水氮耦合对苗期葡萄叶片氮素代谢酶活性的影响
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摘要
为研究水氮耦合对苗期葡萄叶片氮代谢影响及最佳施氮量的制定,以一年生葡萄品种红提为研究试材,利用人工控制环境的方法,在温室内采用水、氮两因素各4水平的全面设计进行实验,水分处理分别为正常灌溉W1(田间最大持水量的70%～80%)、轻度胁迫W2(60%～70%)、中度胁迫W3(50%～60%)和重度胁迫W4(30%～40%)。4个氮素施用水平分别为1.5倍推荐施肥N1(施纯氮25.5g·m~(-2))、正常推荐施肥N2(17g·m~(-2))、0.5倍推荐施肥N3(8.5g·m~(-2))、不施用氮肥N4(不施氮)。处理时间为10、20、30、40d。结果表明,在水分条件适宜时,葡萄叶片硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)活性、可溶性蛋白、游离氨基酸含量随施氮量增加而提高;在轻度干旱胁迫时,增施氮肥可缓解干旱胁迫;在重度干旱胁迫时,高氮处理使设施葡萄叶片中氮代谢酶活性、游离氨基酸和可溶性蛋白含量降低。葡萄叶片内氮含量始终随处理时间增加而降低,在轻度水分胁迫下氮的转运率较高,而水分胁迫严重时,高氮处理与无氮处理时氮转运率均偏低。最终得出:在水分条件适宜(W1)和轻度水分胁迫(W2)下,N1处理葡萄叶片的氮代谢能力最高;在中度水分胁迫(W3)和重度水分胁迫(W4)下,N3、N4处理氮代谢能力最高。研究结果可为实际生产中设施葡萄的干旱灾害防控提供理论依据,既能有效缓解水分胁迫带来的危害,又避免生产中肥料的浪费。
In order to study the effect of water and nitrogen coupling on nitrogen metabolism of grape leaves at seedling stage and the formulation of optimal nitrogen application, the annual "Hongti" grape was used as the research material. A comprehensive design of four levels of water and nitrogen in the greenhouse was used to carry out the experiment under the artificial environment control method. The water treatments were as follows: normal irrigation W1(70%-80% of maximum water capacity in the field), mild stress W2(60%-70%), moderate stress W3(50%-60%) and severe stress(30%-40%).The levels of nitrogen application were as follows:1.5 times recommended fertilization N1(pure nitrogen 25.5 g·m~(-2)), normal recommended fertilization N2(17 g·m~(-2)), 0.5 time recommended fertilization N3(8.5 g·m~(-2)), no nitrogen fertilizer N4(no nitrogen application).The treatment time was10, 20, 30, 40 days. The results showed that the content of nitrate reductase(NR), glutamine synthetase(GS),glutamine-oxoglutarate aminotransferase(GOGAT), soluble protein and free amino acid increased with the increase of nitrogen application when moisture conditions were appropriate. When drought stress was mild,increased application of nitrogen fertilizer can alleviate drought stress.When drought stress was severe, high nitrogen treatment reduced the activities of nitrogen metabolism enzymes, the content of free amino acids and soluble proteins in the leaves of the grape. The nitrogen content in the leaves of grape always decreased with the increase of treatment time. Under mild water stress, the nitrogen transport rate was higher. When the water stress was severe, the nitrogen transport rate of treatment under high nitrogen and no nitrogen were lower. Finally, under the suitable water conditions(W1) and mild water stress(W2), the nitrogen metabolism ability of grape leaves under N1(purified nitrogen 37.5 g·m~(-2)) was the highest. Under moderate water stress(W3) and severe water stress(W4), N3(pure nitrogen 12.5 g·m~(-2)) and N4(no nitrogen application) had the highest nitrogen metabolism capacity. The research results provide a theoretical basis for the prevention and control of drought disasters in the actual production of grapes, which can effectively alleviate the harm caused by water stress, and avoid the waste of fertilizer in production, so as to achieve less fertilizer efficiency.
