魏蓉;巩培杰;李树秀;王跃进;张朝红;

• 1:西北农林科技大学园艺学院·旱区作物逆境生物学国家重点实验室·农业部西北地区园艺作物生物学与种质创制重点实验室

摘要(Abstract)：

【目的】研究葡萄类TIAR蛋白与无核葡萄胚珠败育的关系,探索无核葡萄胚珠败育分子机理。【方法】以‘无核白’葡萄(Vitis vinifera‘Thompson Seedless’)浆果发育过程中差异表达的EST序列为基础,克隆目的基因全长序列,并通过实时荧光定量PCR技术分析了该基因在无核葡萄品种‘无核白’和有核葡萄品种‘黑比诺’胚珠发育过程中的表达变化。【结果】获得一条1 593 bp的同源序列,命名为VvTIAR-like(GenBank登录号:KF523863),其ORF长1 269 bp,编码422个氨基酸,蛋白理论分子质量为45.8 kD。生物信息学分析显示所得基因具有典型的TIAR蛋白质特征,与其他植物来源的类TIAR蛋白也具有较高同源性,认为该序列为编码葡萄类TIAR蛋白的基因。实时荧光定量PCR技术分析结果显示,VvTIAR-like在‘无核白’与‘黑比诺’胚珠发育过程中表现出不同的表达模式。在‘无核白’胚珠开始败育的前期和关键时期,该基因表现出明显的表达高峰,而在‘黑比诺’中则表达量低且无明显变化。【结论】获得了‘无核白’葡萄中类TIAR基因VvTIAR-like的cDNA全长,对该基因在无核葡萄品种‘无核白’和有核葡萄品种‘黑比诺’胚珠发育过程中的表达分析显示,VvTIAR-like可能与‘无核白’葡萄胚珠败育相关。

关键词(KeyWords)： 葡萄;类TIAR;基因克隆;表达分析

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31171926);; 国家现代农业产业技术体系建设专项(CARS-30-yz-7);; 陕西省农业攻关项目(2013K01-28)

作者(Author): 魏蓉;巩培杰;李树秀;王跃进;张朝红;

