[1]羊杏平,张曼,徐锦华,等.西瓜幼苗根系发育过程中的蛋白质组分析[J].南京农业大学学报,2015,38(6):901-907.[doi:10.7685/j.issn.1000-2030.2015.06.005]
 YANG Xingping,ZHANG Man,XU Jinhua,et al.Proteomic analysis on root growth and development of watermelon seedlings[J].Journal of Nanjing Agricultural University,2015,38(6):901-907.[doi:10.7685/j.issn.1000-2030.2015.06.005]
点击复制

西瓜幼苗根系发育过程中的蛋白质组分析()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
38卷
期数:
2015年6期
页码:
901-907
栏目:
出版日期:
2015-11-18

文章信息/Info

Title:
Proteomic analysis on root growth and development of watermelon seedlings
作者:
羊杏平 张曼 徐锦华 刘广 姚协丰 李苹芳
江苏省农业科学院蔬菜研究所/江苏省高效园艺作物遗传改良重点实验室, 江苏 南京 210014
Author(s):
YANG Xingping ZHANG Man XU Jinhua LIU Guang YAO Xiefeng LI Pingfang
Institute of Vegetable Crops/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
关键词:
西瓜根系发育蛋白质组表达
Keywords:
watermelon(Citrullus lanatus)root developmentproteomicsgene expression
分类号:
S634.3
DOI:
10.7685/j.issn.1000-2030.2015.06.005
摘要:
[目的]通过对西瓜幼苗根系不同发育时期的蛋白质组变化的研究,从蛋白质表达水平上来揭示西瓜幼苗根系发育的分子机制.[方法]采用双向电泳技术对西瓜根系不同时期的蛋白质组进行比较,对差异蛋白质点进行图谱分析和MALDI-TOF/TOF质谱鉴定,采用荧光定量PCR技术(RT-qPCR)对候选蛋白基因进行表达验证.[结果]共得到12个差异表达蛋白质点,12个差异蛋白表现不同的表达趋势,其中上调表达4个,下调表达2个,不规则表达6个.经质谱分析,11个蛋白得到鉴定,对鉴定出的蛋白进行功能分析,其中10个蛋白分别涉及蛋白质生物合成(20%)、代谢相关(40%)、防御应答(30%)和GA信号途径(10%),1个为功能未知蛋白.4个候选蛋白基因的RT-qPCR分析结果显示,茉莉酸诱导蛋白1和抗坏血酸过氧化物酶基因在mRNA水平的表达变化与蛋白水平一致,而赤霉素受体基因GID1L2和茉莉酸诱导蛋白2基因的表达可能与转录后调控有关.[结论]本文揭示了西瓜幼苗根系发育蛋白质组的表达特征,获得了在西瓜根系发育过程中可能有重要功能的蛋白,为深入开展西瓜根系生长发育相关基因功能研究提供了数据资源.
Abstract:
[Objectives]To understand the function of differentially expressed proteins in roots of watermelon seedlings,proteins of watermelon roots at different developmental stages were studied using proteomics technique. This work will provide a theoretical basis for further studying on the molecular mechanism of watermelon root development. [Methods]A comparative study of watermelon roots at different developmental stages was carried out by using two-dimensional gel electrophoresis. The differentially expressed protein spots identified by electrophoresis were further analyzed by MALDI-TOF/TOF. Expression profiles of candidate protein genes were investigated using quantitative real time PCR(RT-qPCR). [Results]Twelve differentially expressed protein spots were identified by electrophoresis. Levels of 12 proteins were differential,of which,4 proteins were up-regulated,2 proteins were down-regulated and 6 proteins were differentially-regulated. 11 protein spots were finally identified by MALDI-TOF/TOF. Functions of these 11 proteins were analyzed which included 10 functional proteins involved in protein biosynthesis(20%),metabolism(40%),plant defense(30%)and GA signaling(10%),and 1 unknown protein. Four candidate protein genes were further investigated using RT-qPCR. Expression changes of jasmonate-induced protein 1 gene and ascorbate peroxidase gene in mRNA level were consistent with those in proteomic levels,while the expression of gibberellin receptor gene GID1L2 and Jasmonate-induced protein 2 gene might be related with transcriptional regulation. [Conclusions]This work gives a global insight into the characteristics of proteome of watermelon roots at different developmental stages by using two-dimensional gel electrophoresis. Proteins with important function were acquired which might be valuable resources for future research.

