不同酸度欧李果实有机酸积累特性与相关代谢酶活性分析

doi:10.3864/j.issn.0578-1752.2013.19.016

摘要/Abstract

摘要： 【目的】比较不同欧李品种果实中主要有机酸积累特性及相关酶活性。【方法】以6年生欧李品种‘农大3号’和‘农大4号’为材料，采用超高效液相色谱（UPLC）检测果实发育过程中总酸、苹果酸和柠檬酸含量，并测定苹果酸和柠檬酸相关酶活性。【结果】两个品种果实中苹果酸和柠檬酸主要在果实发育后期积累，品种间积累速率存在较大差异。果实发育前期NADP-苹果酸酶（NADP-ME）活性较高，NAD-苹果酸脱氢酶（NAD-MDH）活性和磷酸烯醇式丙酮酸羧化酶（PEPC）活性相对较低，使苹果酸仅少量积累；果实发育后期NADP-ME活性迅速下降，而NAD-MDH活性和PEPC活性开始上升，促进了苹果酸大量积累；果实成熟时NADP-ME活性突然升高，而NAD-MDH活性和PEPC活性开始下降，促使苹果酸降解。果实成熟前（花后18-19周）‘农大4号’NAD-MDH活性显著高于‘农大3号’、而NADP-ME活性低于‘农大3号’，导致成熟时前者苹果酸含量高于后者。花后17-19周‘农大3号’柠檬酸合成酶（CS）活性高于‘农大4号’，成熟时柠檬酸含量也高于后者。【结论】果实发育后期是苹果酸和柠檬酸积累的关键时期，苹果酸的积累差异主要由NAD-MDH活性和NADP-ME活性协同变化引起，柠檬酸积累差异主要受CS活性变化影响。

关键词: 欧李 , 果实 , 苹果酸 , 柠檬酸 , 代谢酶

Abstract: 【Objective】 The accumulation characteristics of main organic acid and activities of acid-related enzymes between two cultivars of Chinese dwarf cherry with different acidities were compared. 【Method】 Using the method of ultra performance liquid chromatograph (UPLC), the contents of total organic acids, malic acid and citric acid, as well as the activities of organic acid-metabolizing enzymes were analyzed during the fruit development of 6-year-old Chinese dwarf cherry ‘Nongda 3’ and ‘Nongda 4’. 【Result】Malic acid and citric acid of these two varieties accumulated mainly in the late period of fruit development, but their accumulation rates were quite different. Because of the high NADP-ME activity and the low NAD-MDH and PEPC activities, there was only a small amount of malic acid accumulation in the early period of fruit development. During the late period, NADP-ME activity decreased rapidly and NAD-MDH activity and PEPC activity began increasing, which promoted a massive accumulation of malic acid. The sudden increase of NADP-ME activity and the decline of NAD-MDH activity and PEPC activity caused the degradation of mailc acid at the ripening stage. Before the ripening stage(18-19 weeks after anthesis), NAD-MDH activity of ‘Nongda 4’ was higher than the activity of ‘Nongda 3’ , while NADP-ME activity of ‘Nongda 4’ was lower. As a result, the content of malic acid of ‘Nongda 4’ was higher than the content of ‘Nongda 3’. The content of citric acid of ‘Nongda 3’ was higher (17-19 weeks after anthesis) because its CS activity was higher at the ripening stage.【Conclusion】The late period of fruit development is a critical period for the accumulation of malic acid and citric acid. The difference in malic acid accumulation is mainly caused by the coordination change of NAD-MDH activity and NADP-ME activity, while the difference in the accumulation of citric acid is mainly affected by the change of CS activity.

Key words: Chinese dwarf cherry , fruit , malic acid , citric acid , metabolism-related enzymes

王鹏飞1, 薛晓芳12, 穆晓鹏1, 张建成1, 曹琴1, 杜俊杰1. 不同酸度欧李果实有机酸积累特性与相关代谢酶活性分析[J]. 中国农业科学, 2013, 46(19): 4101-4109.

WANG Peng-Fei-1, XUE Xiao-Fang-12, MU Xiao-Peng-1, ZHANG Jian-Cheng-1, CAO Qin-1, DU Jun-Jie-1. Analysis of Organic Acid Accumulation Characteristics and Organic Acid-Metabolizing Enzyme Activities of Chinese Dwarf Cherry (Cerasus humilis Bunge) Fruit[J]. Scientia Agricultura Sinica, 2013, 46(19): 4101-4109.

