潘德灼;钟灿钰;邓朝军;卢思;蒋际谋;陈伟;

• 1:福建农林大学生命科学学院
• 2:福建省农业科学院果树研究所
• 3:福建农林大学林学院

摘要(Abstract)：

【目的】研究枇杷果实成熟过程中不同发育阶段(青果期、转色期和成熟期)的生理代谢变化及果实品质形成机制。【方法】以‘解放钟’枇杷品种为试验材料,采用蛋白质双向凝胶电泳及MALDI-TOF/TOF-MS技术分离并鉴定蛋白质,采用RT-qPCR技术分析基因表达水平变化,同时应用生理生化技术分析生理代谢变化。【结果】成功鉴定出55个差异表达蛋白,这些蛋白主要参与糖代谢(16%)、三羧酸循环与能量代谢(9%)、抗坏血酸-谷胱甘肽循环(14%)、类胡萝卜素合成及乙烯合成(11%)、蛋白质合成(13%)、细胞结构(8%)、防御与胁迫(9%)以及其它调控(13%)等生物学过程。随着果实成熟,参与糖代谢、三羧酸循环与能量代谢以及类胡萝卜素合成及乙烯合成过程的相关蛋白均上调表达;RT-qPCR的结果与蛋白质丰度变化趋势相一致;同时,糖类物质、类胡萝卜素以及维生素C等的含量不断积累,有机酸含量不断减少。【结论】在蛋白质丰度、生理生化水平以及mRNA水平上的结果共同表明糖代谢、三羧酸循环及能量代谢、类胡萝卜素合成以及乙烯合成等代谢途径在枇杷果实品质形成过程中起着重要作用。这些结果为枇杷果实品质改良提供重要的理论依据,并为今后枇杷新品种的培育奠定分子生物学基础。

关键词(KeyWords)： 枇杷;发育时期;蛋白质组;品质形成

Abstract:

Keywords:

基金项目(Foundation): 国家农作物种质资源共享服务平台(NICGR2018-054);; 福建省农业科学院科技创新团队建设项目(STIT2017-1-4);; 福建省自然科学基金项目(2014J01099)

作者(Author): 潘德灼;钟灿钰;邓朝军;卢思;蒋际谋;陈伟;

Email:

参考文献(References)：

• [1]王博,朴一龙,王琳,朴日子.野生软枣猕猴桃果实生长发育过程中生理生化变化[J].延边大学农学学报,2011,33(1):6-9.WANG Bo,PIAO Yilong,WANG Lin,PIAO Rizi. Changes of physio-biochemistry during the growth and development in undomesticated Actinidia arguta fruits[J]. Journal of Agricultural Science Yanbian University,2011,33(1):6-9.
• [2]魏建梅,齐秀东,朱向秋,马锋旺.苹果果实糖积累特性与品质形成的关系[J].西北植物学报,2009,29(6):1193-1199.WEI Jianmei,QI Xiudong,ZHU Xiangqiu,MA Fengwang. Relationship between the characteristics of sugar accumulation and fruit quality in apple(Malus domestica Borkh.)fruit[J]. Acta Botanica Borealia-Occidentalia Sinica,2009,29(6):1193-1199.
• [3]沈志军,马瑞娟,俞明亮,蔡志翔,宋宏峰,李晓.桃果实发育过程中主要糖及有机酸含量的变化分析[J].华北农学报,2007,22(6):130-134.SHEN Zhijun,MA Ruijuan,YU Mingliang,CAI Zhixiang,SONG Hongfeng,LI Xiao. Regularity analysis of main sugar and acid in fruit development of peach[J]. Acta Agriculturae Boreali-Sinica,2007,22(6):130-134.
• [4]王涛,谢小波,蔡美艳,黄雪燕,陈丹霞,戚行江.杨梅新品种黑晶的果实发育与主要品质指标变化规律[J].浙江农业学报,2007,19(6):427-430.WANG Tao,XIE Xiaobo,CAI Meiyan,HUANG Xueyan,CHEN Danxia,QI Xingjiang. Characteristics of fruit development and changes in main quality components in the fruits for new cultivar(cv. Heijing)of red bayberry[J]. Acta Agriculturae Zhejiangensis,2007,19(6):427-430.
• [5]王惠聪,黄辉白,黄旭明.荔枝果实的糖积累与相关酶活性[J].园艺学报,2003,30(1):1-5.WANG Huicong,HUANG Huibai,HUANG Xuming. Sugar accumulation and related enzyme activities in the litchi fruit of‘Nuomici’and‘Feizixiao’[J]. Acta Horticulturae Sinica,2003,30(1):1-5.
• [6]乔永旭,刘栓桃,赵智中,邢国明,何启伟.甜瓜果实发育过程中糖积累与蔗糖代谢相关酶的关系[J].果树学报,2004,21(5):447-450.QIAO Yongxu,LIU Shuantao,ZHAO Zhizhong,XING Guoming,HE Qiwei. Study on the correlation of sugar accumulation and sucrose-metabolizing enzymes during the development period of muskmelon(Cucumis melon)[J]. Journal of Fruit Science,2004,21(5):447-450.
• [7]赵智中,张上隆,徐昌杰,陈昆松,刘拴桃.蔗糖代谢相关酶在温州蜜柑果实糖积累中的作用[J].园艺学报,2001,28(2):112-118.ZHAO Zhizhong,ZHANG Shanglong,XU Changjie,CHEN Kunsong,LIU Shuantao. Roles of sucrose-metabolizing enzymes in accumulation of sugars in satsuma mandarin fruit[J].Acta Horticulturae Sinica,2001,28(2):112-118.
• [8] SOULEYRE E J F,IANNETTA P P M,ROSS H A,HANCOCK R D,SHEPHERD L V T,VIOLA M,TAYLOR M A,DAVIES H V. Starch metabolism in developing strawberry(Fragaria×ananassa)fruits[J]. Physiologia Plantarum,2004,121(3):369-376.
• [9]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[J].果树学报,2009,26(4):471-474.CHEN Meixia,ZHAO Congkai,CHEN Xuesen,HAO Huijun,ZHANG Xiansheng. Relationship between accumulation of organic acid and organic acid-metabolizing enzymes during apricot fruit development[J]. Journal of Fruit Science,2009,26(4):471-474.
• [10]龚荣高,吕秀兰,张光伦,胡世学.脐橙果实发育过程中有机酸代谢相关酶的研究[J].四川农业大学学报,2006,24(4):402-404.GONG Ronggao,LǜXiulan,ZHANG Guanglun,HU Shixue.Study on the organic acid-metabolizing enzymes in robertson navel orange during the fruit development[J]. Journal of Sichuan Agricultural University,2006,24(4):402-404.
• [11]于洋,翁倩,周宝利.番茄果实成熟过程中不同部位类胡萝卜素和糖含量变化及其相关性[J].沈阳农业大学学报,2012,42(6):683-687.YU Yang,WENG Qian,ZHOU Baoli. Changes in contents of carotenoids and sugar in peparate tissues and their correlation during ripening of tomato fruit[J]. Journal of Shenyang Agricultural University,2011,42(6):683-687.