参考文献/References:

[1] Phizicky E,Bastiaens P I H,Zhu H,et al. Protein analysis on a proteomic scale[J]. Nature,2003,422(6928):208-215
[2] Shen S,Sharma A,Komatsu S. Characterization of proteins responsive to gibberellin in the leaf-sheath of rice(Oryza sativa L.)seedling using proteome analysis[J]. Biological and Pharmaceutical Bulletin,2003,26(2):129-136
[3] 范宝莉,王振英,陈宏,等. 小麦T型细胞质雄性不育系、保持系蛋白质双向电泳比较研究[J]. 实验生物学报,2004,37(1):45-49 [Fan B L,Wang Z Y,Chen H,et al. Comparative studies on proteins of cytoplasmic male-sterile wheat and its maintainer by 2D-PAGE in Triticum aestivum[J]. Acta Biologiae Experimentalis Sinica,2004,37(1):45-49(in Chinese with English abstract)]
[4] 李跃建,梁根云,刘小俊,等. 黄瓜嫁接苗和自根苗的蛋白质组学研究[J]. 园艺学报,2009,36(8):1147-1152 [Li Y J,Liang G Y,Liu X J,et al. Proteomic study on grafted and non-grafted cucumber(Cucumis sativus L.)[J]. Acta Horticulturae Sinica,2009,36(8):1147-1152(in Chinese with English abstract)]
[5] Schneider T,Gerrits B,Gassmann R,et al. Proteome analysis of fungal and bacterial involvement in leaf litter decomposition[J]. Proteomics,2010,10:1819-1830
[6] Malter D,Wolf S. Melon phloem-sap proteome:developmental control and response to viral infection[J]. Protoplasma,2011,248:217-224
[7] Guo S G,Zhang J G,Sun H H,et al. The draft genome of watermelon(Citrullus lanatus)and resequencing of 20 diverse accessions[J]. Nature Genetics,2013,45:51-58
[8] Yoshimura K,Masuda A,Kuwano M,et al. Programmed proteome response for drought avoidance/tolerance in the root of a C3 xerophyte(wild watermelon)under water deficits[J]. Plant and Cell Physiology,2008,49(2):226-241
[9] 杨冬冬. 西瓜砧木接穗互作的木质素合成和叶片蛋白质组分析[D]. 上海:上海交通大学,2006 [Yang D D. Lignin biosynthesis and leaf proteome analysis of watermelon rootstock-scion interactions[D]. Shanghai:Shanghai Jiao Tong University,2006(in Chinese with English abstract)]
[10] Yang Y J,Wang L P,Tian J,et al. Proteomic study participating the enhancement of growth and salt tolerance of bottle gourd rootstock-grafted watermelon seedlings[J]. Plant Physiology and Biochemistry,2012,58:54-65
[11] Jacobs D I,van Rijssen M S,van der Heijden R,et al. Sequential solubilization of proteins precipitated with trichloroacetic acid in acetone from cultured Catharanthus roseus cells yields 52% more spots after two-dimensional electrophoresis[J]. Proteomics,2001,1:1345-1350
[12] 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(1):248-254
[13] Arndt C,Koristka S,Feldmann A,et al. Coomassie-Brilliant blue staining of polyacrylamide gels[C]//Kurien B T,Hal Scofield R. Protein Electrophoresis:Methods and Protocols,Methods in Molecular Biology. Springer Science,2012,869:465-469
[14] 邵彩虹,王经源,林文雄. 苗期水稻叶片发育进程的差异蛋白质组学分析[J]. 中国农业科学,2008,41(11):3831-3837 [Shao C H,Wang J Y,Lin W X. Differential proteomics analysis of leaf development at rice(Oryza sativa)seedling stage[J]. Scientia Agricultura Sinica,2008,41(11):3831-3837(in Chinese with English abstract)]