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参考文献(References)：

• [1]JIANG Jian-fu,SUN Hai-sheng,LIU Chong-huai,FAN Xiu-cai,ZHANG Ying.Progress in the research of grape breeding from 2000in China[J].Sino-Overseas Grapevine and Wine,2010(3):60-65,69.姜建福,孙海生,刘崇怀,樊秀彩,张颖.2000年以来中国葡萄育种研究进展[J].中外葡萄与葡萄酒,2010(3):60-65,69.
• [2]KAWAKAMI A,TIAN Q,DUAN X,STREULI M,SCHLOSSMAN S F,ANDERSON P.Identification and functional characterization of a TIA-1-related nucleolysin[J].PNAS,1992,89(18):8681-8685.
• [3]ISENI F,GARCIN D,NISHIO M,KOLAKOFSKY D.Sendai virus trailer RNA binds TIAR,a cellular protein involved in virus-induced apoptosis[J].EMBO J,2002,21(19):5141-5150.
• [4]TAUPIN J,TIAN Q,KEDERSHA N,ROBERTSOn M,ANDERSON P.The RNA-binding protein TIAR is translocated from the nucleus to the cytoplasm during Fas-mediated apoptotic cell death[J].PNAS,1995,92(5):1629-1633.
• [5]FRCH P,PUIG O,KEDERSHa N,MARTINEZ C,GRANNEMAN S,SRAPHIN B,ANDERSON P,VALCRCEL J.The apop-tosis-promoting factor TIA-1 is a regulator of alternative pre-mRNA splicing[J].Mol Cell,2000,6(5):1089-1098.
• [6]DEL-GATTO F,PLET A,GESNEL M C,FORT C,BREATHNACH R.Multiple interdependent sequence elements control splicing of a fibroblast growth factor receptor 2 alternative exon[J].Mol Cell Biol,1997,17(9):5106-5116.
• [7]ZHU H,HASMAN R A,YOUNG K M,KEDERSHA N L,LOU H.U1 snRNP-dependent function of TIAR in the regulation of alternative RNA processing of the human calcitonin/CGRP pre-mRNA[J].Mol Cell Biol,2003,23(17):5959-5971.
• [8]YU Q,COK S J,ZENG C,MORRISON A R.Translational repression of human matrix metalloproteinases-13 by an alternatively spliced form of T-cell-restricted intracellular antigen-related protein(TIAR)[J].J Biol Chem,2003,278(3):1579-1584.
• [9]KANDASAMY K,JOSEPH K,SUBRAMANIAM K,RAYMOND J R,THOLANIKUNNEL B G.Translational control ofβ2-adrenergic receptor mRNA by T-cell-restricted intracellular antigen-related protein[J].J Biol Chem,2005,280(3):1931-1943.
• [10]LIAO B,HU Y,BREWER G.Competitive binding of AUF1 and TIAR to MYC mRNA controls its translation[J].Nat Struct Mol Biol,2007,14(6):511-518.
• [11]KEDERSHA N,ANDERSON P.Stress granules:sites of mRNA triage that regulate mRNA stability and translatability[J].Biochem Soc Trans,2002,30(6):963-969.
• [12]BECK A R,MILLER J J,ANDERSON P,STREULI M.RNAbinding protein TIAR is essential for primordial germ cell development[J].PNAS,1998,95(5):2331-2336.
• [13]SNCHEZ-JIMNEZ C,IZQUIERDO J M.T-cell Intracellular Antigen(TIA)-Proteins Deficiency in Murine Embryonic Fibroblasts Alters Cell Cycle Progression and Induces Autophagy[J/OL].PloS one,2013,[2013-10-31].http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0075127#pone-0075127-g008.
• [14]KHARRAZ Y,SALMAND P A,CAMUS A,AURIOL J,GUEYDAN C,KRUYS V,MORELLO D.Impaired embryonic development in mice overexpressing the RNA-binding protein TIAR[J/OL].PLoS One,2010,[2013-10-31].http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0011352.
• [15]COSTENARO-DA-SILVA D,PASSAIA G,HENRIQUES J A.Identification and expression analysis of genes associated with the early berry development in the seedless grapevine(Vitis vinifera L.)cultivar Sultanine[J].Plant Sci,2010,179(5):510-519.
• [16]ZHANG Jin-jin,WANG Yue-jin,WANG Xi-ping,YANG Keqiang,YANG Jin-xiao.An improved method for rapidly extracting total RNA from vitis[J].Journal of Fruit Science,2003,20(3):178-181.张今今,王跃进,王西平,杨克强,杨进孝.葡萄总RNA提取方法的研究[J].果树学报,2003,20(3):178-181.
• [17]LI Shu-xiu,ZHANG Chao-hong,GONG Pei-jie,YANG Kun,ZHANG Xu-tong,WANG Yue-jin.Cloning and expression analysis of the differentially expressed VvWRKY20 Gene in the development of seedlessness grape(Vitis vinifera)ovule abortion[J].Journal of Agricultural Biotechnology,2012,20(10):1143-1149.李树秀,张朝红,巩培杰,杨堃,张旭彤,王跃进.无核葡萄VvWRKY20基因的克隆及其在珠败育过程中的差异表达分析[J].农业生物技术学报,2012,20(10):1143-1149.
• [18]DEO A,CARLSSON J,LINDLF A.How to choose a normalization strategy for miRNA quantitative real-time(qPCR)arrays[J].Journal of Bioinformatics and Computational Biology,2011,9(6):795-812.
• [19]SUN Bo,CAO Hou-nan,HE Wei,GONG Yan-ming,ZONG Changling.Study on embryo abortion stage of pseudoparthenocarpy seedless grape[J].Journal of Agricultural Science Yanbian University,2009,3(2):159-164.孙博,曹后男,何伟,巩艳明,宗长玲.假单性结实无核葡萄胚败育时期的研究[J].延边大学农学学报,2009,3(2):159-164.