桃PpSnRK1α在番茄中超表达提高植株耐盐性
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摘要
以超表达桃(Prunus persica)蔗糖非发酵蛋白激酶-1 (SnRK1)基因(Pp Sn RK1α)的番茄(Solanum lycopersicum)株系及野生型为试材,研究盐胁迫下SnRK1对植株生长的影响。结果表明:盐胁迫下,番茄叶片和根系中SnRK1活性呈现先上升后下降的趋势,但超表达PpSnRK1α番茄叶片和根系中SnRK1活性始终显著高于野生型植株;正常条件下,超表达PpSnRK1α番茄相比野生型根系活力提高了10.16%,盐胁迫下提高了18.92%,差异显著;叶片伊文思蓝(Evans blue)染色结果发现,超表达PpSnRK1α番茄叶片受伤害程度明显轻于野生型;盐胁迫处理3、6、9和12 d后,超表达PpSnRK1α番茄叶片过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均显著高于野生型植株,而超氧阴离子(O_2ˉ)和丙二醛含量显著低于野生型植株;盐胁迫处理4、8和12 d后,超表达PpSnRK1α番茄叶片叶绿素含量、净光合速率(P_n)及最大光化学效率(F_v/F_m)的降低幅度显著低于野生型番茄。这些结果表明,盐胁迫下,超表达PpSnRK1α番茄植株比野生型植株SnRK1活性显著提高,并通过提高植株的抗氧化能力和活性氧清除能力,缓解盐胁迫对叶片叶绿素的降解和对光系统II (PSII)的破坏程度,进而提高了植株的耐盐性。
Pp Sn RK1α-overexpressing tomato(Solanum lycopersicum) and wild type(WT) tomato were used to study the effect of sucrose non-fermenting-1-related protein kinase-1(SnRK1) on the growth of tomato plants under salt stress. The results show that, under salt stress, the activities of SnRK1 in tomato leaves and roots showed a trend of first rising and then falling, but SnRK1 activities in the leaves and roots of Pp Sn RK1α-overexpressing lines were significantly higher than those of WT. Under normal conditions, Pp Sn RK1α-overexpressing lines increased the root activity by 10.16% compared to WT. Under salt stress, it's 18.92% higher than WT, and the difference was significant. The results show that the damage degree of Pp Sn RK1α-overexpressing line leaves were significantly lower than those of WT by Evans blue dyeing. Under 3, 6, 9 and 12 days of salt stress, activities of catalase, superoxide dismutase and peroxidase in Pp Sn RK1α-overexpressing line leaves were significantly higher than those of WT, and O_2ˉ production and MDA content in PpSnRK1α-overexpressing line leaves significantly lower than those of WT. The reductions of chlorophyll content, net photosynthetic rate(Pn) and maximal photochemical efficiency of photosystem II(F_v/F_m) in Pp Sn RK1α-overexpressing lines were significantly less than those of WT. The results show that, under salt stress, SnRK1 activities in PpSnRK1α-overexpressing line leaves increased significantly than those of WT. In consequence, the abilities of oxidation resistance and reactive oxygen species removal in Pp Sn RK1α-overexpressing lines were improved, the degradation of chlorophyll and the degree of damage to the photosystem II were alleviated, and salt resistance of the plants was improved.
Pp Sn RK1α-overexpressing tomato(Solanum lycopersicum) and wild type(WT) tomato were used to study the effect of sucrose non-fermenting-1-related protein kinase-1(SnRK1) on the growth of tomato plants under salt stress. The results show that, under salt stress, the activities of SnRK1 in tomato leaves and roots showed a trend of first rising and then falling, but SnRK1 activities in the leaves and roots of Pp Sn RK1α-overexpressing lines were significantly higher than those of WT. Under normal conditions, Pp Sn RK1α-overexpressing lines increased the root activity by 10.16% compared to WT. Under salt stress, it's 18.92% higher than WT, and the difference was significant. The results show that the damage degree of Pp Sn RK1α-overexpressing line leaves were significantly lower than those of WT by Evans blue dyeing. Under 3, 6, 9 and 12 days of salt stress, activities of catalase, superoxide dismutase and peroxidase in Pp Sn RK1α-overexpressing line leaves were significantly higher than those of WT, and O_2ˉ production and MDA content in PpSnRK1α-overexpressing line leaves significantly lower than those of WT. The reductions of chlorophyll content, net photosynthetic rate(Pn) and maximal photochemical efficiency of photosystem II(F_v/F_m) in Pp Sn RK1α-overexpressing lines were significantly less than those of WT. The results show that, under salt stress, SnRK1 activities in PpSnRK1α-overexpressing line leaves increased significantly than those of WT. In consequence, the abilities of oxidation resistance and reactive oxygen species removal in Pp Sn RK1α-overexpressing lines were improved, the degradation of chlorophyll and the degree of damage to the photosystem II were alleviated, and salt resistance of the plants was improved.
