枣果实不同发育阶段蛋白质组动态研究
|
摘要
枣果实在发育和成熟过程中涉及多种复杂的生理生化变化。为了解枣果实在发育过程中蛋白质组的变化情况,利用液质联用(LC-MS)技术对主栽制干枣品种——骏枣幼果期、膨大期、白熟期、初红期和全红期5个阶段的果实进行了非标记定量蛋白质组动态变化研究。结果表明,各时期果实与其前一阶段相比分别有差异表达蛋白189、356、82、69个。基因本体(GO)注释结果显示:幼果与膨大果的差异蛋白主要富集在核酸代谢过程,膨大期果实与白熟果的差异蛋白主要富集在碳水化合物代谢过程。从5个阶段的果实中分别鉴定到604、186、91、83、189个特异表达蛋白。Mercator注释和KEGG通路注释结果显示,幼果期特异表达蛋白主要参与了RNA转运通路。加权基因共表达网络(WGCNA)分析显示,幼果期高表达蛋白主要与核糖体合成相关。总之,枣果实在白熟期以前与细胞分裂和膨大相关的蛋白表达量和富集程度高,从白熟期开始与碳水化合物积累相关的蛋白质表达上调。本研究从蛋白质组学水平阐明了枣果实发育成熟过程中蛋白质组分的动态变化,为提高枣果实品质奠定了理论基础。
In order to understand the dynamic process of the proteomics during the progress of jujube fruit development and ripening,we quantitatively analyzed the proteomics of jujube fruits at 5 developmental stages(young,enlarging,white mature,early red,and fully red) using high performance liquid chromatography-mass spectrometry method.The results showed that compared with the former stage,there existed 189,356,82,69 differentially expressed proteins,respectively in the fruit of the last four stages.The gene ontology annotation showed that the differential proteins between young and enlarging fruit mainly enriched in nucleic acid metabolism,and the differential proteins between enlarging fruit and white mature fruit were mainly involved in carbohydrate metabolism.We also identified 604,186,91,83,189 expressed proteins from the fruit of five stages,respectively.Mercator annotation and KEGG pathway annotation revealed that those specifically expressed proteins in young fruit were mainly involved in the RNA transport pathway.WGCNA analysis showed that the highly expressed proteins in the young fruit were mainly related to ribosome biogenesis.In general,prior to the white mature stage,the jujube fruit mainly underwent cell division and enlargement,while the fruit mainly underwent carbohydrate accumulation after the white ripening stage.The dynamic of proteins during jujube fruit development and maturity were elucidated from proteomics level,and therefore laid a theoretical foundation for improving the quality of jujube fruit.
In order to understand the dynamic process of the proteomics during the progress of jujube fruit development and ripening,we quantitatively analyzed the proteomics of jujube fruits at 5 developmental stages(young,enlarging,white mature,early red,and fully red) using high performance liquid chromatography-mass spectrometry method.The results showed that compared with the former stage,there existed 189,356,82,69 differentially expressed proteins,respectively in the fruit of the last four stages.The gene ontology annotation showed that the differential proteins between young and enlarging fruit mainly enriched in nucleic acid metabolism,and the differential proteins between enlarging fruit and white mature fruit were mainly involved in carbohydrate metabolism.We also identified 604,186,91,83,189 expressed proteins from the fruit of five stages,respectively.Mercator annotation and KEGG pathway annotation revealed that those specifically expressed proteins in young fruit were mainly involved in the RNA transport pathway.WGCNA analysis showed that the highly expressed proteins in the young fruit were mainly related to ribosome biogenesis.In general,prior to the white mature stage,the jujube fruit mainly underwent cell division and enlargement,while the fruit mainly underwent carbohydrate accumulation after the white ripening stage.The dynamic of proteins during jujube fruit development and maturity were elucidated from proteomics level,and therefore laid a theoretical foundation for improving the quality of jujube fruit.
引文
[1] 中国林业统计年鉴委员会.中国林业统计年鉴(2016)[M].北京:中国林业出版社,2017:86-87.
[2] 樊保国,李月梅,李登科.鲜食枣品质性状的综合评价[J].西北林学院学报,2012,27(2):79-82,87.FAN B G,LI Y M,LI D K.Comprehensive assessmet of the quality characters of fresh-jujube cultivars[J].Journal of Northwest Forestry University,2012,27(2):79-82,87.(in Chinese)
[3] 李欢,张舒怡,张钟,等.鲜食枣与制干枣的成熟软化机理差异研究[J].西北林学院学报,2017,32(5):137-143.LI H,ZHANG S Y,ZHANG Z,et al.Differences in ripening and softening mechanism between fresh-eating and dried jujube fruit[J].Journal of Northwest Forestry University,2017,32(5):137-143.(in Chinese)
[4] 曲泽洲,王永惠,周吉柱,等.枣果实发育与细胞分裂和增长的关系[J].河北农业大学学报,1964,3(1):1-15.