• [12]李靖,孙淑霞,陈栋,涂美艳,刘佳,江国良.枇杷白肉突变体与野生型生物学特性和果实品质比较分析[J].西南农业学报,2017,30(7):1495-1498.LI Jing,SUN Shuxia,CHEN Dong,TU Meiyan,LIU Jia,JIANG Guoliang. Comparative analysis of biological characteristics and fruit quality in white-flesh loquat mutant and its wild type[J]. Southwest China Journal of Agricultural Sciences,2017,30(7):1495-1498.
• [13] JIANG J M,LIN Y X,CHEN Y Y,DENG C J,GONG H W,XU Q Z,ZHENG S Q,CHEN W. Proteomics approach reveals mechanism underlying susceptibility of loquat fruit to sunburn during color changing period[J]. Food Chemistry,2015,176:388-395.
• [14]张玲,郑婷婷,魏伟淋,朱云美,高用顺,杨向晖,林顺权.两种不同肉色枇杷果实类胡萝卜素积累及合成相关基因的表达[J].园艺学报,2015,42(11):2153-2162.ZHANG Ling,ZHENG Tingting,WEI Weilin,ZHU Yunmei,GAO Yongshun,YANG Xianghui,LIN Shunquan. Characterization of carotenoids accumulation and carotenogenic genes expression during fruit development in two loquat cultivars with different colour[J]. Acta Horticulturae Sinica,2015,42(11):2153-2162.
• [15] HADJIPIERI M,GEORGIADOU E C,MARIN A,DIAZ-MULA H M,GOULAS V,FOTOPOULOS V,TOMáS-BARBERáN F A,MANGANARIS G A. Metabolic and transcriptional elucidation of the carotenoid biosynthesis pathway in peel and flesh tissue of loquat fruit during on-tree development[J]. BMC Plant Biology,2017,17(1):102.
• [16]蒋际谋,邓朝军,林永祥,龚慧文,许奇志,郑少泉,陈伟.枇杷果皮响应高温强光胁迫的蛋白质组分析[J].热带亚热带植物学报,2014,22(4):383-390.JIANG Jimou,DENG Chaojun,LIN Yongxiang,GONG Huiwen,XU Qizhi,ZHENG Shaoquan,CHEN Wei. Proteomic analysis of loquat peels under high temperature and strong light stresses[J]. Journal of Tropical and Subtropical Botany,2014,22(4):383-390.
• [17]邓朝军,蒋际谋,龚慧文,许奇志,陈义勇,林永祥,郑少泉,邢建宏. 2个枇杷品种果皮日灼伤害差异蛋白质组分析[J].福建农业学报,2012,27(12):1313-1317.DENG Chaojun,JIANG Jimou,GONG Huiwen,XU Qizhi,CHEN Yiyong,LIN Yongxiang,ZHENG Shaoquan,XING Jianhong. Proteome analysis of loquat peel under sunburn in two varieties[J]. Fujian Journal of Agricultural Science,2012,27(12):1313-1317.
• [18]潘德灼,邓朝军,吕晓杰,宋晓敏,邱智敏,龚慧文,蒋际谋,陈伟.日灼胁迫对枇杷果皮蛋白质组的影响[J].西北植物学报,2016,36(9):1801-1812.PAN Dezhuo,DENG Chaojun,LüXiaojie,SONG Xiaomin,QIU Zhimin,GONG Huiwen,JIANG Jimou,CHEN Wei. Effect of sunburn stress on pericarp proteomics in loquat fruits[J]. Acta Botanica Boreali-Occidentalia Sinica,2016,36(9):1801-1812.
• [19]潘德灼,盖永红,吕晓杰,蒋雨露,邱智敏,宋晓敏,黄春梅,陈伟.龙眼果实采后果皮褐变过程差异表达蛋白分析[J].果树学报,2016,33(5):542-551.PAN Dezhuo,GE Yonghong,LüXiaojie,JIANG Yulu,QIU Zhimin,SONG Xiaomin,HUANG Chunmei,CHEN Wei. Analysis of differentially expressed proteins during the pericarp browning of postharvest longan fruit[J]. Journal of Fruit Science,2016,33(5):542-551.
• [20] BRADFORD M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry,1976,72:248-254.
• [21] YOU X,WANG L,LIANG W,GAI Y,WANG X,CHEN W.Floral reversion mechanism in longan(Dimocarpus longan Lour.)revealed by proteomic and anatomic analyses[J]. Journal of Proteomics,2012,75:1099-1118.
• [22]王学奎.植物生理生化实验原理与技术(第2版)[M].北京:高等教育出版社,2006:202-205,267-268.WANG Xuekui,The experiment principle and technique on plant physiology and biochemistry(Second Edition)[M]. Beijing:Higher Education Press,2006:202-205,267-268.
• [23]李利军,孔红星,伍时华,邹福声,陈林.蔗糖测定的紫外分光光度新方法研究及应用[J].分析科学学报,2003,19(4):367-369.LI Lijun,KONG Hongxin,WU Shihua,ZOU Fushen,CHENG Lin. A new UV-spectrophotometric method for the determination of sucrose[J]. Journal of Analytical Science,2003,19(4):367-369.