In order to study the effect of water and nitrogen coupling on nitrogen metabolism of grape leaves at seedling stage and the formulation of optimal nitrogen application, the annual "Hongti" grape was used as the research material. A comprehensive design of four levels of water and nitrogen in the greenhouse was used to carry out the experiment under the artificial environment control method. The water treatments were as follows: normal irrigation W1(70%-80% of maximum water capacity in the field), mild stress W2(60%-70%), moderate stress W3(50%-60%) and severe stress(30%-40%).The levels of nitrogen application were as follows:1.5 times recommended fertilization N1(pure nitrogen 25.5 g·m~(-2)), normal recommended fertilization N2(17 g·m~(-2)), 0.5 time recommended fertilization N3(8.5 g·m~(-2)), no nitrogen fertilizer N4(no nitrogen application).The treatment time was10, 20, 30, 40 days. The results showed that the content of nitrate reductase(NR), glutamine synthetase(GS),glutamine-oxoglutarate aminotransferase(GOGAT), soluble protein and free amino acid increased with the increase of nitrogen application when moisture conditions were appropriate. When drought stress was mild,increased application of nitrogen fertilizer can alleviate drought stress.When drought stress was severe, high nitrogen treatment reduced the activities of nitrogen metabolism enzymes, the content of free amino acids and soluble proteins in the leaves of the grape. The nitrogen content in the leaves of grape always decreased with the increase of treatment time. Under mild water stress, the nitrogen transport rate was higher. When the water stress was severe, the nitrogen transport rate of treatment under high nitrogen and no nitrogen were lower. Finally, under the suitable water conditions(W1) and mild water stress(W2), the nitrogen metabolism ability of grape leaves under N1(purified nitrogen 37.5 g·m~(-2)) was the highest. Under moderate water stress(W3) and severe water stress(W4), N3(pure nitrogen 12.5 g·m~(-2)) and N4(no nitrogen application) had the highest nitrogen metabolism capacity. The research results provide a theoretical basis for the prevention and control of drought disasters in the actual production of grapes, which can effectively alleviate the harm caused by water stress, and avoid the waste of fertilizer in production, so as to achieve less fertilizer efficiency.
引文
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[8]Vega M,Marinod,Sanchez Z,et al.CO2 enrichment modulates ammonium nutrition in tomato adjusting carbon and nitrogen metabolism to stomatal conductance[J].Plant Sci,2015,41:32-44.
[9]丁邦琴,李铁军.谷氨酰胺高产菌株在不同NH4+浓度下的基因差异表达[J].基因组学与应用生物学,2013,32(1):41-45.Ding B Q,Li T J.Differential gene expression of high glutamine producing strain of arthrobact globiformis in the different NH4+concentration[J].Genomics and Applied Biology,2013,32(1):41-45.(in Chinese)
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[11]张华珍,徐恒玉.植物氮素同化过程中相关酶的研究进展[J].北方园艺,2011,(20):180-183.Zhang H Z,Xu H Y.Advances in the study of enzymes related to nitrogen assimilation in plants[J].Northern Horticulture,2011,(20):180-183.(in Chinese)
