低温下冬小麦品种间甜菜碱及其代谢途径比较
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摘要
为研究甜菜碱以及其关键合成酶甜菜碱醛脱氢酶在低温胁迫时对小麦的保护作用,笔者将抗寒性不同的2个冬小麦品种‘东农冬麦一号’(DM1)和‘济麦22’(J22)栽培于自然条件下,通过测定苗期相对电导率、丙二醛(MDA)含量、甜菜碱醛脱氢酶活性、甜菜碱醛脱氢酶基因表达量以及甜菜碱含量。结果发现,抗寒品种DM1表现出甜菜碱醛脱氢酶酶活和甜菜碱含量大幅增加,而封冻期MDA含量相比J22却有所降低。这说明甜菜碱醛脱氢酶基因的表达促进了甜菜碱的积累,同时增加了细胞膜的稳定性。本研究结果不仅确定了甜菜碱在DM1抗寒性中的作用,同时为今后提高冬小麦抗寒性提供新的研究方向。
The paper aims to study glycine betaine(GB) and the protection effect of betaine aldehyde dehydrogenase(BADH), which is the key synthetase, on wheat. The author planted two winter wheat varietieswith different cold resistance,‘Dongnongdongmai1'(DM1) and‘Jimai22'(J22) under natural condition, andanalyzed the relative electrical conductivity, malondialdehyde(MDA) content, BADH activity, the expressionof BADH, and the content of GB at the seedling stage. The results showed that: DM1 presented a great increasein BADH activity and GB content, but a decrease in MDA content under freezing condition compared with J22,indicating that the expression of BADH promoted the accumulation of glycine betaine and increased thestability of cell membrane. The results of this study not only confirm the role of glycine betaine in coldresistance of DM1, but also provide a new research direction for improving the cold resistance of winter wheat in the future.
The paper aims to study glycine betaine(GB) and the protection effect of betaine aldehyde dehydrogenase(BADH), which is the key synthetase, on wheat. The author planted two winter wheat varietieswith different cold resistance,‘Dongnongdongmai1'(DM1) and‘Jimai22'(J22) under natural condition, andanalyzed the relative electrical conductivity, malondialdehyde(MDA) content, BADH activity, the expressionof BADH, and the content of GB at the seedling stage. The results showed that: DM1 presented a great increasein BADH activity and GB content, but a decrease in MDA content under freezing condition compared with J22,indicating that the expression of BADH promoted the accumulation of glycine betaine and increased thestability of cell membrane. The results of this study not only confirm the role of glycine betaine in coldresistance of DM1, but also provide a new research direction for improving the cold resistance of winter wheat in the future.
引文
[1]刘雨译.施加甜菜碱对促进体外培养的茶树嫩枝增殖和溶液摄取率的研究[J].南方农药,2014(3):45-50.
[2] Yancey P H, Clark M E, Hand S C, et al. Living with Water Stress:Evolution of Osmolyte Systems[J].Science,1982,217(4566):1214.
[3]梁超.过量积累甜菜碱改善小麦耐盐性的生理机制研究[D].泰安:山东农业大学,2007:14-15.
[4] Yang C, Zhou Y, Fan J, et al. SpBADH of the halophyte Sesuvium portulacastrum strongly confers drought tolerance through ROS scavenging in transgenic Arabidopsis[J]. Plant Physiology&Biochemistry,2015,96:377-387.
[5] Fan W, Min Z, Zhang H, et al. Improved Tolerance to Various Abiotic Stresses in Transgenic Sweet Potato(Ipomoea batatas)Expressing Spinach Betaine Aldehyde Dehydrogenase[J]. Plos One,2012,7(5):e37344.
[6]张兰峰,刘雅帧,吴泽华.甜菜碱的生产方法及应用[J].生物技术世界,2012(5):32-33.
[7]郭岩,张莉,肖岗,等.甜菜碱醛脱氢酶基因在水稻中的表达及转基因植株的耐盐性研究[J].中国科学:生命科学,1997,27(2):151-155.