[5] 张怀龙,马峰秀.枣果实发育动态规律与优化栽培的相关性[J].林业科技通讯,1997(3):22-23.
[6] 费光雪.骏枣果实糖分积累及相关代谢酶活性变化的研究[J].新疆农垦科技,2018,41(4):36-39.
[7] 林思思,蒲小秋,费光雪,等.冬枣和骏枣果实发育过程中糖组分及代谢酶活性变化的研究[J].黑龙江农业科学,2018(7):8-11.
[8] SHI Q Q,ZHANG Z,SU J J,et al.Comparative analysis of pigments,phenolics,and antioxidant activity of Chinese jujube (Ziziphus jujuba Mill.) during fruit development[J].Molecules,2018,23(8):1917.
[9] 张琼,周广芳,沈广宁,等.冬枣果皮着色过程中类黄酮类物质成分及含量的变化[J].园艺学报,2010,37(2):193-198.ZHANG Q,ZHOU G F,SHEN G N,et al.The flavonoids in the fruit peel of Ziziphus jujuba Mill.‘Dongzao’ during coloring process [J].Acta Horticulturae Sinica,2010,37(2):193-198.(in Chinese)
[10] 张春梅.枣糖酸代谢及其驯化的分子机制研究[D].杨陵:西北农林科技大学,2016.
[11] 魏琦琦.基于转录组测序的枣成花和果实品质重要基因的研究[D].长沙:中南林业科技大学,2017.
[12] 解振强,韩柏明,巫建华.葡萄着色期果皮蛋白质组分析[J].园艺学报,2018,45(11):1-22.XIE Z Q,HAN B M,WU J H.Proteome analysis of during grape coloring period[J].Acta Horticulturae Sinica,2018,45(11):1-22.(in Chinese)
[13] 董飞,王传增,孙秀东,等.基于蛋白质组学研究红光对番茄果实糖代谢的影响[J].园艺学报,2018,45(10):78-88.DONG F,WANG C Z,SUN X D,et al.Studies on the sugar metabolism mechanism of tomato fruit under red light revealed by proteomic analysis[J].Acta Horticulturae Sinica,2018,45(10):78-88.(in Chinese)
[14] 周凯凯,张胜,赵忠.不同树龄银杏叶片差异蛋白组学研究[J].西北林学院学报,2018,33(1):105-112.ZHOU K K,ZHANG S,ZHAO Z.Proteomic analysis on ginkgo leaves with different ages[J].Journal of Northwest Forestry University,2018,33(1):105-112.(in Chinese)
[15] PAN X Q,ZHU B Z,ZHU H L,et al.iTRAQ protein profile analysis of tomato green-ripe mutant reveals new aspects critical for fruit ripening[J].Journal of Proteome Research,2014,13(4):1979-1993.
[16] 孙倩倩.外源褪黑素对番茄果实采后成熟的影响[D].北京:中国农业大学,2016.
[17] CHEN R H,CHEN G L,HUANG J.Shot-gun proteome and transcriptome mapping of the jujube floral organ and identification of a pollen-specific S-locus F-box gene[J].Peerj,2017,5:e3588.
[18] HUANG J,ZHANG C,ZHAO X,et al.The jujube genome provides insights into genome evolution and the domestication of sweetness/acidity taste in fruit trees[J].PLOS Genetics,2016,12(12):e1006433.
[19] STEFAN G,JUAN M G,TEROL J,et al.High-throughput functional annotation and data mining with the Blast2GO suite[J].Nucleic Acids Research,2008,36(10):3420-3435.
[20] 曹尚银,沈程清,曹达,等.盐胁迫下枣叶片蛋白质组差异的分析[J].果树学报,2013,30(1):43-47,185.
[21] 金大伟,杨军,李锋,等.烟草非特异性脂质转移蛋白基因的克隆与表达分析[J].烟草科技,2015,48(1):12-20.
[22] 桑利丹.甘油醛三磷酸脱氢酶参与蜡样芽孢杆菌0-9生物膜形成[D].开封:河南大学,2018.
[23] 卜娇迪,陈莹莹,刘孟军,等.枣甘油醛-3-磷酸脱氢酶基因的克隆及表达分析[J].北方园艺,2015(13):102-106.BU J D,CHEN Y Y,LIU M J,et al.Cloning and expression analysis of glyceraldehyde-3-phosphate dehydrogenase from Ziziphus jujuba[J].Northern Horticulture,2015(13):102-106.(in Chinese)
[2] 樊保国,李月梅,李登科.鲜食枣品质性状的综合评价[J].西北林学院学报,2012,27(2):79-82,87.FAN B G,LI Y M,LI D K.Comprehensive assessmet of the quality characters of fresh-jujube cultivars[J].Journal of Northwest Forestry University,2012,27(2):79-82,87.(in Chinese)
[3] 李欢,张舒怡,张钟,等.鲜食枣与制干枣的成熟软化机理差异研究[J].西北林学院学报,2017,32(5):137-143.LI H,ZHANG S Y,ZHANG Z,et al.Differences in ripening and softening mechanism between fresh-eating and dried jujube fruit[J].Journal of Northwest Forestry University,2017,32(5):137-143.(in Chinese)
[4] 曲泽洲,王永惠,周吉柱,等.枣果实发育与细胞分裂和增长的关系[J].河北农业大学学报,1964,3(1):1-15.
