid-induced leaf necrosis of Arabidopsis thaliana.Plant Physiol Bioch,52(1):38-51.
Chen Z,Gallie D R.2006.Dehydroascorbate reductase affects leaf growth,development,and function.Plant Physiology,142(2):775-787.
Farooq M A,Islam F,Yang C,Nawaz A,Athar H R,Gill R A,Ali B,Song W J,Zhou W J.2017.Methyl jasmonate alleviates arsenic-induced oxidative damage and modulates the ascorbate-glutathione cycle in oilseed rape roots.Plant Growth Regulation.Springer:1-14.
Franck T,Kevers C,Gaspar T.1995.Protective enzymatic systems against activated oxygen species compared in normal and vitrified shoots of Prunus avium L.raised in vitro.Plant growth regulation,16(3):253-256.
Gao Jun-jie,Qin Ai-guo,Yu Xian-chang.2009.Effects of low-temperature stress on the ascorbate-glutathione cycle in the grafted cucumber leaves.Acta Horticulturae Sinica,36(2):215-220.(in Chinese)高俊杰,秦爱国,于贤昌.2009.低温胁迫对嫁接黄瓜叶片抗坏血酸-谷胱甘肽循环的影响.园艺学报,36(2):215-220.
Gay C,Gebicki J M.2000.A critical evaluation of the effect of sorbitol on the ferric-xylenol orange hydroperoxide assay.Analytical Biochemistry,284(2):217-220.
Gill S S,Tuteja N.2010.Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants.Plant Physiol Biochem,48(12):909-930.
Han Min,Cao Bi-li,Liu Shu-sen,Xu Kun.2018a.Effects of rootstock and scion interaction on chilling tolerance of grafted tomato seedlings.Acta Horticulturae Sinica,45(2):279-288.(in Chinese)韩敏,曹逼力,刘树森,徐坤.2018a.番茄嫁接苗根穗互作对其耐冷性的影响.园艺学报,45(2):279-288.
Han Min,Cao Bi-li,Liu Shu-sen,Xu Kun.2018b.Effect of rootstock and scion interaction on photosynthesis and nitrogen metabolism of grafted tomato seedlings leaves under low temperature stress.Acta Horticulturae Sinica,45(5):897-907.(in Chinese)韩敏,曹逼力,刘树森,徐坤.2018b.低温胁迫下番茄嫁接苗根穗互作对叶片光合作用及氮代谢的影响.园艺学报,45(5):897-907.
Hasanuzzaman M,Nahar K,Anee T I,Fujita M.2017.Glutathione in plants:biosynthesis and physiological role in environmental stress tolerance.Physiology&Molecular Biology of Plants An International Journal of Functional Plant Biology,23(2):249-268.
Law M Y,Charles S A,Halliwell B.1983.Glutathione and ascorbic acid in spinach(Spinacia oleracea)chloroplasts.The effect of hydrogen peroxide and of Paraquat.Biochemical Journal,210(3):899-903.
Li L J,Lu X C,Ma H Y,Lyu D G.2017a.Jasmonic acid regulates the ascorbate-glutathione cycle in Malus baccata Borkh.roots under low root-zone temperature.Acta Physiologiae Plantarum,39(174):1-11.
Li Y,Fan Y,Ma Y,Zhang Z,Yue H B,Wang L J,Li J,Jiao Y.2017b.Effects of exogenousγ-aminobutyric acid(GABA)on photosynthesis and antioxidant system in pepper(Capsicum annuum L.)seedlings under low light stress.Journal of Plant Growth Regulation,36(2):1-14.
Li Zhong-guang,Gong Ming.2005.Improvement of measurement method for superoxide anion radical in plant.Acta Botanica Yunnanica,27(2):211-216.(in Chinese)李忠光,龚明.2005.植物中超氧阴离子自由基测定方法的改进.云南植物研究,27(2):211-216.
Liu Y J,Zhao Z G,Si J,Di C X,Han J,An L Z.2009.Brassinosteroids alleviate chilling-induced oxidative damage by enhancing antioxidant defense system in suspension cultured cells of Chorispora bungeana.Plant Growth Regul,59(3):207-214.
Mir B A,Mir S A,Khazir J,Tonfack L B,Cowan D A,Vyasa D,Koula S.2015.Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera(L)Dunal.Industrial Crops&Products,74:1008-1016.
Nahar K,Hasanuzzaman M,Alam M M,Fujita M.2015.Exogenous spermidine alleviates low temperature injury in mung bean(Vigna radiata L.)seedlings by modulating ascorbate-glutathione and glyoxalase pathway.International Journal of Molecular Sciences,16(12):30117-30132.