[8]熊方杰.共表达盐生隐杆藻甘氨酸甲基化酶基因ApGSMT和ApDMT提高水稻抗逆性研究[D].重庆:重庆大学,2013:11-12.
[9]纪璎.反应性羧酸甜菜碱两性离子共聚物构建皮细胞选择性功能界面研究[D].杭州:浙江大学,2012:13-14.
[10] Rathinasabapathi B, Mccue K F, Gage D A, et al. Metabolic engineering of glycine betaine synthesis:plant betaine aldehyde dehydrogenases lacking typical transit peptides are targeted to tobacco chloroplasts where they confer betaine aldehyde resistance[J]. Planta,1994,193(2):155-162.
[11]梁峥,马德钦,汤岚,等.菠菜甜菜碱醛脱氢酶基因在烟草中的表达[J].生物工程学报,1997,13(3):236-240.
[12]刘凤华,郭岩,谷冬梅,等.转甜菜碱醛脱氢酶基因植物的耐盐性研究[J].遗传学报,1997(1):54-58.
[13]李枝梅,窦海鸥,卫丹丹,等.转codA基因提高番茄植株的耐热性[J].作物学报,2013,39(11):2046-2054.
[14]蒋玉蓉,袁俊杰,陈国林,等.转甜菜碱醛脱氢酶(BADH)基因棉花的获得及其耐盐性鉴定[J].分子植物育种,2015,13(1):125-131.
[15] Holmstr?m K O, Welin B, Mandal A, et al. Production of the Escherichia coli betaine-aldehyde dehydrogenase, an enzyme required for the synthesis of the osmoprotectant glycine betaine, in transgenic plants[J].Plant Journal,1994,6(5):749.
[16] Sakamoto A. Enhancement of the tolerance of Arabidopsis to high temperatures by genetic engineering of the synthesis of glycinebetaine[J].Plant Journal,1998,16(2):155-161.
[17] Sakamoto A, Alia, Murata N. Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold[J].Plant Molecular Biology,1998,38(6):1011-1019.
[18] Sakamoto A, Murata N. Genetic engineering of glycinebetaine synthesis in plants:current status and implications for enhancement of stress tolerance[J]. Journal of Experimental Botany,2000,51(342):81.
[19]陈传芳,李义文,陈豫,等.通过农杆菌介导法获得耐盐转甜菜碱醛脱氢酶基因白三叶草[J].遗传学报,2004,31(1):97-101.
[20]郝再彬.植物生理实验[M].哈尔滨:哈尔滨工业大学出版社,2006:101-104.
[21]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:260-261.
[22]罗笛,牛向丽,余进德,等.水稻、菠菜甜菜碱醛脱氢酶(BADH)重组蛋白表达及酶活性分析[J].农业生物技术学报,2012,20(12):1390-1397.
[23]齐永秀,郝立勇,翟宏艳,等.非水滴定法测定盐酸甜菜碱的含量[J].泰山医学院学报,2002,23(4):355-356.
[24]黄丽华.甜菜碱对玉米幼苗抗寒性的影响[J].湖北农业科学,2006(2):168-170.
[25]陈文超,杨博智,周书栋,等.3种诱导剂对辣椒幼苗抗寒性的影响[J].湖南农业大学学报:自然科学版,2011,37(4):396-399.
[2] Yancey P H, Clark M E, Hand S C, et al. Living with Water Stress:Evolution of Osmolyte Systems[J].Science,1982,217(4566):1214.
[3]梁超.过量积累甜菜碱改善小麦耐盐性的生理机制研究[D].泰安:山东农业大学,2007:14-15.
[4] Yang C, Zhou Y, Fan J, et al. SpBADH of the halophyte Sesuvium portulacastrum strongly confers drought tolerance through ROS scavenging in transgenic Arabidopsis[J]. Plant Physiology&Biochemistry,2015,96:377-387.