• [24]陈建勋,王晓峰.植物生理学实验指导(第二版)[M].广州:华南理工大学出版社,2006:81-84.CHEN Jianxun,WANG Xiaofeng. Experimental guidance for Plant Physiology(Second Edition)[M]. Guangzhou:South China University of Technology Press,2006:81-84.
• [25]高慧颖,王琦,陈源,谢鸿根,赖呈纯,余亚白.茂谷橘橙中类胡萝卜素含量的研究[J].福建农业学报,2010,25(2):197-200.GAO Huiying,WANG Qi,CHEN Yuan,XIE Honggen,LAI Chengchun,YU Yabai. Total carotenoids in murcott[J]. Fujian Journal of Agricultural Sciences,2010,25(2):197-200.
• [26] MORANTE-CARRIEL J,SELLéS-MARCHART S,MARTíNEZ-MáRQUEZ A,MARTíNEZ-ESTESO M J,LUQUE I,BRUMARTíNEZ R. RNA isolation from loquat and other recalcitrant woody plants with high quality and yield[J]. Analytical Biochemistry,2014,452:46-53.
• [27] JANECEK S,SVENSSON B,MACGREGOR E A. Alpha-amylase-an enzyme present in various sequence-based glycoside hydrolase families[J].FEBS Journal,2013,280:546.
• [28] WANG Y,ZHANG D. Activities,quantitative changes and subcellular localization ofα-amylase during development of apple fruit[J]. Acta Botanica sinica,,2002,44(1):34-41.
• [29] WEISE S E,SCHRADER S M,KLEINBECK K R,SHARKEY T D. Carbon balance and circadian regulation of hydrolytic and phosphorolytic breakdown of transitory starch[J]. Plant Physiology,2006,141(3):879-886.
• [30] MARTíNEZ-BARAJAS E,DELATTE T,SCHLUEPMANN H,DE JONG G J,SOMSEN G W,NUNES C,PRIMAVESI L F,COELLO P,MITCHELL R A,PAUL M J. Wheat grain development is characterized by remarkable trehalose 6-phosphate accumulation pregrain filling:tissue distribution and relationship to SNF1-related protein kinase1 activity[J]. Plant Physiology,2011,156(1):373-381.
• [31] BOWMAN E J,IKUMA H,STEIN H J. Citric acid cycle activity in mitochondria isolated from mung bean hypocotyls[J]. Plant Physiology,1976,58(3):426-432.
• [32] ARAúJO W L,NUNES-NESI A,TRENKAMP S,BUNIK V I,FERNIE A R. Inhibition of 2-oxoglutarate dehydrogenase in potato tuber suggests the enzyme is limiting for respiration and confirms its importance in nitrogen assimilation[J]. Plant Physiology,2008,148(4):1782-1796.
• [33] ARAúJO W L,TOHGE T,OSORIO S,LOHSE M,BALBO I,KRAHNERT I,SIENKIEWICZ-PORZUCEK A,USADEL B,NUNES-NESI A,FERNIE A R. Antisense inhibition of the 2-oxoglutarate dehydrogenase complex in tomato demonstrates its importance for plant respiration and during leaf senescence and fruit maturation[J]. The Plant Cell,2012,24(6):2328-2351.
• [34] ALBRECHT M,SANDMANN G. Light-stimulated carotenoid biosynthesis during transformation of maize etioplasts is regulated by increased activity of isopentenyl pyrophosphate isomerase[J]. Plant Physiology,1994,105(2):529-534.
• [35] GILIBERTO L,PERROTTA G,PALLARA P,WELLER J L,FRASER P D,BRAMLEY P M,FIORE A,TAVAZZA M,GIULIANO G.. Manipulation of the blue light photoreceptor cryptochrome 2 in tomato affects vegetative development,flowering time,and fruit antioxidant content[J]. Plant Physiology,2005,137(1):199-208.
• [36] KATO M,IKOMA Y,MATSUMOTO H,SUGIURA M,HYODO H,YANO M. Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit[J]. Plant Physiology,2004,134(2):824-837.
• [37] SHI H Y,ZHANG Y X. Pear ACO genes encoding putative 1-aminocyclopropane-1-carboxylate oxidase homologs are functionally expressed during fruit ripening and involved in response to salicylic acid[J]. Molecular Biology Reports,2012,39(10):9509-9519.