[12]孙永健,孙园园,蒋明金,等.施肥水平对不同氮效率水稻氮素利用特征及产量的影响[J].中国农业科学,2016,49(24):4745-4756.Sun Y J,Sun Y Y,Jiang M J,et al.Effects of fertilizer levels on nitrogen utilization characteristics and yield in rice cultivars with different nitrogen use efficiencies[J].Scientia Agricultura Sinica,2016,49(24):4745-4756.(in Chinese)
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[15]张智猛,万书波,宁堂原,等.氮素水平对花生氮素代谢及相关酶活性的影响[J].植物生态学报,2008,(6):1407-1416.Zhang Z M,Wan S B,Ning T Y,et al.Effects of nitrogen level on nitrogen metabolism and correlating enzyme activity in peanut[J].Journal of Plant Ecology,2008,(6):1407-1416.(in Chinese)
[16]王月福,于振文,李尚霞,等.氮素营养水平对冬小麦氮代谢关键酶活性变化和籽粒蛋白质含量的影响[J].作物学报,2002,(6):743-748.Wang Y F,Yu Z W,Li S X,et al.Effect of nitrogen nutrition on the change of key enzyme activity during the nitrogen metabolism and kernel protein content in winter wheat[J].Crop Journal,2002,(6):743-748.(in Chinese)
[17]赵宏伟.不同氮素营养水平下春玉米碳氮代谢机理的研究[D].哈尔滨:东北农业大学,2003.Zhao H W.Study on the mechanism of carbon and nitrogen metabolism of spring maize under different nitrogen levels[D].Harbin:Northeast Agricultural University,2003.(in Chinese)
[18]岳红宾.不同氮素水平对烟草碳氮代谢关键酶活性的影响[J].中国烟草科学,2007,28(1):18-20.Yue H B.Effects of different nitrogen levels on the activities of key enzymes of carbon and nitrogen metabolism in tobacco[J].Chinese Tobacco Science,2007,28(1):18-20.(in Chinese)
[19]刘明,吕爱锋,武建军,等.干旱对农业生态系统影响研究进展[J].中国农学通报,2014,30(32):165-171.Liu M,Lv A F,Wu J J,et al.A review of impacts of drought on agro-ecosystem[J].Chinese Agricultural Science Bulletin,2014,30(32):165-171.(in Chinese)
[20]王奎稳.设施葡萄秋季施肥技术[J].落叶果树,2016,(3):52.Wang K W.Autumn fertilization techniques for protected grapes[J].Deciduous Fruit Tree,2016,(3):52.(in Chinese)
[21]倪荣梅.不同水肥管理对设施葡萄产量和品质的影响研究[J].新疆农垦科技,2016,39(4):62-64.Ni R M.Effects of different water and fertilizer management on yield and quality of protected grape[J].Xinjiang Agricultural Reclamation Science and Technology,2016,39(4):62-64.(in Chinese)
[22]Bao S D.Soil and agricultural chemistry analysis[M].Beijing:China Agriculture Press,1999.
[23]陈建勋,王晓峰.植物生理学实验指导[M].广州:华南理工大学出版社,2015.Chen J X,Wang X F.Experimental guidance of plant physiology[M].Guangzhou:South China University of Technology Press,2015.(in Chinese)
[24]叶利庭,吕华军,宋文静,等.不同氮效率水稻生育后期氮代谢酶活性的变化特征[J].土壤学报,2011,48(1):132-140.Ye L T,Lv H J,S W J,et al.Changes of nitrogen metabolism enzyme activities in late growth stage of rice with different nitrogen efficiency[J].Journal of Soil Science,2011,48(1):132-140.(in Chinese)
[25]向友珍,张富仓,范军亮,等.基于临界氮浓度模型的日光温室甜椒氮营养诊断[J].农业工程学报,2016,32(17):89-97.Xiang Y Z,Zhang F C,Fan J L,et al.Nutrition diagnosis for Nin bell pepper based on critical nitrogen model in solar greenhouse[J].Journal of the CSAE,2016,32(17):89-97.(in Chinese)
[26]孙园园,孙永健,杨志远,等.不同形态氮肥与结实期水分胁迫对水稻氮素利用及产量的影响[J].中国生态农业学报,2013,21(3):274-281.Sun Y Y,Sun Y J,Yang Z Y,et al.Effects of different forms of nitrogen fertilizer and water stress on nitrogen use and yield in rice[J].Chinese Journal of Ecological Agriculture,2013,21(3):274-281.(in Chinese)