[15] Clemens M J. Targets and mechanisms for the regulation of translation in malignant transformation[J]. Oncogene,2004,23:3180-3188
[16] 栗现芳,马守才,张改生,等. 小麦核糖体蛋白S15a基因(TaRPS15a)的克隆及其在多子房株系中的时空表达分析[J]. 农业生物技术学报,2011,19(2):236-242 [Li X F,Ma S C,Zhang G S,et al. Cloning of ribosomal protein S15a gene(TaRPS15a)and its expression patterns based on temporal-spatial in multi-ovary line of wheat(Triticum aestivum)[J]. Journal of Agricultural Biotechnology,2011,19(2):236-242(in Chinese with English abstract)]
[17] 杜军志,张会梅,杨绒会. 环境条件对西瓜生长发育的影响[J]. 中国西瓜甜瓜,2004(5):15-17 [Du J Z,Zhang H M,Yang R H. Effect of environmental condition on watermelon growth and development[J]. China Watermelon and Melon,2004(5):15-17(in Chinese)]
[18] Cheong J J,Choi Y D. Methyl jasmonate as a vital substance in plants[J]. Trends in Genetics,2003,19(7):409-413
[19] Bona E,Marsano F,Cavaletto M,et al. Proteomic characterization of copper stress response in Cannabis sativa roots[J]. Proteomics,2007,7:1121-1130
[20] Turóczy Z,Kis P,Török K,et al. Overproduction of a rice aldo-keto reductase increases oxidative and heat stress tolerance by malondialdehyde and methylglyoxal detoxification[J]. Plant Molecular Biology,2011,75(4/5):399-412
[21] 郑建树,喻春明,陈平,等. 苎麻谷氨酰胺合成酶BnGS2等位基因的克隆及其转基因烟草特性[J]. 中国农业科学,2014,47(17):3348-3358 [Zheng J S,Yu C M,Chen P,et al. Cloning of glutamine synthetase BnGS2 allele genes from ramie(Boehmeria nivea L.)and study on gene-transforming tobacco[J]. Scientia Agricultura Sinica,2014,47(17):3348-3358(in Chinese with English abstract)]
[22] Jan A,Nakamura H,Handa H,et al. Gibberellin regulates mitochondrial pyruvate dehydrogenase activity in rice[J]. Plant and Cell Physiology,2006,47(2):244-253
[23] 王丽,王芹芹,王幼群. 蚕豆叶片小叶脉不同发育时期ATP酶和酸性磷酸酶的细胞化学超微结构定位[J]. 植物学报,2014,49(1):78-86 [Wang L,Wang Q Q,Wang Y Q. Cytochemical localization of ATPase and acid phosphatase in minor veins of the leaf of Vicia faba during different developmental stages[J]. Chinese Bulletin of Botany,2014,49(1):78-86(in Chinese with English abstract)]
[24] Hu L X,Li H Y,Pang H C,et al. Responses of antioxidant gene,protein and enzymes to salinity stress in two genotypes of perennial ryegrass(Lolium perenne)differing in salt tolerance[J]. Journal of Plant Physiology,2012,169:146-156
[25] 李强,吴建明,梁和,等. 高等植物赤霉素生物合成及其信号转导途径[J]. 生物技术通报,2014(10):16-22 [Li Q,Wu J M,Liang H,et al. Gibberellins biosynthesis and signaling transduction pathway in higher plant[J]. Biotechnology Bulletin,2014(10):16-22(in Chinese with English abstract)]
[26] Baldi P,Long A D. A Bayesian framework for the analysis of microarray expression data:regularized t-test and statistical inferences of gene changes[J]. Bioinformatics,2001,17:509-519