[5] Fan W, Min Z, Zhang H, et al. Improved Tolerance to Various Abiotic Stresses in Transgenic Sweet Potato(Ipomoea batatas)Expressing Spinach Betaine Aldehyde Dehydrogenase[J]. Plos One,2012,7(5):e37344.
[6]张兰峰,刘雅帧,吴泽华.甜菜碱的生产方法及应用[J].生物技术世界,2012(5):32-33.
[7]郭岩,张莉,肖岗,等.甜菜碱醛脱氢酶基因在水稻中的表达及转基因植株的耐盐性研究[J].中国科学:生命科学,1997,27(2):151-155.
[8]熊方杰.共表达盐生隐杆藻甘氨酸甲基化酶基因ApGSMT和ApDMT提高水稻抗逆性研究[D].重庆:重庆大学,2013:11-12.
[9]纪璎.反应性羧酸甜菜碱两性离子共聚物构建皮细胞选择性功能界面研究[D].杭州:浙江大学,2012:13-14.
[10] Rathinasabapathi B, Mccue K F, Gage D A, et al. Metabolic engineering of glycine betaine synthesis:plant betaine aldehyde dehydrogenases lacking typical transit peptides are targeted to tobacco chloroplasts where they confer betaine aldehyde resistance[J]. Planta,1994,193(2):155-162.
[11]梁峥,马德钦,汤岚,等.菠菜甜菜碱醛脱氢酶基因在烟草中的表达[J].生物工程学报,1997,13(3):236-240.
[12]刘凤华,郭岩,谷冬梅,等.转甜菜碱醛脱氢酶基因植物的耐盐性研究[J].遗传学报,1997(1):54-58.
[13]李枝梅,窦海鸥,卫丹丹,等.转codA基因提高番茄植株的耐热性[J].作物学报,2013,39(11):2046-2054.
[14]蒋玉蓉,袁俊杰,陈国林,等.转甜菜碱醛脱氢酶(BADH)基因棉花的获得及其耐盐性鉴定[J].分子植物育种,2015,13(1):125-131.
[15] Holmstr?m K O, Welin B, Mandal A, et al. Production of the Escherichia coli betaine-aldehyde dehydrogenase, an enzyme required for the synthesis of the osmoprotectant glycine betaine, in transgenic plants[J].Plant Journal,1994,6(5):749.
[16] Sakamoto A. Enhancement of the tolerance of Arabidopsis to high temperatures by genetic engineering of the synthesis of glycinebetaine[J].Plant Journal,1998,16(2):155-161.
[17] Sakamoto A, Alia, Murata N. Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold[J].Plant Molecular Biology,1998,38(6):1011-1019.
[18] Sakamoto A, Murata N. Genetic engineering of glycinebetaine synthesis in plants:current status and implications for enhancement of stress tolerance[J]. Journal of Experimental Botany,2000,51(342):81.
[19]陈传芳,李义文,陈豫,等.通过农杆菌介导法获得耐盐转甜菜碱醛脱氢酶基因白三叶草[J].遗传学报,2004,31(1):97-101.
[20]郝再彬.植物生理实验[M].哈尔滨:哈尔滨工业大学出版社,2006:101-104.
[21]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:260-261.
[22]罗笛,牛向丽,余进德,等.水稻、菠菜甜菜碱醛脱氢酶(BADH)重组蛋白表达及酶活性分析[J].农业生物技术学报,2012,20(12):1390-1397.
[23]齐永秀,郝立勇,翟宏艳,等.非水滴定法测定盐酸甜菜碱的含量[J].泰山医学院学报,2002,23(4):355-356.
[24]黄丽华.甜菜碱对玉米幼苗抗寒性的影响[J].湖北农业科学,2006(2):168-170.
[25]陈文超,杨博智,周书栋,等.3种诱导剂对辣椒幼苗抗寒性的影响[J].湖南农业大学学报:自然科学版,2011,37(4):396-399.