外源γ-氨基丁酸对Ca(NO_3)_2胁迫下甜瓜幼苗NO_3~--N同化的影响
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摘要
以盐敏感型甜瓜品种‘一品天下208'为试材用80mmol·L~(-1)Ca(NO_3)_2模拟设施土壤盐渍化采用深液流水培,研究外源γ-氨基丁酸(GABA)对Ca(NO_3)_2胁迫下甜瓜幼苗硝态氮(NO_3~--N)同化的影响.结果表明:Ca(NO_3)_2胁迫显著降低了甜瓜幼苗体内硝酸还原酶(NR)、谷氨酰胺合酶(GS)和谷氨酸合酶(GOGAT)活性,增强了谷氨酸脱氢酶(GDH)、谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性,导致铵态氮(NH_4~+-N)和游离氨基酸含量增加,NO_3~--N和可溶性蛋白质含量下降,植株生长和光合作用受到严重抑制.Ca(NO_3)_2胁迫下,外源喷施GABA有效促进了甜瓜根系对NO_3~--N的吸收及其向地上部的转运,并通过增强NR、GS和GOGAT活性提高了甜瓜幼苗对NH_4~+的同化力;通过抑制GDH脱氨作用减少了甜瓜幼苗体内NH_4~+的释放量,从而缓解了盐诱导产生的NH_4~+-N积累所造成的氨毒害作用;外源喷施GABA也能调节甜瓜组织中氨基酸代谢途径,促进蛋白质的合成.表明外源GABA能增强甜瓜幼苗对NO_3~--N的同化能力,调控氨基酸代谢,进而有效缓解Ca(NO_3)_2胁迫对甜瓜幼苗的盐伤害作用.
The effect of exogenous γ-aminobutyric acid(GABA) on NO_3~--N assimilation in muskmelon under Ca(NO_3)_2 stress was investigated in ' Yipintianxia 208',a salt-sensitive melon variety cultured under deep flow hydroponics which simulated soil salinization.The results showed that under Ca(NO_3)_2 stress,the activities of nitrate reductase(NR),glutamate synthetase(GS) and glutamate amino transferase(GOGAT) in muskmelon seedlings were significantly reduced,while the activities of glutamate dehydrogenase(GDH),glutamate oxaloacetate transaminase(GOT) and glutamate pyruvate aminotransferase(GPT) were enhanced,leading to increased contents of NH_4~+-N and total amino acids,and decreased contents NO_3~--N and soluble protein in muskmelon,which further severely inhibited plant growth and photosynthesis of muskmelon seedlings.Exogenous GABA effectively improved the absorption of NO_3~--N in muskmelon roots and its transportation from root to shoot under Ca(NO_3)_2 stress,and improved NH_4~+-N assimilation by enhancing NR,GS and GOGAT activities in muskmelon seedlings.Exogenous GABA also reduced NH_4 release by limiting GDH deamination,thus further alleviated the toxication of NH_4~+-N induced by Ca(NO_3)_2 stress.In addition,foliage spraying of GABA could regulate amino acids metabolic pathways and promote protein synthesis.The results suggested that exogenous GABA could improve NO_3~--N assimilation and regulate amino acids metabolism to alleviate Ca(NO_3)_2 stress damage in muskmelon seedlings.
The effect of exogenous γ-aminobutyric acid(GABA) on NO_3~--N assimilation in muskmelon under Ca(NO_3)_2 stress was investigated in ' Yipintianxia 208',a salt-sensitive melon variety cultured under deep flow hydroponics which simulated soil salinization.The results showed that under Ca(NO_3)_2 stress,the activities of nitrate reductase(NR),glutamate synthetase(GS) and glutamate amino transferase(GOGAT) in muskmelon seedlings were significantly reduced,while the activities of glutamate dehydrogenase(GDH),glutamate oxaloacetate transaminase(GOT) and glutamate pyruvate aminotransferase(GPT) were enhanced,leading to increased contents of NH_4~+-N and total amino acids,and decreased contents NO_3~--N and soluble protein in muskmelon,which further severely inhibited plant growth and photosynthesis of muskmelon seedlings.Exogenous GABA effectively improved the absorption of NO_3~--N in muskmelon roots and its transportation from root to shoot under Ca(NO_3)_2 stress,and improved NH_4~+-N assimilation by enhancing NR,GS and GOGAT activities in muskmelon seedlings.Exogenous GABA also reduced NH_4 release by limiting GDH deamination,thus further alleviated the toxication of NH_4~+-N induced by Ca(NO_3)_2 stress.In addition,foliage spraying of GABA could regulate amino acids metabolic pathways and promote protein synthesis.The results suggested that exogenous GABA could improve NO_3~--N assimilation and regulate amino acids metabolism to alleviate Ca(NO_3)_2 stress damage in muskmelon seedlings.