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参考文献

[1]张上隆, 陈昆松. 果实品质形成与调控的分子生理. 北京: 中国农业出版社, 2007: 67-80.

Zhang S L, Chen K S. Molecular Physiology of Fruit Quality Development and Regulation. Beijing: China Agriculture Press, 2007: 67-80. (in Chinese)

[2]王鹏飞, 曹琴, 何永波, 杜俊杰. 欧李果实发育期糖和酸组分及其含量的动态变化特性. 西北植物学报, 2011, 31(7): 1411-1416.

Wang P F, Cao Q, He Y B, Du J J. Dynamic changes of sugars and acids of fruit development period in Chinese dwarf cherry. Acta Botanica Boreali-Occidentalia Sinica, 2011, 31(7): 1411-1416. (in Chinese)

[3]陈发兴, 刘星辉, 陈立松. 果实有机酸代谢研究进展. 果树学报, 2005, 22(5): 526-531.

Chen F X, Liu X H, Chen L S. Advance in research on organic acid metabolism in fruits. Journal of Fruit Science, 2005, 22(5): 526-531.（in Chinese）

[4]李庆余, 徐新娟, 朱毅勇, 董彩霞, 沈其荣. 半定量RT- PCR研究氮素形态对樱桃番茄果实中有机酸代谢相关酶基因表达的影响. 植物营养与肥料学报, 2011, 17(2): 341-348.

Li Q Y, Xu X J, Zhu Y Y, Dong C X, Shen Q R. Organic acid metabolism and related genes expression in cherry tomato fruit under different nitrogen fertilization using semi quantitative RT- PCR. Plant Nutrition and Fertilizer Science, 2011, 17(2): 341- 348. (in Chinese)

[5]姚玉新, 李明, 由春香, 刘志, 王冬梅, 郝玉金. 苹果果实中苹果酸代谢关键酶与苹果酸和可溶性糖积累的关系. 园艺学报, 2010, 37(1): 1-8.

Yao Y X, Li M, You C X, Liu Z, Wang D M, Hao Y J. Relationship between malic acid metabolism related key enzymes and accumulation of malic acid as well as the soluble sugars in apple fruit. Acta Horticulturae sinica, 2010, 37(1): 1-8. (in Chinese)

[6]赵永红, 李宪利, 姜泽盛, 王长健, 杨富林. 设施油桃果实发育过程中有机酸酸代谢的研究. 中国生态农业学报, 2007, 15(5): 87-89.

Zhao Y H, Li X L, Jiang Z S, Wang C J, Yang F L. Organic acid metabolism in nectarine fruit development under protected cultivation. Chinese Journal of Eco-Agriculture, 2007, 15(5): 87-89. (in Chinese)

[7]文涛, 熊庆娥, 曾伟光, 刘远鹏. 脐橙果实发育过程中有机酸代谢合成酶活性的变化. 园艺学报, 2001, 28(2): 161-163.

Wen T, Xiong Q E, Zeng W G, Liu Y P. Changes of organic acid synthetase activity during fruit development of Navel orange ( Citrus sinesis Osbeck). Acta Horticulturae Sinica, 2001, 28(2): 161-163. (in Chinese)

[8]陈美霞, 赵从凯, 陈学森, 郝会军, 张宪省. 杏果实发育过程中有机酸积累与相关代谢酶的关系.果树学报, 2009, 26(4): 471-474.

Chen M X, Zhao C K, Chen X S, Hao H J, Zhang X S. Relationship between accumulation of organic acid and organic acid-metabolizing enzymes during apricot fruit development. Journal of Fruit Science, 2009, 26(4): 471-474. (in Chinese)

[9]Sadka A, Dahan E, Or E, Roose M L, Marsh K B, Cohen L. Comparative analysis of mitochondrial citrate synthase gene structure, transcript level and enzymatic activity in acidless and acid-containing Citrus varieties. Australian Journal of Plant Physiology, 2001, 28(5): 383-390.