引文
Ahmad P,Sharma S(2010).Physio-biochemical attributes in two cultivars of mulberry(Morus alba L.)under NaHCO3stress.Int J Plant Prod,4(2):79-86
Baena-González E,Rolland F,Thevelein JM,et al(2007).Acentral integrator of transcription networks in plant stress and energy signalling.Nature,448:938-942
Cai Z,Cao K,Feng Y,et al(2003).Effect of low nocturnal temperature stress on fluorescence characteristics and active oxygen metabolism in leaves of Garcinia hanburyi seedlings grown under two levels of irradiance.Chin JAppl Ecol,14(3):326-330(in Chinese with English abstract)[蔡志全,曹坤芳,冯玉龙等(2003).夜间低温胁迫对两种生长光强下藤黄幼苗叶片荧光特征和活性氧代谢的影响.应用生态学报,14(3):326-330]
Chen YZ,Patterson BD(1988).The effect of chilling temperature on the level of superoxide dismutase,catalase and hydrogen peroxide in some plant leaves.Acta Phytophysiol Sin,14:323-328(in Chinese with English abstract)[陈贻竹,帕特森(1988).低温对植物叶片中超氧物歧化酶、过氧化物酶和过氧化氢水平的影响.植物生理学报,14:323-328]
Crozet P,Jammes F,Valot B,et al(2010).Cross-phosphorylation between Arabidopsis thaliana Sucrose Nonfermenting 1-related Protein Kinase 1(AtSnRK1)and its activating kinase(AtSnAK)determines their catalytic activities.J Biol Chem,285:12071-12077
Debast S,Nunes-Nesi A,Hajirezaei MR,et al(2011).Altering trehalose-6-phosphate content in transgenic potato tubers growth and alters responsiveness to hormones during sprouting.Plant Physiol,156:1754-1771
Geigenberger P,Stitt M,Fernie AR(2004).Metabolic control analysis and regulation of the conversion of sucrose to starch in growing potato tubers.Plant Cell Environ,27(6):655-673
Halford NG,Hardie DG(1998).SNF1-related protein kinases:global regulators of carbon metabolism in plants?Plant Mol Biol,37:735-748
Hulsmans S,Rodriguez M,De Coninck B,et al(2016).The SnRK1 energy sensor in plant biotic interactions.Trends Plant Sci,21(8):648-661
Kar M,Mishra D(1976).Catalase,peroxidase,and polyphenoloxidase activities during rice leaf senescence.Plant Physiol,57(2):315-319
Li PM,Gao HY,Strasser RJ(2005).Application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study.J Plant Physiol Mol Biol,31(6):559-566(in Chinese with English abstract)[李鹏民,高辉远,Strasser RJ(2005).快速叶绿素荧光诱导动力学分析在光合作用研究中的应用.植物生理与分子生物学学报,31(6):559-566]
Liu JX,Wang JC,Wang RJ,et al(2015).Effects of salt and alkali stress on photosynthesis in Avena nuda seedlings.Agr Res Arid Areas,33(6):155-160(in Chinese with English abstract)[刘建新,王金成,王瑞娟等(2015).盐、碱胁迫对燕麦幼苗光合作用的影响.干旱地区农业研究,33(6):155-160]
Liu JX,Wang X,Wang RJ,et al(2011).Effects of alkaline stress on the metabolism of reactive oxygen species and osmotica accumulation in ryegrass seedling roots.Bull Bot Res,31(6):674-679(in Chinese with English abstract)[刘建新,王鑫,王瑞娟等(2011).碱胁迫对黑麦草幼苗根系活性氧代谢和渗透溶质积累的影响.植物研究,31(6):674-679]
Liu N,Lin ZF(2011).Use of Evans blue for testing cell viability of intact leaves of plant.Plant Physiol J,47(6):570-574(in Chinese with English abstract)[刘楠,林植芳(2011).用伊文思蓝染色法检测植物整体叶片的细胞活性.植物生理学报,47(6):570-574]