引文
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[6]Zhou W-H(周万海),Shi S-L(师尚礼),Kou J-T(寇江涛).Effects of exogenous nitric oxide on the growth and nitrogen metabolism of alfalfa seedlings under salt stress.Chinese Journal of Applied Ecology(应用生态学报),2012,23(11):3003-3008(in Chinese)
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[9]Xiang L-X(向丽霞),Hu L-P(胡立盼),Hu X-H(胡晓辉),et al.Response of reactive oxygen metabolism in melon chloroplasts to short-term salinity-alkalinity stress regulated by exogenousγ-aminobutyric acid.Chinese Journal of Applied Ecology(应用生态学报),2015,26(12):3746-3752(in Chinese)
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[12]Bouche N,Lacombe B,Fromm H.GABA signaling:A conserved and ubiquitous mechanism.Trends in Cell Biology,2003,13:607-610
[13]Fas P,Niogret MF,Montes E,et al.Transcriptional profiling of genes encoding GABA-transaminases in Brassica napus reveals their regulation by water deficit.Environmental and Experimental Botany,2015,116:20-31
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[16]Xu Z-R(徐志然),Zhang L(张丽),Zhang Z(张智),et al.Effects of GABA on tolerance of muskmelon seedlings against Ca(NO3)2stress.Journal of Northwest A&F University(Natural Science)(西北农林科技大学学报:自然科学版),2015,43(3):125-131(in Chinese)
[17]Wu G-P(魏国平),Zhu Y-L(朱月林),Liu Z-L(刘正鲁),et al.Effects of calcium nitrate stress on ascorbate-glutathione cycle metabolism in leaves of hydroponically-grown grafted eggplant seedlings.Chinese Journal of Plant Ecology(植物生态学报),2008,32(5):1023-1030(in Chinese)
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[19]Tang Z-C(汤章城).Modern Plant Physiology Laboratory Manual.Beijing:Science Press,1999(in Chinese)
[20]Singh RD,Srivastava HS.Increase in glutamate synthase(NADH)activity in maize seedling in response to nitrate and ammonium nitrogen.Plant Physiology,1986,66:413-416
[21]Kanamori T,Konishi S,Takahashi E.Inducible formation of glutamate dehydrogenase in rice plant roots by the addition of ammonia to the media.Plant Physiology,1972,26:1-6
[22]Bradford MM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry,1976,72:248-254
[23]Zhang GJ,Bown AW.The rapid determination ofγ-aminobutyric acid.Phytochemistry,1997,44:1007-1009
[24]Tong H(童辉),Zhang Z-X(张振兴),Li B(李斌),et al.Effects of iso-osmotic Ca(NO3)2and Na Cl stress on chloroplast ultrastructure and photosynthesis in cucumber leaves.China Vegetables(中国蔬菜),2012(18):160-165(in Chinese)
[25]Yuan LY,Shu S,Sun J.Effects of 24-epibrassinolide on the photosynthetic characteristics,antioxidant system,and chloroplast ultrastructure in Cucumis sativus L.under Ca(NO3)2stress.Photosynthesis Research,2012,112:205-214
[26]Huang J(黄娟),Li X-F(李兴发),Huang S(黄山),et al.Effects of GABA soaking on cucumber seed germination and seedling growth under different temperature stress.Journal of Changjiang Vegetables(长江蔬菜),2014(12):30-35(in Chinese)
[27]Gao H-B(高洪波),Zhang T-J(章铁军),Lv G-Y(吕桂云),et al.Effects of exogenousγ-aminobutyric acid on growth and reactive oxygen species metabolism of cucumber seedlings under Na Cl stress.Acta Botanica Boreali-Occidentalia Sinica(西北植物学报),2007,27(10):2046-2051(in Chinese)
