葡萄VvIQD10果形相关基因定位及其互作研究
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国家自然科学基金(31972384,31901975);


Study on Gene Location and Its Interaction of VvIQD10 Related to Grape Fruit Shape
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    摘要:

    以葡萄6个不同果形品种盛花期花序为材料,通过荧光定量PCR方法分析比较葡萄IQ67结构域(IQ67 domain, IQD)基因家族成员VvIQD10的表达;从椭圆形葡萄品种‘天山’中克隆出VvIQD10基因编码区序列,对其进行生物信息学分析,采用亚细胞定位瞬时表达载体转化烟草的方法研究VvIQD10蛋白在细胞中的分布情况,并通过酵母双杂交实验和双分子荧光互补试验进行蛋白互作研究。结果表明:(1)VvIQD10基因在长果形葡萄品种中的表达量高于近圆形葡萄品种。(2)VvIQD10基因开放阅读框长度为1 401 bp,编码466个氨基酸。(3)VvIQD10蛋白为不稳定亲水性蛋白,无信号肽和跨膜区,但含保守的IQ67结构域,主要结构为α螺旋和随机卷曲,与猕猴桃IQD家族成员(PSS09955.1)亲缘关系最为接近。(4)VvIQD10蛋白主要定位于微管和质膜,并且直接与细胞骨架组织相关蛋白——钙调素类蛋白(VvCML)相互作用。研究认为,VvIQD10基因可能参与葡萄果形调控,葡萄VvCML蛋白从胞质到微管的募集依赖于VvIQD10,推测VvIQD10蛋白可能通过和钙调素类蛋白相结合来调控细胞骨架运动,进而参与葡萄果实形状的变化。

    Abstract:

    Present study used the inflorescences of six grape varieties with different fruit shapes as the experimental materials. The expression of VvIQD10, a member of the grape IQ67 domain (IQD) gene family, was compared by fluorescence quantitative PCR analysis. The coding region sequence of the VvIQD10 gene was cloned from the oval grape variety ‘Tianshan’ and analyzed by bioinformatics. The subcellular location transient expression vector was used to transform tobacco to study the distribution of VvIQD10 protein in the cell. Yeast2hybrid assay and bimolecular fluorescence complementation test were used to study protein interaction. The results showed that: (1) the expression level of VvIQD10 gene in longshaped grape varieties was higher than that of nearly round grape varieties. (2) The length of the open reading frame of the VvIQD10 gene was 1 401 bp, which encoded 466 amino acids. (3) VvIQD10 protein was an unstable hydrophilic protein with no signal peptide and transmembrane domain, but it contained a conserved IQ67 domain. The main structure was alpha helix and random coil. It was closely related to a member of the kiwi IQD family (PSS09955.1). (4) VvIQD10 protein is mainly located in microtubules and plasma membranes, and directly interacted with cytoskeletal tissuerelated proteinscalmodulinlike proteins (VvCML). Studies suggested that the VvIQD10 gene may be involved in the regulation of grape fruit shape, and the recruitment of VvCML protein from cytoplasm to microtubules depended on VvIQD10. It is speculated that VvIQD10 protein may regulate cytoskeletal movement by combining with calmodulinlike proteins, and then participate in shaping of grape berries.

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余 欣,董 阳,刘 静,等.葡萄VvIQD10果形相关基因定位及其互作研究[J].西北植物学报,2021,41(6):899-909

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  • 在线发布日期: 2021-07-20
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