刺葡萄白藜芦醇合成酶基因对不同光质的响应及转录因子筛选
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福建省人民政府中国农业科学院农业高质量发展超越 “5511” 项目(XTCXGC2021014);


Response of Resveratrol Synthase Gene (RS1) to Different Light Quality and Transcription Factor Screening in Vitis davidii
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    摘要:

    为探究白藜芦醇合成酶基因(RS)的表达模式和转录调控特征,该研究以刺葡萄愈伤组织为材料,采用RTPCR方法进行RS1基因克隆,分析RS1基因在8种不同光质培养条件下的表达模式,并对靶向RS1的转录因子进行预测分析和筛选验证。结果表明:(1)成功从刺葡萄中克隆获得RS1基因(GenBank登录号为OM339527);RS1基因开放阅读框为1 179 bp,由2个外显子和1个内含子组成,编码392个氨基酸,为亲水性无信号肽的细胞质定位蛋白,磷酸化修饰主要发生于苏氨酸和丝氨酸位点上,蛋白质的二级结构主要由α螺旋、无规则卷曲和延伸链组成。(2)RS1启动子具有多个光响应和转录因子识别与结合元件,还涉及激素调控、生长发育、环境条件响应。(3)转录调控预测发现,靶向RS1的转录因子来自9个家族,共有23个成员,其中MYBDof基因家族具有多个成员和RS1启动子结合位点。(4)共线性分析表明,葡萄与毛果杨的共线性最高,MYBDIVDof5.1具有多个共线基因。(5)转录水平分析显示,长波光促进RS1的表达,在不同光质和培养阶段下MYBDIV与靶基因RS1具有相同的表达模式,而Dof5.1的表达模式与RS1呈相反趋势,表明MYBDIV和Dof5.1分别通过正负调控参与RS1的转录表达。

    Abstract:

    To explore the expression pattern and transcriptional regulation characteristics of resveratrol synthase gene (RS), we cloned RS1 gene using RTPCR from Vitis davidii callus. The expression pattern of RS1 was studied under 8 different light qualities, and the transcription factors targeting at RS1 were predicted and verified. The results showed that: (1) RS1 gene (GenBank accession number: OM339527) was cloned from V. davidii. The open reading frame of RS1 gene was 1 179 bp composed of 2 exons and 1 intron, encoding 392 amino acids. RS1 protein was a hydrophilic protein without signal peptide, and was predicted to be cytoplasmic localization. Phosphorylation modification mainly occurred at threonine and serine sites. The secondary structure of protein mainly consists of αhelix, random coil and strand. (2) There were multiple light response and transcription factor recognition/binding elements in RS1 promoter, other cisactive elements were involved in hormone regulation, growth and development, and environmental response. (3) Twentythree transcription factors belonging to 9 families targeting at RS1 were predicted by transcriptional regulation analysis, among which, MYB and Dof had many family members and RS1 promoter binding sites. (4) Collinearity analysis showed that grapevine had the highest collinearity with Populus trichocarpa, MYBDIV and Dof5.1 genes had multiple collinear genes. (5) Relative qPCR analysis indicated that longwavelength light quality promoted RS1 expression. MYBDIV and targeted gene RS1 had the same expression pattern under different light qualities and culture stages, while Dof5.1 revealed contrary results with RS1. This research suggests MYBDIV and Dof5.1 were involved in the biosynthesis of resveratrol by positively and negatively regulating the expression of RS1, respectively.

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赖恭梯,阙秋霞,赖谱富,等.刺葡萄白藜芦醇合成酶基因对不同光质的响应及转录因子筛选[J].西北植物学报,2022,42(7):1113-1123

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  • 在线发布日期: 2022-08-22
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