[28]Gangwar S,Singh VP.Indole acetic acid differently changes growth and nitrogen metabolism in Pisum sativum L.seedlings under chromium(Vl)phytotoxicity:Implication of oxidative stress.Scientia Horticulturae,2011,129:321-32
[29]Zhang Y(张毅),Shi Y(石玉),Hu X-H(胡晓辉),et al.Effects of exogenous spermidine on nitrogen metabolism and contents of main mineral elements in tomato seedlings under salinity-alkalinity mixed stress.Chinese Journal of Applied Ecology(应用生态学报),2013,24(5):1401-1408(in Chinese)
[30]Song S-L(宋锁玲),Li J-R(李敬蕊),Gao H-B(高洪波),et al.Effects of exogenousγ-aminobutyric acid on inorganic nitrogen metabolism and mineral elements contents of melon seedling under hypoxia stress.Acta Horticulturae Sinica(园艺学报),2012,39(4):695-704(in Chinese)
[31]Wang XP,Geng SJ,Ri YJ,et al.Physiological responses and adaptive strategies of tomato plants to salt and alkali stresses.Scientia Horticulturae,2011,130:248-255
[32]Li B(李冰),Zhang Z-G(张照贵),Wang J-J(王佳佳),et al.Cloning and functional marker of GDH1gene in wheat.Shandong Agricultural Sciences(山东农业科学),2014,46(10):6-11(in Chinese)
[33]Liang CG,Chen LP,Wang Y,et al.High temperature at grain-filling stage affects nitrogen metabolism enzyme activities in grains and grain nutritional quality in rice.Rice Science,2011,18:210-216
[34]Ferreira-Silva SL,Silva EN,Carvalho FEL,et al.Physiological alterations modulated by rootstock and scion combination in cashew under salinity.Hortscience,2010,127:39-45
[35]Kang Y-Y(康云艳),Guo S-R(郭世荣),Duan J-J(段九菊).Effects of root zone hypoxia on respiratory metabolism of cucumber seedlings roots.Chinese Journal of Applied Ecology(应用生态学报),2008,19(3):583-587(in Chinese)
[36]Zhang Y,Hu XH,Shi Y,et al.Beneficial role of exogenous spermidine on nitrogen metabolism in tomato seedlings exposed to saline-alkaline stress.Journal of American Society for Horticultural Science,2013,138:38-49
[37]Wang C-Y(王春燕),Li J-R(李敬蕊),Xia Q-P(夏庆平),et al.Influence of exogenousγ-aminobutyric acid(GABA)on GABA metabolism and amino acid contents in roots of melon seedling under hypoxia stress.Chinese Journal of Applied Ecology(应用生态学报),2014,25(7):2011-2018(in Chinese)
[38]Hu XH,Xu ZR,Xu WN,et al.Application ofγ-aminobutyric acid demonstrates a protective role of polyamine and GABA metabolism in muskmelon seedlings under Ca(NO3)2stress.Plant Physiology and Biochemistry,2015,92:1-10
[2]Hu K-L(胡克玲),Wang J-T(汪季涛),Chen Y-G(陈友根),et al.Selection of salt tolerance in melon(Cucumis melo L.)at germination stage.Chinese Journal of Tropical Crops(热带作物学报),2013,34(2):228-231(in Chinese)
[3]Hell R,Mendel RR.Cell biology of metals and nutrients//Hell R,Mendel RR,eds.Plant Cell Monographs.Berlin:Springer-Verlag,2010:17
[4]Tian Y-Q(田永强),Wang J-G(王敬国),Gao L-H(高丽红).Research progress on vegetable soil microbial obstacles in protected cropping systems.China Vegetables(中国蔬菜),2013(20):1-9(in Chinese)
[5]Yuan LY,Du J,Yuan YH,et al.Effects of 24-epibrassinolide on ascorbate-glutathione cycle and polyamine levels in cucumber roots under Ca(NO3)2stress.Acta Physiologiae Plantarum,2013,35:253-262
[6]Zhou W-H(周万海),Shi S-L(师尚礼),Kou J-T(寇江涛).Effects of exogenous nitric oxide on the growth and nitrogen metabolism of alfalfa seedlings under salt stress.Chinese Journal of Applied Ecology(应用生态学报),2012,23(11):3003-3008(in Chinese)