Liu XJ,Liu X,An XH,et al(2017).An apple protein kinase MdSnRK1.1 interacts with MdCAIP1 to regulate ABAsensitivity.Plant Cell Physiol,58:1631-1641
Luo J,Zhao Y,Peng Y,et al(2017).Effects of overexpressing Pp Sn RK1αon the growth of tomato plants under nutrient stress.Acta Hortic Sin,44(4):644-652(in Chinese with English abstract)[罗静静,张亚飞,彭妍等(2017).桃PpSnRK1α在番茄中过表达对营养胁迫下植株生长的影响.园艺学报,44(4):644-652]
Luo JJ,Zhnag YF,Zhao YF,et al(2018).Effects of salicylic acid on the SnRK1 activity and growth of strawberry plant.Plant Physiol J,54(1):113-120(in Chinese with English abstract)[罗静静,张亚飞,赵永飞等(2018).水杨酸对草莓SnRK1活性及植株生长的影响.植物生理学报,54(1):113-120]
Nukarinen E,N?gele T,Pedrotti L,et al(2016).Quantitative phosphoproteomics reveals the role of the AMPK plant ortholog SnRK1 as a metabolic master regulator under energy deprivation.Sci Rep,6:31697
Omran RG(1980).Peroxide levels and the activities of catalase,peroxidase,and indoleacetic acid oxidase during and after chilling cucumber seedlings.Plant Physiol,65(2):407-408
Radchuk R,Radchk V,Weschke W,et al(2006).Repressing the expression of the SUCR OSE NONFERMENT-I N G-1-R E L AT E D P R OT E I N K I N A S E g e n e i n p e a embryo causes pleiotropic defects of maturation similar to an abscisic acid-insensitive phenotype.Plant Physiol,140:263-278
Rodrigues A,Adamo M,Crozet P,et al(2013).ABI1 and PP2CA phosphatases are negative regulators of Snf1-Related Protein Kinase1 signaling in Arabidopsis.Plant Cell,25:3871-3884
Smeekens S,Ma J,Hanson J,et al(2010).Sugar signals and molecular networks controlling plant growth.Curr Opin Plant Biol,13:274-279
Wang GF,Peng FT,Zhang YF,et al(2014).Effects of overexpressing Pingyi Tiancha MhSnRK1 on carbohydrate metabolism in tomato.Acta Hortic Sin,41(11):2188-2195(in Chinese with English abstract)[王贵芳,彭福田,张亚飞等(2014).平邑甜茶MhSnRK1在番茄中超表达对植株碳代谢的影响.园艺学报,41(11):2188-2195]
Wang X,Peng F,Li M,et al(2012).Expression of a heterologous SnRK1 in tomato increases carbon assimilation,nitrogen uptake and modifies fruit development.J Plant Physiol,169:1173-1182
Zhang Y,Primavesi LF,Jhurreea D,et al(2009).Inhibition of SNF1-related protein kinase1 activity and regulation of metabolic pathways by trehalose-6-phosphate.Plant Physiol,149:1860-1871
Zhao S,Shi G,Dong X(2002).Techniques of Plant Physiological Experiment.Beijing:Chinese Agricultural Science and Technology Press(in Chinese)[赵世杰,史国安,董新纯(2002).植物生理学实验指导.北京:中国农业科学技术出版社]
Zhao YF,Chen XL,Peng FT,et al(2017).Over expression of a peach Pp Sn RK1βγ1 gene improving oxidative stress tolerance in Arabidopsis thaliana.Plant Physiol J,53(7):1251-1258(in Chinese with English abstract)[赵永飞,陈晓璐,彭福田等(2017).桃PpSnRK1βγ1在拟南芥中超表达提高植株氧化胁迫耐性.植物生理学报,53(7):1251-1258]
Zimmermann P,Heinlein C,Orendi G,et al(2006).Senescence-specific regulation of catalases in Arabidopsis thaliana(L.)Heynh.Plant Cell Environ,29(6):1049-1060
