甜瓜幼苗叶片内源一氧化氮和蔗糖代谢对低温胁迫的响应
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摘要
以低温敏感型甜瓜品种‘XL-1’和耐低温型品种‘红优’为试材,采用6℃低温处理0、1、3、6、12、24h及3d、5d和7d,研究低温胁迫下甜瓜幼苗叶片中NO合成和蔗糖代谢的变化特征。结果表明:(1)低温胁迫能提高甜瓜幼苗叶片中硝酸还原酶(NR)活性,诱导促进NO生成,其中耐低温型甜瓜‘红优’中NO对低温的响应时间更早,变化幅度更大。(2)与对照相比,低温胁迫处理提高了2种甜瓜幼苗叶片中蔗糖、果糖和葡萄糖含量,增加了蔗糖磷酸合成酶(SPS)、蔗糖合成酶(SS)、酸性转化酶(AI)和中性转化酶(NI)活性,降低了淀粉含量。(3)低温胁迫处理使2种甜瓜叶片中渗透调节物质可溶性糖、可溶性蛋白、脯氨酸的含量上升。研究发现,低温胁迫通过增加甜瓜体内NO合成酶的活性刺激体内NO合成,通过促进蔗糖代谢相关酶的活性,提高蔗糖、果糖和葡萄糖的含量,从而响应低温胁迫,且低温胁迫诱导的糖分物质积累时间晚于NO的产生时间。
In order to investigate the changes of NO synthesis and sucrose metabolism in melon seedlings under chilling stress,cultivar‘Hongyou'(chilling-tolerant)and ‘XL-1'(chilling-sensitive)melon seedlings were treated with 6℃ for 0,1,3,6,12,24 h,3 d,5 dand 7 din climate chambers.The results showed that:(1)chilling stress increased the activity of nitrate reductase(NR)and induced NO production.The response time and change of NO to chilling stress were earlier and more obvious in chilling-tolerant cultivar‘Hongyou'.(2)Compare with the control,the chilling stress condition increased sucrose,fructose and glucose contents,enhanced the activities of sucrose phosphate synthase(SPS),sucrose synthase(SS),acid invertase(AI),neutral invertase(NI),and decreased starch content.(3)The contents of soluble sugar,soluble protein and proline in the leaves of the two melon seedlings increased under chilling stress.These results indicated that melon seedlings could respond to chilling stress by inducing NO and carbohydrate generation.In addition,the accumulation of sugar substances induced by chilling stress was later than the production of NO.
In order to investigate the changes of NO synthesis and sucrose metabolism in melon seedlings under chilling stress,cultivar‘Hongyou'(chilling-tolerant)and ‘XL-1'(chilling-sensitive)melon seedlings were treated with 6℃ for 0,1,3,6,12,24 h,3 d,5 dand 7 din climate chambers.The results showed that:(1)chilling stress increased the activity of nitrate reductase(NR)and induced NO production.The response time and change of NO to chilling stress were earlier and more obvious in chilling-tolerant cultivar‘Hongyou'.(2)Compare with the control,the chilling stress condition increased sucrose,fructose and glucose contents,enhanced the activities of sucrose phosphate synthase(SPS),sucrose synthase(SS),acid invertase(AI),neutral invertase(NI),and decreased starch content.(3)The contents of soluble sugar,soluble protein and proline in the leaves of the two melon seedlings increased under chilling stress.These results indicated that melon seedlings could respond to chilling stress by inducing NO and carbohydrate generation.In addition,the accumulation of sugar substances induced by chilling stress was later than the production of NO.
引文
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[29]王燕凌,廖康,刘君,等.越冬前低温锻炼期间不同品种葡萄枝条中渗透性物质和保护酶活性的变化[J].果树学报,2006,23(3):375-378.WANG Y L,LIAO K,LIU J,et al.Changes of contents of osmosis substances and antioxidant enzyme activity in grape vines during cold exercise period before over-wintering[J].Journal of Fruit Science,2006,23(3):375-378.
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[36]KLOTZ K L,HAAGENSON D M.Wounding,anoxia and cold induce sugar beet sucrose synthase transcriptional changes that are unrelated to protein expression and activity[J].Journal of Plant Physiology,2008,165(4):423-434.
[37]石永春,刘卫群,王小彦.低温对烟草叶片中蔗糖向顶端分生组织转运的影响[J].中国农业科学,2009,42(12):4 390-4 395.SHI Y C,LIU W Q,WANG X Y.Effect of low temperature on sucrose transportation from leaves to apical meristem in tobacco[J].Scientia Agricultura Sinica,2009,42(12):4 390-4 395.
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[2]DALEN L S,JOHNSEN,LNNEBORG A,et al.Freezing tolerance in Norway spruce,the potential role of pathogenesis-related proteins[J].Acta Physiol.Plant,2015,37(1):1-9.
[3]BALIBREA M E,DELL' AMICO J,BOLARIN M C,et al.Carbon partitioning and sucrose metabolism in tomato plants growing under salinity[J].Physiol.Plant,2000,110(4):503-511.