[7]Surabhi GK,Reddy AM,Kumari GJ,et al.Modulations in key enzymes of nitrogen metabolism in two high yielding genotypes of mulberry(Morus alba L.)with differential sensitivity to salt stress.Environmental and Experimental Botany,2008,64:171-179
[8]Li JM,Hu LP,Zhang L,et al.Exogenous spermidine is enhancing tomato tolerance to salinity-alkalinity stress by regulating chloroplast antioxidant system and chlorophyll metabolism.BMC Plant Biology,2015,15:303
[9]Xiang L-X(向丽霞),Hu L-P(胡立盼),Hu X-H(胡晓辉),et al.Response of reactive oxygen metabolism in melon chloroplasts to short-term salinity-alkalinity stress regulated by exogenousγ-aminobutyric acid.Chinese Journal of Applied Ecology(应用生态学报),2015,26(12):3746-3752(in Chinese)
[10]Wang YS,Luo ZS,Huang XD,et al.Effect of exogenousγ-aminobutyric acid(GABA)treatment on chilling injury and antioxidant capacity in banana peel.Scientia Horticulturae,2014,168:132-137
[11]Barbosa JM,Singh NK,Cherry JH,et al.Nitrate uptake and utilization is modulated by exogenousγ-aminobutyric acid in Arabidopsis thaliana seedlings.Plant Physiology and Biochemistry,2010,49:443-450
[12]Bouche N,Lacombe B,Fromm H.GABA signaling:A conserved and ubiquitous mechanism.Trends in Cell Biology,2003,13:607-610
[13]Fas P,Niogret MF,Montes E,et al.Transcriptional profiling of genes encoding GABA-transaminases in Brassica napus reveals their regulation by water deficit.Environmental and Experimental Botany,2015,116:20-31
[14]Xia Q-P(夏庆平),Gao H-B(高洪波),Li J-R(李敬蕊).Effects ofγ-aminobutyric acid on the photosynthesis and chlorophyll fluorescence parameters of muskmelon seedlings under hypoxia stress.Chinese Journal of Applied Ecology(应用生态学报),2011,22(4):999-1006(in Chinese)
[15]Fan L-Q(范龙泉),Yang L-W(杨丽文),Gao H-B(高洪波),et al.Effects of exogenousγ-aminobutyric acid on polyamine metabolism of melon seedlings under hypoxia stress.Chinese Journal of Applied Ecology(应用生态学报),2012,23(6):1599-1606(in Chinese)
[16]Xu Z-R(徐志然),Zhang L(张丽),Zhang Z(张智),et al.Effects of GABA on tolerance of muskmelon seedlings against Ca(NO3)2stress.Journal of Northwest A&F University(Natural Science)(西北农林科技大学学报:自然科学版),2015,43(3):125-131(in Chinese)
[17]Wu G-P(魏国平),Zhu Y-L(朱月林),Liu Z-L(刘正鲁),et al.Effects of calcium nitrate stress on ascorbate-glutathione cycle metabolism in leaves of hydroponically-grown grafted eggplant seedlings.Chinese Journal of Plant Ecology(植物生态学报),2008,32(5):1023-1030(in Chinese)
[18]Li H-S(李合生).Principle and Technology of Plant Physiological and Biochemical Experiments.Beijing:Higher Education Press,2000(in Chinese)
[19]Tang Z-C(汤章城).Modern Plant Physiology Laboratory Manual.Beijing:Science Press,1999(in Chinese)
[20]Singh RD,Srivastava HS.Increase in glutamate synthase(NADH)activity in maize seedling in response to nitrate and ammonium nitrogen.Plant Physiology,1986,66:413-416
[21]Kanamori T,Konishi S,Takahashi E.Inducible formation of glutamate dehydrogenase in rice plant roots by the addition of ammonia to the media.Plant Physiology,1972,26:1-6
[22]Bradford MM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry,1976,72:248-254
[23]Zhang GJ,Bown AW.The rapid determination ofγ-aminobutyric acid.Phytochemistry,1997,44:1007-1009