Baena-González E,Rolland F,Thevelein JM,et al(2007).Acentral integrator of transcription networks in plant stress and energy signalling.Nature,448:938-942
Cai Z,Cao K,Feng Y,et al(2003).Effect of low nocturnal temperature stress on fluorescence characteristics and active oxygen metabolism in leaves of Garcinia hanburyi seedlings grown under two levels of irradiance.Chin JAppl Ecol,14(3):326-330(in Chinese with English abstract)[蔡志全,曹坤芳,冯玉龙等(2003).夜间低温胁迫对两种生长光强下藤黄幼苗叶片荧光特征和活性氧代谢的影响.应用生态学报,14(3):326-330]
Chen YZ,Patterson BD(1988).The effect of chilling temperature on the level of superoxide dismutase,catalase and hydrogen peroxide in some plant leaves.Acta Phytophysiol Sin,14:323-328(in Chinese with English abstract)[陈贻竹,帕特森(1988).低温对植物叶片中超氧物歧化酶、过氧化物酶和过氧化氢水平的影响.植物生理学报,14:323-328]
Crozet P,Jammes F,Valot B,et al(2010).Cross-phosphorylation between Arabidopsis thaliana Sucrose Nonfermenting 1-related Protein Kinase 1(AtSnRK1)and its activating kinase(AtSnAK)determines their catalytic activities.J Biol Chem,285:12071-12077
Debast S,Nunes-Nesi A,Hajirezaei MR,et al(2011).Altering trehalose-6-phosphate content in transgenic potato tubers growth and alters responsiveness to hormones during sprouting.Plant Physiol,156:1754-1771
Geigenberger P,Stitt M,Fernie AR(2004).Metabolic control analysis and regulation of the conversion of sucrose to starch in growing potato tubers.Plant Cell Environ,27(6):655-673
Halford NG,Hardie DG(1998).SNF1-related protein kinases:global regulators of carbon metabolism in plants?Plant Mol Biol,37:735-748
Hulsmans S,Rodriguez M,De Coninck B,et al(2016).The SnRK1 energy sensor in plant biotic interactions.Trends Plant Sci,21(8):648-661
Kar M,Mishra D(1976).Catalase,peroxidase,and polyphenoloxidase activities during rice leaf senescence.Plant Physiol,57(2):315-319
Li PM,Gao HY,Strasser RJ(2005).Application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study.J Plant Physiol Mol Biol,31(6):559-566(in Chinese with English abstract)[李鹏民,高辉远,Strasser RJ(2005).快速叶绿素荧光诱导动力学分析在光合作用研究中的应用.植物生理与分子生物学学报,31(6):559-566]
Liu JX,Wang JC,Wang RJ,et al(2015).Effects of salt and alkali stress on photosynthesis in Avena nuda seedlings.Agr Res Arid Areas,33(6):155-160(in Chinese with English abstract)[刘建新,王金成,王瑞娟等(2015).盐、碱胁迫对燕麦幼苗光合作用的影响.干旱地区农业研究,33(6):155-160]
Liu JX,Wang X,Wang RJ,et al(2011).Effects of alkaline stress on the metabolism of reactive oxygen species and osmotica accumulation in ryegrass seedling roots.Bull Bot Res,31(6):674-679(in Chinese with English abstract)[刘建新,王鑫,王瑞娟等(2011).碱胁迫对黑麦草幼苗根系活性氧代谢和渗透溶质积累的影响.植物研究,31(6):674-679]
Liu N,Lin ZF(2011).Use of Evans blue for testing cell viability of intact leaves of plant.Plant Physiol J,47(6):570-574(in Chinese with English abstract)[刘楠,林植芳(2011).用伊文思蓝染色法检测植物整体叶片的细胞活性.植物生理学报,47(6):570-574]