[4]SIDDIQUI M H,Al-WHAIBI M H,BASALAH M O.Role of nitric oxide in tolerance of plants to abiotic stress[J].Protoplasma,2011,248(3):447-455.
[5]YU M,LAMATTINA L,SPOEL S H,et al.Nitric oxide function in plant biology:a redox cue in deconvolution[J].New Phytol.,2014,202(4):1 142-1 156.
[6]LASPINA,N V,GROPPA M D,TOMARO M L,et al.Nitric oxide protects sunflower leaves against Cd-induced oxidative stress[J].Plant Sci.,2005,169(2):323-330.
[7]SHI H,YE T,ZHU J K,et al.Constitutive production of nitric oxide leads to enhanced drought stress resistance and extensive transcriptional reprogramming in Arabidopsis[J].Journal of Experimental Botany,2014,65(15):4 119-4 131.
[8]LAMATTINA L,GARCIA-MATA C,GRAZIANO M,et al.Nitric oxide:the versatility of an extensive signal molecule[J].Annu.Rev.Plant Biol.,2003,54(1):109-136.
[9]ZHAO M G,CHEN L,ZHANG L L,et al.Nitric reductasedependent nitric oxide production is involved in cold acclimation and freezing tolerance in Arabidopsis[J].Plant Physiology,2009,151(2):755-767.
[10]SEHRAWATA,ABAT J K,DESWAL R.Rubisco depletion improved proteome coverage of cold responsive S-nitrosylated targets in Brassica juncea[J].Frontiers in Plant Science,2013,4(342):342.
[11]SONG L,DING W,ZHAO M,et al.Nitric oxide protects against oxidative stress under heat stress in the calluses from two ecotypes of reed[J].Plant Science,2006,171(4):449-458.
[12]GOULD K S,LAMOTTE O,KLINGUER A,et al.Nitric oxide production in tobacco leaf cells:ageneralized stress response[J].Plant,Cell&Environment,2003,26(11):1 851-1 862.
[13]AIRAKI M,LETERRIER M,MATEOS R M,et al.Metabolism of reactive oxygen species and reactive nitrogen species in pepper(Capsicum annuum L.)plants under low temperature stress[J].Plant,Cell&Environment,2012,35(2):281-295.
[14]PUYAUBERT J,BAUDOUIN E.New clues for a cold case:nitric oxide response to low temperature[J].Plant,Cell&Environment,2014,37(12):2 623-2 630.
[15]姚远,闵义,胡新文,等.低温胁迫对木薯幼苗叶片转化酶及可溶性糖含量的影响[J].热带作物学报,2010,31(4):556-560.YAO Y,MIN Y,HU X W,et al.Effects of low temperature stress on the activity of invertase and soluble sugar content in leaves of cassava seedlings[J].Chinese Journal of Tropical Crops,2010,31(4):556-560.
[16]齐红岩,姜岩岩,华利静.短期夜间低温对栽培番茄和野生番茄果实蔗糖代谢的影响[J].园艺学报,2012,39(2):281-288.QI H Y,JIANG Y Y,HUA L J.Responses to short-term low night temperature of sucrose-metabolizing in domesticated tomato and wild relative[J].Acta Horticulturae Sinica,2012,39(2):281-288.
[17]SCHEIBLE W R,GONZALEZ-FONTES A,LAUERER M,et al.Nitrate acts as a signal to induce organic acid metabolism and repress starch metabolism in tobacco[J].The Plant Cell Online,1997,9(5):783-798.
[18]HUANG C H,YU B,TENG Y W,et al.Effects of fruit bagging on coloring and related physiology,and qualities of red Chinese sand pears during fruit maturation[J].Scientia Horticulturae,2009,121(2):149-158.
[19]齐红岩,李天来,张洁,等.番茄果实发育过程中糖的变化与相关酶活性的关系[J].园艺学报,2006,33(2):294-299.QI H Y,LI T L,ZHANG J,et al.Relationship between carbohydrate change and related enzymes activities during tomato fruit development[J].Acta Horticulturae Sinica,2006,33(2):294-299.
[20]BALIBREA M E,RUS-ALVAREZ A M,BOLARIN M C.,et al.Fast changes in soluble carbohydrates and proline contents in tomato seedlings in response to ionic and non-ionic iso-osmotic stress[J].Journal of Plant Physiology,1997,151(2):221-226.
[21]张殿忠,汪沛洪,赵会贤.测定小麦叶片游离脯氨酸含量的方法[J].植物生理学通讯,1990,(4):62-65.ZHANG D Z,WANG P H,ZHAO H X.Determination of the content of free proline in wheat leaves[J].Plant Physiology Communications,1990,(4):62-65.
[22]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,2000,167-169.
[23]HEATH R L,PACKER L.Photoperoxidation in isolated chloroplasts:I.Kinetics and stoichiometry of fatted acid peroxidation[J].Arch.Biochem.Biophys,1968,125(1):189-198.
