葡萄肌醇半乳糖苷合成酶基因VvGolS2-4的克隆及表达特性分析

发布时间：2018-08-11 16:38

【摘要】：为了研究葡萄肌醇半乳糖苷合成酶基因家族的特性,以抗低温品种左优红组培苗为材料,通过同源克隆技术,获得VvGolS2、VvGolS3和VvGolS4全长CDS,对其序列进行了生物信息学分析,同时利用实时定量PCR技术探究了3个基因的组织表达特性和响应逆境因子及相关信号分子的特性。结果显示:3个基因c DNA大小分别为954,978,1 011 bp,分别编码317,325,336个氨基酸序列;3个蛋白分子量分别为36.65,36.97,38.12 k Da;等电点分别为5.12,5.33和5.16,且均在C端含有一个疏水的五肽APXAA;实时荧光定量PCR证明,3个GolSs在葡萄不同组织中表达情况不同,VvGolS2和VvGolS4在叶片中表达量最高,而VvGolS3则在花和卷须中表达量最高;不同逆境因子及逆境信号分子均会影响VvGolS2、VvGolS3和VvGolS4基因的表达量,其中VvGolS2和VvGolS4基因的表达受盐诱导最明显,VvGolS3基因的表达受低温诱导最明显;另外,胁迫信号分子乙烯、过氧化氢和脱落酸均可诱导VvGolS2基因表达量上升,而VvGolS3基因的表达受脱落酸和乙烯的诱导,乙烯、过氧化氢和硫化氢则均可诱导VvGolS4基因表达量上升。综上推测,3个Vv GolSs均与葡萄抵御逆境胁迫相关,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.
【作者单位】： 青岛农业大学生命科学学院山东省高校植物生物技术重点实验室;
【基金】：国家自然科学基金项目(31572107;31540090;31401844)
【分类号】：S663.1

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