山葡萄COR413家族基因克隆及其参与低温胁迫的表达分析

发布时间：2018-05-07 08:32

  本文选题：抗寒性 + 冷适应蛋白基因(COR)　； 参考：《农业生物技术学报》2017年03期

【摘要】：冷适应蛋白(cold-regulated,COR413)基因作为植物特有的一类低温胁迫响应功能基因,在抗寒过程中发挥重要作用。为了鉴定葡萄属(Vitis)植物COR413基因参与低温胁迫响应的功能特点,本研究基于欧洲葡萄基因组数据库挖掘,采用同源克隆法获得了抗寒种质山葡萄(Vitis amurensis)COR413基因家族成员的4条序列,分别命名为Va COR413-PM2A、-PM1X1、-IM1与-PM2B。经序列分析表明,所克隆的4条山葡萄COR413基因家族成员与欧洲葡萄相应基因的核苷酸与氨基酸序列高度同源,其核苷酸序列的差异仅为个别单核苷酸的替代。所克隆的4个山葡萄冷适应蛋白家族基因均含有WCOR413保守结构域、跨膜结构域与丝/苏/酪氨酸磷酸化位点,其中仅Va COR413-PM1X1含有一个预测的N端信号肽序列,仅Va COR413-PM2B包含1个糖基化磷脂酰肌醇锚点。源于2个葡萄种质中COR413基因家族成员可以聚类为2个明显的簇,COR413-PM2A,-PM1X1与-PM2B聚类在一簇,而COR413-IM1分布在另外一簇。半定量RT-PCR组织特异性检测结果表明,Va COR413-PM2A、-PM1X1、-IM1与-PM2B在山葡萄不同组织和器官中表达存在差异,且不同于欧洲葡萄相应基因的表达模式;其中山葡萄Va COR413-PM2A、-PM1X1、-IM1与-PM2B在所有检测的组织中均表达且强度不同,Va COR413-PM2A与-PM1X1在嫩梢、卷须、花絮以及幼叶中表达量较高。在低温胁迫条件下,山葡萄COR413基因家族4个成员参与低温胁迫的表达模式分为2类,而欧洲葡萄中相应成员有3种模式;山葡萄Va COR413-IM1受低温胁迫24 h表达量最高,为对照的9.57倍,而欧洲葡萄Vv COR413-IM1在低温胁迫3 h后的表达量较高,约为对照的3.65倍。低温胁迫下的表达数据表明,山葡萄COR413基因家族成员中聚类为一簇的Va COR413-PM2A,-PM1X1与-PM2B响应低温总体呈下调表达,其表达幅度均低于欧洲葡萄相应基因的表达量,而聚类在另外一簇的Va COR413-IM1对低温胁迫(24 h)响应诱导表达显著;研究结果为探明该基因家族成员在葡萄响应低温胁迫过程中的可能功能和作用机制提供了参考依据,同时为葡萄属植物抗寒(冻)性分子改良提供了理论依据。
[Abstract]:Cold-regulated Cor413) gene plays an important role in the process of cold resistance as a kind of plant specific low temperature stress response gene. In order to identify the functional characteristics of COR413 gene in Vitis plants involved in response to low temperature stress, four sequences of amurensis)COR413 gene family members were obtained by homologous cloning based on the European grape genome database. They were named Va Corr 413-PM2AU -PM1X1C1-IM1 and -PM2B respectively. The sequence analysis showed that the cloned COR413 gene family members were highly homologous to the nucleotide and amino acid sequences of the corresponding genes of European grape, and the difference of nucleotide sequence was only the substitution of individual single nucleotides. The four genes of cold acclimated protein family of Vitis vinifera contained WCOR413 conserved domain, transmembrane domain and silk / S / tyrosine phosphorylation site. Only Va COR413-PM1X1 contained a predicted N-terminal signal peptide sequence. Only Va COR413-PM2B contains one glycosylphosphatidylinositol anchor. From two grape germplasms, the COR413 gene family members can be clustered into two distinct clusters: Cor413-PM2An-PM1X1 and -PM2B in one cluster, and COR413-IM1 in another cluster. The results of semi-quantitative RT-PCR tissue specific detection showed that Va Cor413-PM2AM-PM1X1-IM1 and -PM2B were different in different tissues and organs, and different from the corresponding gene expression patterns of European grape. Va Cor413-PM2An-PM1X1C1-IM1 and -PM2B were expressed in all tissues examined, and the expression of Va COR413-PM2A and -PM1X1 were higher in young shoots, tendrils, floss and young leaves. Under the condition of low temperature stress, four members of COR413 gene family of Vitis vinifera were divided into two groups of expression patterns, while the corresponding members of European grape had three patterns, and Va COR413-IM1 was the most expressed in 24 h under low temperature stress. The expression of VV COR413-IM1 in European grape was about 3.65 times higher than that of the control after 3 h of low temperature stress. The data of expression under low temperature stress showed that Va Corr 413-PM2An-PM1X1 and -PM2B were down-regulated in response to hypothermia, and the expression range was lower than that of the corresponding genes in European grape, which were clustered in a cluster of Va Cor413-PM2AM-PM1X1 and -PM2B. In another cluster, the response of Va COR413-IM1 to hypothermia stress (24 h) was induced and expressed significantly. The results provided a reference basis for exploring the possible function and mechanism of the gene family members in response to cold stress. At the same time, it provides a theoretical basis for the molecular improvement of cold (freezing) resistance of grape plants.
【作者单位】： 宁夏大学农学院/葡萄与葡萄酒教育部工程研究中心/宁夏葡萄与葡萄酒研究院/宁夏葡萄与葡萄酒工程技术研究中心;
【基金】：国家自然科学基金地区科学基金项目(No.31560550) 园艺学“十三五”自治区优势特色学科建设(2015)110
【分类号】：S663.1
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本文编号：1856203