本文選題：抗寒性 + 冷適應(yīng)蛋白基因(COR)��； 參考：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2017年03期

【摘要】：冷適應(yīng)蛋白(cold-regulated,COR413)基因作為植物特有的一類低溫脅迫響應(yīng)功能基因,在抗寒過程中發(fā)揮重要作用。為了鑒定葡萄屬(Vitis)植物COR413基因參與低溫脅迫響應(yīng)的功能特點(diǎn),本研究基于歐洲葡萄基因組數(shù)據(jù)庫(kù)挖掘,采用同源克隆法獲得了抗寒種質(zhì)山葡萄(Vitis amurensis)COR413基因家族成員的4條序列,分別命名為Va COR413-PM2A、-PM1X1、-IM1與-PM2B。經(jīng)序列分析表明,所克隆的4條山葡萄COR413基因家族成員與歐洲葡萄相應(yīng)基因的核苷酸與氨基酸序列高度同源,其核苷酸序列的差異僅為個(gè)別單核苷酸的替代。所克隆的4個(gè)山葡萄冷適應(yīng)蛋白家族基因均含有WCOR413保守結(jié)構(gòu)域、跨膜結(jié)構(gòu)域與絲/蘇/酪氨酸磷酸化位點(diǎn),其中僅Va COR413-PM1X1含有一個(gè)預(yù)測(cè)的N端信號(hào)肽序列,僅Va COR413-PM2B包含1個(gè)糖基化磷脂酰肌醇錨點(diǎn)。源于2個(gè)葡萄種質(zhì)中COR413基因家族成員可以聚類為2個(gè)明顯的簇,COR413-PM2A,-PM1X1與-PM2B聚類在一簇,而COR413-IM1分布在另外一簇。半定量RT-PCR組織特異性檢測(cè)結(jié)果表明,Va COR413-PM2A、-PM1X1、-IM1與-PM2B在山葡萄不同組織和器官中表達(dá)存在差異,且不同于歐洲葡萄相應(yīng)基因的表達(dá)模式;其中山葡萄Va COR413-PM2A、-PM1X1、-IM1與-PM2B在所有檢測(cè)的組織中均表達(dá)且強(qiáng)度不同,Va COR413-PM2A與-PM1X1在嫩梢、卷須、花絮以及幼葉中表達(dá)量較高。在低溫脅迫條件下,山葡萄COR413基因家族4個(gè)成員參與低溫脅迫的表達(dá)模式分為2類,而歐洲葡萄中相應(yīng)成員有3種模式;山葡萄Va COR413-IM1受低溫脅迫24 h表達(dá)量最高,為對(duì)照的9.57倍,而歐洲葡萄Vv COR413-IM1在低溫脅迫3 h后的表達(dá)量較高,約為對(duì)照的3.65倍。低溫脅迫下的表達(dá)數(shù)據(jù)表明,山葡萄COR413基因家族成員中聚類為一簇的Va COR413-PM2A,-PM1X1與-PM2B響應(yīng)低溫總體呈下調(diào)表達(dá),其表達(dá)幅度均低于歐洲葡萄相應(yīng)基因的表達(dá)量,而聚類在另外一簇的Va COR413-IM1對(duì)低溫脅迫(24 h)響應(yīng)誘導(dǎo)表達(dá)顯著;研究結(jié)果為探明該基因家族成員在葡萄響應(yīng)低溫脅迫過程中的可能功能和作用機(jī)制提供了參考依據(jù),同時(shí)為葡萄屬植物抗寒(凍)性分子改良提供了理論依據(jù)。
[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.
【作者單位】： 寧夏大學(xué)農(nóng)學(xué)院/葡萄與葡萄酒教育部工程研究中心/寧夏葡萄與葡萄酒研究院/寧夏葡萄與葡萄酒工程技術(shù)研究中心;
【基金】：國(guó)家自然科學(xué)基金地區(qū)科學(xué)基金項(xiàng)目(No.31560550) 園藝學(xué)“十三五”自治區(qū)優(yōu)勢(shì)特色學(xué)科建設(shè)(2015)110
【分類號(hào)】：S663.1
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本文編號(hào)：1856203