Liu XJ,Liu X,An XH,et al(2017).An apple protein kinase MdSnRK1.1 interacts with MdCAIP1 to regulate ABAsensitivity.Plant Cell Physiol,58:1631-1641
Luo J,Zhao Y,Peng Y,et al(2017).Effects of overexpressing Pp Sn RK1αon the growth of tomato plants under nutrient stress.Acta Hortic Sin,44(4):644-652(in Chinese with English abstract)[罗静静,张亚飞,彭妍等(2017).桃PpSnRK1α在番茄中过表达对营养胁迫下植株生长的影响.园艺学报,44(4):644-652]
Luo JJ,Zhnag YF,Zhao YF,et al(2018).Effects of salicylic acid on the SnRK1 activity and growth of strawberry plant.Plant Physiol J,54(1):113-120(in Chinese with English abstract)[罗静静,张亚飞,赵永飞等(2018).水杨酸对草莓SnRK1活性及植株生长的影响.植物生理学报,54(1):113-120]
Nukarinen E,N?gele T,Pedrotti L,et al(2016).Quantitative phosphoproteomics reveals the role of the AMPK plant ortholog SnRK1 as a metabolic master regulator under energy deprivation.Sci Rep,6:31697
Omran RG(1980).Peroxide levels and the activities of catalase,peroxidase,and indoleacetic acid oxidase during and after chilling cucumber seedlings.Plant Physiol,65(2):407-408
Radchuk R,Radchk V,Weschke W,et al(2006).Repressing the expression of the SUCR OSE NONFERMENT-I N G-1-R E L AT E D P R OT E I N K I N A S E g e n e i n p e a embryo causes pleiotropic defects of maturation similar to an abscisic acid-insensitive phenotype.Plant Physiol,140:263-278
Rodrigues A,Adamo M,Crozet P,et al(2013).ABI1 and PP2CA phosphatases are negative regulators of Snf1-Related Protein Kinase1 signaling in Arabidopsis.Plant Cell,25:3871-3884
Smeekens S,Ma J,Hanson J,et al(2010).Sugar signals and molecular networks controlling plant growth.Curr Opin Plant Biol,13:274-279
Wang GF,Peng FT,Zhang YF,et al(2014).Effects of overexpressing Pingyi Tiancha MhSnRK1 on carbohydrate metabolism in tomato.Acta Hortic Sin,41(11):2188-2195(in Chinese with English abstract)[王贵芳,彭福田,张亚飞等(2014).平邑甜茶MhSnRK1在番茄中超表达对植株碳代谢的影响.园艺学报,41(11):2188-2195]
Wang X,Peng F,Li M,et al(2012).Expression of a heterologous SnRK1 in tomato increases carbon assimilation,nitrogen uptake and modifies fruit development.J Plant Physiol,169:1173-1182
Zhang Y,Primavesi LF,Jhurreea D,et al(2009).Inhibition of SNF1-related protein kinase1 activity and regulation of metabolic pathways by trehalose-6-phosphate.Plant Physiol,149:1860-1871
Zhao S,Shi G,Dong X(2002).Techniques of Plant Physiological Experiment.Beijing:Chinese Agricultural Science and Technology Press(in Chinese)[赵世杰,史国安,董新纯(2002).植物生理学实验指导.北京:中国农业科学技术出版社]
Zhao YF,Chen XL,Peng FT,et al(2017).Over expression of a peach Pp Sn RK1βγ1 gene improving oxidative stress tolerance in Arabidopsis thaliana.Plant Physiol J,53(7):1251-1258(in Chinese with English abstract)[赵永飞,陈晓璐,彭福田等(2017).桃PpSnRK1βγ1在拟南芥中超表达提高植株氧化胁迫耐性.植物生理学报,53(7):1251-1258]
Zimmermann P,Heinlein C,Orendi G,et al(2006).Senescence-specific regulation of catalases in Arabidopsis thaliana(L.)Heynh.Plant Cell Environ,29(6):1049-1060