[24]ZHANG S,JIANG H,PENG S,et al.Sex-related differences in morphological,physiological,and ultrastructural responses of Populus cathayana to chilling[J].Journal of Experimental Botany,2010,62(2):675-686.
[25]胡俊杰,张古文,胡齐赞,等.低温胁迫对菜用大豆生长、叶片活性氧及多胺代谢的影响[J].浙江农业学报,2011,23(6):1 113-1 118.HU J J,ZHANG G W,HU Q Z,et al.Effects of chilling stress on growth,metabolism of reactive oxygen species and polyamines in vegetable soybean seedlings[J].Acta Agriculturae Zhejiangensis,2011,23(6):1 113-1 118.
[26]THEOCHARIS A,CLEMENT C,BARKA E A.Physiological and molecular changes in plants grown at low temperatures[J].Planta,2012,235(6):1 091-1 105.
[27]MAJLATH I,SZALAI G,SOOS V,et al.Effect of light on the gene expression and hormonal status of winter and spring wheat plants during cold hardening[J].Physiologia Plantarum,2012,145(2):296-314.
[28]BELIGNI M V,LAMATTINA L.Nitric oxide in plants:the history is just beginning[J].Plant,Cell&Environment,2001,24(3):267-278.
[29]王燕凌,廖康,刘君,等.越冬前低温锻炼期间不同品种葡萄枝条中渗透性物质和保护酶活性的变化[J].果树学报,2006,23(3):375-378.WANG Y L,LIAO K,LIU J,et al.Changes of contents of osmosis substances and antioxidant enzyme activity in grape vines during cold exercise period before over-wintering[J].Journal of Fruit Science,2006,23(3):375-378.
[30]岳海,李国华,李国伟,等.澳洲坚果不同品种耐寒特性的研究[J].园艺学报,2010,37(1):31-38.YUE H,LI G H,LI G W,et al.Studies on cold resistance of different macadamia cultivars[J].Acta Horticulturae Sinica,2010,37(1):31-38.
[31]LIU L J,CANG J,YU J,et al.Effects of exogenous abscisic acid on carbohydrate metabolism and the expression levels of correlative key enzymes in winter wheat under low temperature[J].Biosci.Biotechnol.Biochem.,2013,77(3):516-525.
[32]WONG B L,BAGGETT K L,RYE A H.Seasonal patterns of reserve and soluble carbohydrates in mature sugar maple(Acer saccharum)[J].Canadian Journal of Botany,2003,81(8):780-788.
[33]KEREPESI I,BANYAI-STEFANOVITS,GALIBA G.Cold acclimation and abscisic acid induced alterations in carbohydrate content in calli of wheat genotypes differing in frost tolerance[J].Journal of Plant Physiology,2004,161(1):131.
[34]SASAKA H,ICHIMURA K,IMADA S,et al.Sucrose synthase and sucrose phosphate synthase,but not acid invertase,are regulated by cold acclimation and deacclimation in cabbage seedling[J].Journal of Plant Physiology,2001,158(7):847-852.
[35]郑春芳,刘伟成,陈少波,等.短期夜间低温胁迫对秋茄幼苗碳氮代谢及其相关酶活性的影响[J].生态学报,2013,33(21):6 853-6 862.ZHENG C F,LIU W C,CHEN S B,et al.Effects of shortterm dark chilling on leaves carbon and nitrogen metabolism and involved activities of enzymes in mangrove Kandelia obovata seedling[J].Acta Ecologica Sinica,2013,33(21):6853-6 862.
[36]KLOTZ K L,HAAGENSON D M.Wounding,anoxia and cold induce sugar beet sucrose synthase transcriptional changes that are unrelated to protein expression and activity[J].Journal of Plant Physiology,2008,165(4):423-434.
[37]石永春,刘卫群,王小彦.低温对烟草叶片中蔗糖向顶端分生组织转运的影响[J].中国农业科学,2009,42(12):4 390-4 395.SHI Y C,LIU W Q,WANG X Y.Effect of low temperature on sucrose transportation from leaves to apical meristem in tobacco[J].Scientia Agricultura Sinica,2009,42(12):4 390-4 395.
[38]胡瑞芳,姜慧,李玥莹.蔗糖代谢相关酶的研究进展[J].北方园艺,2012,(1):167-170.HU R F,JIANG H,LI Y Y.Research advance on sucrose synthesize enzymes[J].Northern Horticulture,2012,(1):167-170.
[39]牛俊奇,檀小辉,黄静丽,等.低温胁迫对不同耐寒型甘蔗蔗糖代谢相关酶基因表达及其酶活性的影响[J].南方农业学报,2014,45(9):1 566-1 573.NIU J Q,TAN X H,HUANG J L,et al.Effect of chilling stress on sucrose metabolism enzymes in leaf of sugarcane varieties with different cold resistance[J].Journal of Southern Agriculture,2014,45(9):1 566-1 573.
[40]SHEEN J,ZHOU L,JANG J C.Sugars as signaling molecules[J].Curr.Opin.Plant Biol.,1999,2(5):410-418.
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