[5] 张怀龙,马峰秀.枣果实发育动态规律与优化栽培的相关性[J].林业科技通讯,1997(3):22-23.
[6] 费光雪.骏枣果实糖分积累及相关代谢酶活性变化的研究[J].新疆农垦科技,2018,41(4):36-39.
[7] 林思思,蒲小秋,费光雪,等.冬枣和骏枣果实发育过程中糖组分及代谢酶活性变化的研究[J].黑龙江农业科学,2018(7):8-11.
[8] SHI Q Q,ZHANG Z,SU J J,et al.Comparative analysis of pigments,phenolics,and antioxidant activity of Chinese jujube (Ziziphus jujuba Mill.) during fruit development[J].Molecules,2018,23(8):1917.
[9] 张琼,周广芳,沈广宁,等.冬枣果皮着色过程中类黄酮类物质成分及含量的变化[J].园艺学报,2010,37(2):193-198.ZHANG Q,ZHOU G F,SHEN G N,et al.The flavonoids in the fruit peel of Ziziphus jujuba Mill.‘Dongzao’ during coloring process [J].Acta Horticulturae Sinica,2010,37(2):193-198.(in Chinese)
[10] 张春梅.枣糖酸代谢及其驯化的分子机制研究[D].杨陵:西北农林科技大学,2016.
[11] 魏琦琦.基于转录组测序的枣成花和果实品质重要基因的研究[D].长沙:中南林业科技大学,2017.
[12] 解振强,韩柏明,巫建华.葡萄着色期果皮蛋白质组分析[J].园艺学报,2018,45(11):1-22.XIE Z Q,HAN B M,WU J H.Proteome analysis of during grape coloring period[J].Acta Horticulturae Sinica,2018,45(11):1-22.(in Chinese)
[13] 董飞,王传增,孙秀东,等.基于蛋白质组学研究红光对番茄果实糖代谢的影响[J].园艺学报,2018,45(10):78-88.DONG F,WANG C Z,SUN X D,et al.Studies on the sugar metabolism mechanism of tomato fruit under red light revealed by proteomic analysis[J].Acta Horticulturae Sinica,2018,45(10):78-88.(in Chinese)
[14] 周凯凯,张胜,赵忠.不同树龄银杏叶片差异蛋白组学研究[J].西北林学院学报,2018,33(1):105-112.ZHOU K K,ZHANG S,ZHAO Z.Proteomic analysis on ginkgo leaves with different ages[J].Journal of Northwest Forestry University,2018,33(1):105-112.(in Chinese)
[15] PAN X Q,ZHU B Z,ZHU H L,et al.iTRAQ protein profile analysis of tomato green-ripe mutant reveals new aspects critical for fruit ripening[J].Journal of Proteome Research,2014,13(4):1979-1993.
[16] 孙倩倩.外源褪黑素对番茄果实采后成熟的影响[D].北京:中国农业大学,2016.
[17] CHEN R H,CHEN G L,HUANG J.Shot-gun proteome and transcriptome mapping of the jujube floral organ and identification of a pollen-specific S-locus F-box gene[J].Peerj,2017,5:e3588.
[18] HUANG J,ZHANG C,ZHAO X,et al.The jujube genome provides insights into genome evolution and the domestication of sweetness/acidity taste in fruit trees[J].PLOS Genetics,2016,12(12):e1006433.
[19] STEFAN G,JUAN M G,TEROL J,et al.High-throughput functional annotation and data mining with the Blast2GO suite[J].Nucleic Acids Research,2008,36(10):3420-3435.
[20] 曹尚银,沈程清,曹达,等.盐胁迫下枣叶片蛋白质组差异的分析[J].果树学报,2013,30(1):43-47,185.
[21] 金大伟,杨军,李锋,等.烟草非特异性脂质转移蛋白基因的克隆与表达分析[J].烟草科技,2015,48(1):12-20.
[22] 桑利丹.甘油醛三磷酸脱氢酶参与蜡样芽孢杆菌0-9生物膜形成[D].开封:河南大学,2018.
[23] 卜娇迪,陈莹莹,刘孟军,等.枣甘油醛-3-磷酸脱氢酶基因的克隆及表达分析[J].北方园艺,2015(13):102-106.BU J D,CHEN Y Y,LIU M J,et al.Cloning and expression analysis of glyceraldehyde-3-phosphate dehydrogenase from Ziziphus jujuba[J].Northern Horticulture,2015(13):102-106.(in Chinese)