Poudyal D,Akash M,Khatri L,Shrestha D S,Uptmoor R.2017.Solanum habrochaites introgression line grafted as rootstock in cultivated tomato maintains growth and improves yield under cold and drought stresses.Journal of Crop Improvement,31(4):1-19.
Pukacka S,Ratajczak E.2006.Antioxidative response of ascorbate-glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds.Journal of Plant Physiology,163(12):1259-1266.
Venema J H,Dijk B E,Bax J M,Hasselt P R V,Elzenga J T M.2008.Grafting tomato(Solanum lycopersicum)onto the rootstock of a high-altitude accession of Solanum habrochaites improves suboptimal-temperature tolerance.Environmental and Experimental Botany,63(1):359-367.
Wu X X,He J,Ding H D,Zhu Z W,Chen J L,Xu X,Zha D S.2015.Modulation of zinc-induced oxidative damage in Solanum melongena,by 6-benzylaminopurine involves ascorbate-glutathione cycle metabolism.Environmental&Experimental Botany,116:1-11.
Zhang Y P,Xu S,Yang S J,Chen Y Y.2017.Melatonin alleviates cold-induced oxidative damage by regulation of ascorbateaglutathione and proline metabolism in melon seedlings(Cucumis melo L.).Journal of Pomology&Horticultural Science,92(3):313-324.
Zhao H,Lin Y,Wang Y,Xu H W.2016.Melatonin increases the chilling tolerance of chloroplast in cucumber seedlings by regulating photosynthetic electron flux and the ascorbate-glutathione cycle.Frontiers in Plant Science,7:1-10.
Airaki M,Leterrier M,Mateos R M,Valderrama R,Chaki M,Barroso J B,LA Del RI′O,Palma J M,Corpas F J.2012.Metabolism of reactive oxygen species and reactive nitrogen species in pepper(Capsicum annuum L.)plants under low temperature stress.Plant Cell Environ,35(2):281-295.
Anderson J V,Chevone B I,Hess J L.1992.Seasonal variation in the antioxidant system of eastern white pine needles.Plant physiology,98(2):501-508.
Chen J L,Wu X X,Yao X F,Zhu Z W,Xu S,Zha D S.2016.Exogenous 6-benzylaminopurine confers tolerance to low temperature by amelioration of oxidative damage in eggplant(Solanum melongena L.)seedlings.Brazilian Journal of Botany,39(2):409-416.
Chen S,Yin C,Strasser R J,Govindjee,Yang C,Qiang S.2012.Reactive oxygen species from chloroplasts contribute to 3-acetyl-5-isopropyltetramic acid-induced leaf necrosis of Arabidopsis thaliana.Plant Physiol Bioch,52(1):38-51.
Chen Z,Gallie D R.2006.Dehydroascorbate reductase affects leaf growth,development,and function.Plant Physiology,142(2):775-787.
Farooq M A,Islam F,Yang C,Nawaz A,Athar H R,Gill R A,Ali B,Song W J,Zhou W J.2017.Methyl jasmonate alleviates arsenic-induced oxidative damage and modulates the ascorbate-glutathione cycle in oilseed rape roots.Plant Growth Regulation.Springer:1-14.
Franck T,Kevers C,Gaspar T.1995.Protective enzymatic systems against activated oxygen species compared in normal and vitrified shoots of Prunus avium L.raised in vitro.Plant growth regulation,16(3):253-256.
Gao Jun-jie,Qin Ai-guo,Yu Xian-chang.2009.Effects of low-temperature stress on the ascorbate-glutathione cycle in the grafted cucumber leaves.Acta Horticulturae Sinica,36(2):215-220.(in Chinese)高俊杰,秦爱国,于贤昌.2009.低温胁迫对嫁接黄瓜叶片抗坏血酸-谷胱甘肽循环的影响.园艺学报,36(2):215-220.
Gay C,Gebicki J M.2000.A critical evaluation of the effect of sorbitol on the ferric-xylenol orange hydroperoxide assay.Analytical Biochemistry,284(2):217-220.
Gill S S,Tuteja N.2010.Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants.Plant Physiol Biochem,48(12):909-930.
