【摘要】：為了研究葡萄肌醇半乳糖苷合成酶基因家族的特性,以抗低溫品種左優(yōu)紅組培苗為材料,通過同源克隆技術(shù),獲得VvGolS2、VvGolS3和VvGolS4全長CDS,對其序列進(jìn)行了生物信息學(xué)分析,同時利用實(shí)時定量PCR技術(shù)探究了3個基因的組織表達(dá)特性和響應(yīng)逆境因子及相關(guān)信號分子的特性。結(jié)果顯示:3個基因c DNA大小分別為954,978,1 011 bp,分別編碼317,325,336個氨基酸序列;3個蛋白分子量分別為36.65,36.97,38.12 k Da;等電點(diǎn)分別為5.12,5.33和5.16,且均在C端含有一個疏水的五肽APXAA;實(shí)時熒光定量PCR證明,3個GolSs在葡萄不同組織中表達(dá)情況不同,VvGolS2和VvGolS4在葉片中表達(dá)量最高,而VvGolS3則在花和卷須中表達(dá)量最高;不同逆境因子及逆境信號分子均會影響VvGolS2、VvGolS3和VvGolS4基因的表達(dá)量,其中VvGolS2和VvGolS4基因的表達(dá)受鹽誘導(dǎo)最明顯,VvGolS3基因的表達(dá)受低溫誘導(dǎo)最明顯;另外,脅迫信號分子乙烯、過氧化氫和脫落酸均可誘導(dǎo)VvGolS2基因表達(dá)量上升,而VvGolS3基因的表達(dá)受脫落酸和乙烯的誘導(dǎo),乙烯、過氧化氫和硫化氫則均可誘導(dǎo)VvGolS4基因表達(dá)量上升。綜上推測,3個Vv GolSs均與葡萄抵御逆境脅迫相關(guān),但各自功能可能有所不同。
[Abstract]:In order to study the characteristics of Grape inositol galactoside synthase gene family, we obtained VvGolS2, VvGolS3 and VvGolS4 full-length CDSs by homologous cloning technique, and analyzed their sequences by bioinformatics. At the same time, the characteristics of tissue expression, response to stress factors and signal molecules of three genes were investigated by real-time quantitative PCR. The results showed that the c DNA of the three genes were 954, 978, 1011 BP, respectively, encoding 317325336 amino acid sequences, the molecular weight of three proteins were 36.65, 36.97, 38.12k Da., the isoelectric points were 5.125.33 and 5.16, and all contained a hydrophobic pentapeptide APXAAs at the C-terminal. Quantitative PCR showed that the expression of VvGolS2 and VvGolS4 in different tissues of grape was the highest, and the expression of VvGolS2 and VvGolS4 was the highest in different tissues of grape. The expression of VvGolS2 and VvGolS3 and VvGolS4 genes were affected by different stress factors and stress signal molecules, among which the expression of VvGolS2 and VvGolS4 genes was the most obvious induced by salt, and the expression of VvGolS3 gene was the most obvious induced by hypothermia. Stress signaling molecules ethylene, hydrogen peroxide and abscisic acid could induce the increase of VvGolS2 gene expression, while the expression of VvGolS3 gene was induced by abscisic acid and ethylene. Ethylene, hydrogen peroxide and hydrogen sulfide could induce the increase of VvGolS4 gene expression. It was suggested that all of the three VV GolSs were related to the resistance of grape to stress, but their functions might be different.
【作者單位】： 青島農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院山東省高校植物生物技術(shù)重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金項(xiàng)目(31572107;31540090;31401844)
【分類號】：S663.1

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