[27]杨夕,郁松林,赵丰云,等.水分胁迫下硝酸钠对葡萄幼苗氮代谢酶活性的影响[J].北方园艺,2018,(12):27-35.Yang X,Yu S L,Zhao F Y,et al.Effect of sodium nitrate on the activity of nitrogen metabolism enzymes in grape seedlings under water stress[J].Northern Horticulture,2018,(12):27-35.(in Chinese)
[28]白泽晨.不同程度水分胁迫对赤霞珠葡萄幼树氮素吸收代谢的影响[D].石河子:石河子大学,2015.Ba Z C.Effects of water stress on nitrogen absorption and metabolism of cabernet sauvignon young trees[D].Shihezi:Shihezi University,2015.(in Chinese)
[29]周卫,马敬坤,王志强,等.渗水分和盐胁迫对水稻幼苗叶片氨同化的影响[J].武汉大学学报(理学版),2005,(4):521-524.Zhou W,Ma J K,Wang Z Q,et al.Effects of water seepage and salt stress on ammonia assimilation in rice seedling leaves[J].Journal of Wuhan University(Science Edition),2005,(4):521-524.(in Chinese)
[2]吉沐祥,李国平,芮东明,等.江苏省鲜食葡萄病虫害绿色防控技术规程[J].江苏农业科学,2015,43(7):107-109.Ji M X,Li G P,Rui D M,et al.Technical specification for green control of diseases and insect pests of fresh grapevine in Jiangsu province[J].Jiangsu Agricultural Science,2015,43(7):107-109.(in Chinese)
[3]张毅,石玉,胡晓辉,等.外源Spd对盐碱胁迫下番茄幼苗氮代谢及主要矿质元素含量的影响[J].应用生态学报,2013,24(5):1401-1408.Zhang Y,Shi Y,Hu X H,et al.Effects of exogenous Spd on nitrogen metabolism and contents of main mineral elements in tomato seedlings under salt and alkali stress[J].Journal of Applied Ecology,2013,24(5):1401-1408.(in Chinese)
[4]谷端银,王秀峰,高俊杰,等.纯化腐植酸对氮胁迫下黄瓜幼苗生长和氮代谢的影响[J].应用生态学报,2018,29(8):2575-2582.Gu D Y,Wang X F,Gao J J,et al.Effects of purified humic acid on the growth and nitrogen metabolism of cucumber seedlings under nitrogen stress[J].Journal of Applied Ecology,2018,29(8):2575-2582.(in Chinese)
[5]Andrews M,Raven JA,Lea PJ.Do plants need nitrate?The mechanisms by which nitrogen form affects plants[J].Annals of Applied Biology,2013,163:174-199.
[6]Gajewska E,Sklodowska M.Nickel-induced changes in nitrogen metabolism in wheat shoots[J].Journal of Plant Physiology,2009,166:1034-1044.
[7]Lam H M,Coschigano K T,Oliveira I C,et al.The moleculargenetics of nitrogen assimilation into amino acids in higher plants[J].Annual Review of Plant Biology,1996,47:569-593.
[8]Vega M,Marinod,Sanchez Z,et al.CO2 enrichment modulates ammonium nutrition in tomato adjusting carbon and nitrogen metabolism to stomatal conductance[J].Plant Sci,2015,41:32-44.
[9]丁邦琴,李铁军.谷氨酰胺高产菌株在不同NH4+浓度下的基因差异表达[J].基因组学与应用生物学,2013,32(1):41-45.Ding B Q,Li T J.Differential gene expression of high glutamine producing strain of arthrobact globiformis in the different NH4+concentration[J].Genomics and Applied Biology,2013,32(1):41-45.(in Chinese)
[10]Krapp A.Plant nitrogen assimilation and its regulation:a complex puzzle with missing pieces[J].Current Opinion in Plant Biology,2015,25:115-122.
[11]张华珍,徐恒玉.植物氮素同化过程中相关酶的研究进展[J].北方园艺,2011,(20):180-183.Zhang H Z,Xu H Y.Advances in the study of enzymes related to nitrogen assimilation in plants[J].Northern Horticulture,2011,(20):180-183.(in Chinese)
[12]孙永健,孙园园,蒋明金,等.施肥水平对不同氮效率水稻氮素利用特征及产量的影响[J].中国农业科学,2016,49(24):4745-4756.Sun Y J,Sun Y Y,Jiang M J,et al.Effects of fertilizer levels on nitrogen utilization characteristics and yield in rice cultivars with different nitrogen use efficiencies[J].Scientia Agricultura Sinica,2016,49(24):4745-4756.(in Chinese)