Han Min,Cao Bi-li,Liu Shu-sen,Xu Kun.2018a.Effects of rootstock and scion interaction on chilling tolerance of grafted tomato seedlings.Acta Horticulturae Sinica,45(2):279-288.(in Chinese)韩敏,曹逼力,刘树森,徐坤.2018a.番茄嫁接苗根穗互作对其耐冷性的影响.园艺学报,45(2):279-288.
Han Min,Cao Bi-li,Liu Shu-sen,Xu Kun.2018b.Effect of rootstock and scion interaction on photosynthesis and nitrogen metabolism of grafted tomato seedlings leaves under low temperature stress.Acta Horticulturae Sinica,45(5):897-907.(in Chinese)韩敏,曹逼力,刘树森,徐坤.2018b.低温胁迫下番茄嫁接苗根穗互作对叶片光合作用及氮代谢的影响.园艺学报,45(5):897-907.
Hasanuzzaman M,Nahar K,Anee T I,Fujita M.2017.Glutathione in plants:biosynthesis and physiological role in environmental stress tolerance.Physiology&Molecular Biology of Plants An International Journal of Functional Plant Biology,23(2):249-268.
Law M Y,Charles S A,Halliwell B.1983.Glutathione and ascorbic acid in spinach(Spinacia oleracea)chloroplasts.The effect of hydrogen peroxide and of Paraquat.Biochemical Journal,210(3):899-903.
Li L J,Lu X C,Ma H Y,Lyu D G.2017a.Jasmonic acid regulates the ascorbate-glutathione cycle in Malus baccata Borkh.roots under low root-zone temperature.Acta Physiologiae Plantarum,39(174):1-11.
Li Y,Fan Y,Ma Y,Zhang Z,Yue H B,Wang L J,Li J,Jiao Y.2017b.Effects of exogenousγ-aminobutyric acid(GABA)on photosynthesis and antioxidant system in pepper(Capsicum annuum L.)seedlings under low light stress.Journal of Plant Growth Regulation,36(2):1-14.
Li Zhong-guang,Gong Ming.2005.Improvement of measurement method for superoxide anion radical in plant.Acta Botanica Yunnanica,27(2):211-216.(in Chinese)李忠光,龚明.2005.植物中超氧阴离子自由基测定方法的改进.云南植物研究,27(2):211-216.
Liu Y J,Zhao Z G,Si J,Di C X,Han J,An L Z.2009.Brassinosteroids alleviate chilling-induced oxidative damage by enhancing antioxidant defense system in suspension cultured cells of Chorispora bungeana.Plant Growth Regul,59(3):207-214.
Mir B A,Mir S A,Khazir J,Tonfack L B,Cowan D A,Vyasa D,Koula S.2015.Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera(L)Dunal.Industrial Crops&Products,74:1008-1016.
Nahar K,Hasanuzzaman M,Alam M M,Fujita M.2015.Exogenous spermidine alleviates low temperature injury in mung bean(Vigna radiata L.)seedlings by modulating ascorbate-glutathione and glyoxalase pathway.International Journal of Molecular Sciences,16(12):30117-30132.
Poudyal D,Akash M,Khatri L,Shrestha D S,Uptmoor R.2017.Solanum habrochaites introgression line grafted as rootstock in cultivated tomato maintains growth and improves yield under cold and drought stresses.Journal of Crop Improvement,31(4):1-19.
Pukacka S,Ratajczak E.2006.Antioxidative response of ascorbate-glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds.Journal of Plant Physiology,163(12):1259-1266.
Venema J H,Dijk B E,Bax J M,Hasselt P R V,Elzenga J T M.2008.Grafting tomato(Solanum lycopersicum)onto the rootstock of a high-altitude accession of Solanum habrochaites improves suboptimal-temperature tolerance.Environmental and Experimental Botany,63(1):359-367.
Wu X X,He J,Ding H D,Zhu Z W,Chen J L,Xu X,Zha D S.2015.Modulation of zinc-induced oxidative damage in Solanum melongena,by 6-benzylaminopurine involves ascorbate-glutathione cycle metabolism.Environmental&Experimental Botany,116:1-11.
Zhang Y P,Xu S,Yang S J,Chen Y Y.2017.Melatonin alleviates cold-induced oxidative damage by regulation of ascorbateaglutathione and proline metabolism in melon seedlings(Cucumis melo L.).Journal of Pomology&Horticultural Science,92(3):313-324.
Zhao H,Lin Y,Wang Y,Xu H W.2016.Melatonin increases the chilling tolerance of chloroplast in cucumber seedlings by regulating photosynthetic electron flux and the ascorbate-glutathione cycle.Frontiers in Plant Science,7:1-10.