[10]Canel C, Bailey-Serres J N, Roose M L. Molecular characterization of the mitochondrial citrate syntheses gene of an acidless pummelo (Citrus maxima). Plant Molecular Biology, 1996, 31(15): 143-147.

[11]Saradhuldhat P, Paull R E. Pineapple organic acid metabolism and accumulation during fruit development. Scientia Horticulturae, 2007, 112(3): 297-303.

[12]韩振海, 陈昆松. 实验园艺学. 北京: 高等教育出版社, 2006: 444-446.

Han Z H, Chen K S. Experimental Horticulture. Beijing: Higher Education Press, 2006: 444-446. (in Chinese)

[13]Hirai M, Ueno I. Development of citrus fruits: fruit development and enzymatic changes in juice vesicle tissue. Plant and Cell Physiology, 1977, 18(4): 791-799.

[14]Sadka A, Dahan E, Cohen L, Marsh K B. Aconitase activity and expression during the development of lemon fruit. Physiologia Plantarum, 2000, 108(3): 255-262.

[15]Chen M, Jiang Q, Yin X R, Lin Q, Chen J Y, Allan A C, Xu C J, Chen K S. Effect of hot air treatment on organic acid and sugar-metabolism in Ponkan (Citrus reticulata) fruit. Scientia Horticulturae, 2012, 147: 118-125.

[16]Xu X J, Li Q Y, Song X H, Shen Q R, Dong C X. Dynamic regulation of nitrogen and organic acid metabolism of cherry tomato fruit as affected by different nitrogen forms. Pedosphere, 2012, 22(1): 67-78.

[17]Thakur A, Singh Z. Responses of ‘Spring Bright’ and ‘Summer Bright’nectarines to deficit irrigation: Fruit growth and concentration of sugars and organic acids. Scientia Horticulturae, 2012, 135: 112-119.

[18]Sweetman C, Deluc L G, Cramer G. R, Ford C M, Soole K L. Regulation of malate metabolism in grape berry and other developing fruits. Phytochemistry, 2009, 70: 1329-1344.

[19]Yao Y X, Li M, Liu Z, You C X, Wang D M, Zhai H, Hao Y J. Molecular cloning of three malic acid related genes MdPEPC, MdVHA-A, MdcyME and their expression analysis in apple fruits. Scientia Horticulturae, 2009, 122(3): 404-408.

[20]Yao Y X, Li M, Zhai H, You C X, Hao Y J. Isolation and characterization of an apple cytosolic malate dehydrogenase gene reveal its function in malate synthesis. Journal of Plant Physiology, 2011, 168(5): 474-480.

[21]Etienne C, Moing A, Dirlewanger E. Raymond P, Monet R, Rothan C. Isolation and characterization of six peach cDNAs encoding key proteins in organic acid metabolism and solute accumulation involvement in regulating peach fruit acidity. Physiologial Plantarum, 2002, 114(2): 259-270.

[22]Edwards G E, Andreo CS. NADP-malic enzyme from plants. Phytochemistry, 1992, 31(6): 1845 -1857.

[23]Moing A, Svanella L, Gaudillere M, Gaudillere J P, Monet R. Organic acid concentration is little controlled by phosphoenolpyruvate carboxylase activity in peach fruit. Australian Journal of Plant Physiology, 1999, 26(6): 579-585.

[24]Diakou P, Svanella L, Raymond P, Gaudillere J P, Moing A. Phosphoenolpyruvate carboxylase during grape berry development: protein level, enzyme activity and regulation. Australian Journal of Plant Physiology, 2000, 27(3): 221-229.

[25]Kubo T, Kihara T, Hirabayashi T. The effect of spraying lead arsenate on citrate accumulation and the related enzyme activities in the juice saces of Citrus natsudaidai. Journal of the Japanese Society for Horticultural Science, 2002, 71(3): 305-310.

[26]罗安才, 杨晓红, 邓英毅, 李纯凡, 向可术, 李道高. 柑橘果实发育过程中有机酸含量及相关代谢酶活性的变化.中国农业科学, 2003, 36(8): 941-944.

Luo A C, Yang X H, Deng Y Y, Li C F, Xing K S, Li D G. Organic acid Concentrations and the relative enzymatic changes during the development of the citrus fruits. Scientia Agricultura Sinica, 2003, 36(8): 941-944. (in Chinese)