[13]于坤.滴灌方式和水氮处理对酿酒葡萄幼苗生理特性和根系形态的影响[D].石河子:石河子大学,2014.Yu K.Effects of irrigation methods and water and nitrogen application on root morphology and physiological characteristics of wine grapes seedlings[D].Shihezi:Shihezi University,2014.(in Chinese)
[14]张丽莹.水肥耦合对温室无土栽培水果黄瓜生长发育的影响及其生理机制的研究[D].呼和浩特:内蒙古农业大学,2011.Zhang L Y.Effects of water and fertilizer coupling on growth and development of fruit cucumber and its physiological mechanism under soilless culture in greenhouse[D].Hohhot:Agricultural University of the Inner Mongol,2011.(in Chinese)
[15]张智猛,万书波,宁堂原,等.氮素水平对花生氮素代谢及相关酶活性的影响[J].植物生态学报,2008,(6):1407-1416.Zhang Z M,Wan S B,Ning T Y,et al.Effects of nitrogen level on nitrogen metabolism and correlating enzyme activity in peanut[J].Journal of Plant Ecology,2008,(6):1407-1416.(in Chinese)
[16]王月福,于振文,李尚霞,等.氮素营养水平对冬小麦氮代谢关键酶活性变化和籽粒蛋白质含量的影响[J].作物学报,2002,(6):743-748.Wang Y F,Yu Z W,Li S X,et al.Effect of nitrogen nutrition on the change of key enzyme activity during the nitrogen metabolism and kernel protein content in winter wheat[J].Crop Journal,2002,(6):743-748.(in Chinese)
[17]赵宏伟.不同氮素营养水平下春玉米碳氮代谢机理的研究[D].哈尔滨:东北农业大学,2003.Zhao H W.Study on the mechanism of carbon and nitrogen metabolism of spring maize under different nitrogen levels[D].Harbin:Northeast Agricultural University,2003.(in Chinese)
[18]岳红宾.不同氮素水平对烟草碳氮代谢关键酶活性的影响[J].中国烟草科学,2007,28(1):18-20.Yue H B.Effects of different nitrogen levels on the activities of key enzymes of carbon and nitrogen metabolism in tobacco[J].Chinese Tobacco Science,2007,28(1):18-20.(in Chinese)
[19]刘明,吕爱锋,武建军,等.干旱对农业生态系统影响研究进展[J].中国农学通报,2014,30(32):165-171.Liu M,Lv A F,Wu J J,et al.A review of impacts of drought on agro-ecosystem[J].Chinese Agricultural Science Bulletin,2014,30(32):165-171.(in Chinese)
[20]王奎稳.设施葡萄秋季施肥技术[J].落叶果树,2016,(3):52.Wang K W.Autumn fertilization techniques for protected grapes[J].Deciduous Fruit Tree,2016,(3):52.(in Chinese)
[21]倪荣梅.不同水肥管理对设施葡萄产量和品质的影响研究[J].新疆农垦科技,2016,39(4):62-64.Ni R M.Effects of different water and fertilizer management on yield and quality of protected grape[J].Xinjiang Agricultural Reclamation Science and Technology,2016,39(4):62-64.(in Chinese)
[22]Bao S D.Soil and agricultural chemistry analysis[M].Beijing:China Agriculture Press,1999.
[23]陈建勋,王晓峰.植物生理学实验指导[M].广州:华南理工大学出版社,2015.Chen J X,Wang X F.Experimental guidance of plant physiology[M].Guangzhou:South China University of Technology Press,2015.(in Chinese)
[24]叶利庭,吕华军,宋文静,等.不同氮效率水稻生育后期氮代谢酶活性的变化特征[J].土壤学报,2011,48(1):132-140.Ye L T,Lv H J,S W J,et al.Changes of nitrogen metabolism enzyme activities in late growth stage of rice with different nitrogen efficiency[J].Journal of Soil Science,2011,48(1):132-140.(in Chinese)
[25]向友珍,张富仓,范军亮,等.基于临界氮浓度模型的日光温室甜椒氮营养诊断[J].农业工程学报,2016,32(17):89-97.Xiang Y Z,Zhang F C,Fan J L,et al.Nutrition diagnosis for Nin bell pepper based on critical nitrogen model in solar greenhouse[J].Journal of the CSAE,2016,32(17):89-97.(in Chinese)
[26]孙园园,孙永健,杨志远,等.不同形态氮肥与结实期水分胁迫对水稻氮素利用及产量的影响[J].中国生态农业学报,2013,21(3):274-281.Sun Y Y,Sun Y J,Yang Z Y,et al.Effects of different forms of nitrogen fertilizer and water stress on nitrogen use and yield in rice[J].Chinese Journal of Ecological Agriculture,2013,21(3):274-281.(in Chinese)
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