相似文献/References:

[1]郝文雅,沈其荣,冉炜,等.西瓜和水稻根系分泌物中糖和氨基酸对西瓜枯萎病病原菌生长的影响[J].南京农业大学学报,2011,34(3):77.[doi:10.7685/j.issn.1000-2030.2011.03.014]
 HAO Wen-ya,SHEN Qi-rong,RAN Wei,et al.The effects of sugars and amino acids in watermelon and rice root exudates on the growth of Fusarium oxysporum f.sp. niveum[J].Journal of Nanjing Agricultural University,2011,34(6):77.[doi:10.7685/j.issn.1000-2030.2011.03.014]
[2]苏世鸣,任丽轩,杨兴明,等.西瓜专化型尖孢镰刀菌的分离鉴定及水稻根系分泌物对其生长的影响[J].南京农业大学学报,2008,31(1):57.[doi:10.7685/j.issn.1000-2030.2008.01.012]
 SU Shi-ming,REN Li-xuan,YANG Xing-ming,et al.Identification of the Fusarium oxysporum f.sp. niveum and the effects of rice root exudates on its growth[J].Journal of Nanjing Agricultural University,2008,31(6):57.[doi:10.7685/j.issn.1000-2030.2008.01.012]
[3]韩志平,郭世荣,冯吉庆,等.盐胁迫对西瓜幼苗生长、叶片光合色素和脯氨酸含量的影响[J].南京农业大学学报,2008,31(2):32.[doi:10.7685/j.issn.1000-2030.2008.02.007]
 HAN Zhi-ping,GUO Shi-rong,FENG Ji-qing,et al.Effect of salinity on plant growth,photosynthetic pigments and proline content in leaves of watermelon seedlings[J].Journal of Nanjing Agricultural University,2008,31(6):32.[doi:10.7685/j.issn.1000-2030.2008.02.007]
[4]吕卫光,黄启为,沈其荣,等.不同来源有机肥及有机肥与无机肥混施对西瓜生长期土壤酶活性的影响[J].南京农业大学学报,2005,28(4):68.[doi:10.7685/j.issn.1000-2030.2005.04.015]
[5]薛宝娣,陈永萱,刘凤权,等.江苏省西瓜花叶病毒类型的鉴定[J].南京农业大学学报,1987,10(04):53.[doi:10.7685/j.issn.1000-2030.1987.04.009]
 Xue Baodi,Chen Yongxuan,Liu Fengquan,et al.IDENTIFICATION OF WATERMELON MOSAIC VIRUSES IN JIANGSU PROVINCE[J].Journal of Nanjing Agricultural University,1987,10(6):53.[doi:10.7685/j.issn.1000-2030.1987.04.009]
[6]张振兴,孙锦,郭世荣*,等.增施钙素对盐胁迫下西瓜幼苗生长和可溶性蛋白含量及组分的影响[J].南京农业大学学报,2011,34(5):20.[doi:10.7685/j.issn.1000-2030.2011.05.004]
 ZHANG Zhen-xing,SUN Jin,GUO Shi-rong*,et al.Effect of supplementary calcium on the growth and the content and components of soluble protein of watermelon seedlings under salt-stress[J].Journal of Nanjing Agricultural University,2011,34(6):20.[doi:10.7685/j.issn.1000-2030.2011.05.004]
[7]赵鹏,董彩霞,申长卫,等.3种有机无机肥配施对西瓜氮、钾养分吸收以及产量和品质的影响[J].南京农业大学学报,2015,38(2):288.[doi:10.7685/j.issn.1000-2030.2015.02.017]
 ZHAO Peng,DONG Caixia,SHEN Changwei,et al.Effects of three organic and inorganic fertilizers combined application on absorption of nitrogen and potassium,yield and quality of watermelon[J].Journal of Nanjing Agricultural University,2015,38(6):288.[doi:10.7685/j.issn.1000-2030.2015.02.017]
[8]沈怡斐,鄂垚瑶,阳芳,等.西瓜根系分泌物中氨基酸组分对多黏类芽孢杆菌SQR-21趋化性及根际定殖的影响[J].南京农业大学学报,2017,40(1):101.[doi:10.7685/jnau.201603051]
 SHEN Yifei,E Yaoyao,YANG Fang,et al.Effects of amino acids in root exudates of watermelon on the chemotactic reaction and root colonization of Paenibacillus polymyxa SQR-21[J].Journal of Nanjing Agricultural University,2017,40(6):101.[doi:10.7685/jnau.201603051]
[9]浩折霞,黄大鹏,顾少华,等.酒糟-牛粪堆肥复配瓜果类蔬菜育苗基质配方筛选[J].南京农业大学学报,2017,40(3):457.[doi:10.7685/jnau.201605024]
 HAO Zhexia,HUANG Dapeng,GU Shaohua,et al.Screening of melon and fruit of vegetables grown substrate from different vinasse and cow dung composts mixtures[J].Journal of Nanjing Agricultural University,2017,40(6):457.[doi:10.7685/jnau.201605024]
[10]董亚,王硕,吴萍,等.西瓜钾素吸收特征及钾肥施用优化机制[J].南京农业大学学报,2018,41(1):98.[doi:10.7685/jnau.201702028]
 DONG Ya,WANG Shuo,WU Ping,et al.Absorption character of potassium and optimization mechanism of potassic fertilization on watermelon[J].Journal of Nanjing Agricultural University,2018,41(6):98.[doi:10.7685/jnau.201702028]

备注/Memo

备注/Memo:
收稿日期:2015-6-5。
基金项目:国家西甜瓜产业技术体系项目(CARS-No.8);江苏省重大科技支撑计划项目(BE2012323);江苏省农业科技自主创新资金项目[CX(14)2004]
作者简介:羊杏平,研究员,硕导,主要从事西瓜、甜瓜育种研究,E-mail:xingping@jaas.ac.cn.
更新日期/Last Update: 1900-01-01