[24]Tong H(童辉),Zhang Z-X(张振兴),Li B(李斌),et al.Effects of iso-osmotic Ca(NO3)2and Na Cl stress on chloroplast ultrastructure and photosynthesis in cucumber leaves.China Vegetables(中国蔬菜),2012(18):160-165(in Chinese)
[25]Yuan LY,Shu S,Sun J.Effects of 24-epibrassinolide on the photosynthetic characteristics,antioxidant system,and chloroplast ultrastructure in Cucumis sativus L.under Ca(NO3)2stress.Photosynthesis Research,2012,112:205-214
[26]Huang J(黄娟),Li X-F(李兴发),Huang S(黄山),et al.Effects of GABA soaking on cucumber seed germination and seedling growth under different temperature stress.Journal of Changjiang Vegetables(长江蔬菜),2014(12):30-35(in Chinese)
[27]Gao H-B(高洪波),Zhang T-J(章铁军),Lv G-Y(吕桂云),et al.Effects of exogenousγ-aminobutyric acid on growth and reactive oxygen species metabolism of cucumber seedlings under Na Cl stress.Acta Botanica Boreali-Occidentalia Sinica(西北植物学报),2007,27(10):2046-2051(in Chinese)
[28]Gangwar S,Singh VP.Indole acetic acid differently changes growth and nitrogen metabolism in Pisum sativum L.seedlings under chromium(Vl)phytotoxicity:Implication of oxidative stress.Scientia Horticulturae,2011,129:321-32
[29]Zhang Y(张毅),Shi Y(石玉),Hu X-H(胡晓辉),et al.Effects of exogenous spermidine on nitrogen metabolism and contents of main mineral elements in tomato seedlings under salinity-alkalinity mixed stress.Chinese Journal of Applied Ecology(应用生态学报),2013,24(5):1401-1408(in Chinese)
[30]Song S-L(宋锁玲),Li J-R(李敬蕊),Gao H-B(高洪波),et al.Effects of exogenousγ-aminobutyric acid on inorganic nitrogen metabolism and mineral elements contents of melon seedling under hypoxia stress.Acta Horticulturae Sinica(园艺学报),2012,39(4):695-704(in Chinese)
[31]Wang XP,Geng SJ,Ri YJ,et al.Physiological responses and adaptive strategies of tomato plants to salt and alkali stresses.Scientia Horticulturae,2011,130:248-255
[32]Li B(李冰),Zhang Z-G(张照贵),Wang J-J(王佳佳),et al.Cloning and functional marker of GDH1gene in wheat.Shandong Agricultural Sciences(山东农业科学),2014,46(10):6-11(in Chinese)
[33]Liang CG,Chen LP,Wang Y,et al.High temperature at grain-filling stage affects nitrogen metabolism enzyme activities in grains and grain nutritional quality in rice.Rice Science,2011,18:210-216
[34]Ferreira-Silva SL,Silva EN,Carvalho FEL,et al.Physiological alterations modulated by rootstock and scion combination in cashew under salinity.Hortscience,2010,127:39-45
[35]Kang Y-Y(康云艳),Guo S-R(郭世荣),Duan J-J(段九菊).Effects of root zone hypoxia on respiratory metabolism of cucumber seedlings roots.Chinese Journal of Applied Ecology(应用生态学报),2008,19(3):583-587(in Chinese)
[36]Zhang Y,Hu XH,Shi Y,et al.Beneficial role of exogenous spermidine on nitrogen metabolism in tomato seedlings exposed to saline-alkaline stress.Journal of American Society for Horticultural Science,2013,138:38-49
[37]Wang C-Y(王春燕),Li J-R(李敬蕊),Xia Q-P(夏庆平),et al.Influence of exogenousγ-aminobutyric acid(GABA)on GABA metabolism and amino acid contents in roots of melon seedling under hypoxia stress.Chinese Journal of Applied Ecology(应用生态学报),2014,25(7):2011-2018(in Chinese)
[38]Hu XH,Xu ZR,Xu WN,et al.Application ofγ-aminobutyric acid demonstrates a protective role of polyamine and GABA metabolism in muskmelon seedlings under Ca(NO3)2stress.Plant Physiology and Biochemistry,2015,92:1-10