Cloning and Expression Analysis of NF‑YB3 Gene from Vitis vinifera

doi:10.15933/j.cnki.1004-3268.2024.04.012

Abstract

Abstract: The full length of the VvNF‑YB3 gene was cloned from the European grape‘Thompson Seedless’by PCR,to explore the function of the VvNF‑YB3 gene in grape growth and development and response to abiotic stresses.The coding region sequence(CDS)of the VvNF‑YB3 gene was 624 bp in length,encoding 207 amino acids,with an isoelectric point of 6.31，an instability index of 27.66,and a total average hydrophobic index of -0.738.Therefore,VvNF⁃YB3 was an acidic and stable hydrophilic protein.Through bioinformatics analysis,it was found that the VvNF‑YB3 gene was located on chromosome 10 of the European grape,containing an intron and a CBFD_NFYB_HMF conserved domain,and showed the highest identity with VrNF⁃YB3 from Vitis riparia.The prediction result of secondary structure showed that VvNF⁃YB3 was mainly composed of α⁃helix and random coil.Correspondingly,the tertiary structure was a monomeric helix⁃coil⁃helix.The analysis of the cis⁃acting elements of the VvNF‑YB3 gene promoter suggested that it might respond to hormones and stresses.To prove this inference,its expression patterns under different treatments were analyzed.The VvNF‑YB3 gene was expressed in leaves,tendrils,flowers,young fruits,and stems of Red Globe,and the expression level was the highest in mature leaves.PEG simulated drought,low⁃temperature,and high⁃temperature stresses could change the expression level of the VvNF‑YB3 gene,and VvNF‑YB3 was a stress responsive gene.In addition,most of the interacting proteins of VvNF⁃YB3 were subunits of the nuclear factor Y family,and there were 27 transcription factors such as ERF,bHLH,ARF,etc.in the upstream of the VvNF‑YB3 gene.It is speculated that the VvNF‑YB3 gene may respond to various environmental stresses by being regulated by upstream transcription factors and interacting with other NF⁃Y proteins.

Key words: Grape, NF?YB3, Gene cloning, Bioinformatics analysis, Expression analysis

摘要： 为探究VvNF-YB3基因在葡萄生长发育和响应非生物胁迫过程中的功能，以无核白为试材，克隆得到了VvNF-YB3 基因编码区序列，全长为624 bp，编码207个氨基酸，蛋白质等电点6.31，不稳定指数27.66，总平均疏水指数-0.738，为一酸性、稳定的亲水蛋白。通过生物信息学分析发现，VvNF-YB3基因位于10号染色体，含有1个内含子，编码蛋白质存在1个CBFD_NFYB_HMF结构域，与河岸葡萄NFYB3相似性最高。VvNF-YB3的二级结构中α-螺旋和无规则卷曲占比较高，三级结构为单体的螺旋-卷曲-螺旋，启动子中包含响应激素和胁迫的多个顺式作用元件。VvNF-YB3 基因在葡萄的叶片、卷须、花、幼果和茎中均表达，且在成熟的叶片中表达量最高。PEG模拟的干旱、低温和高温胁迫均可影响VvNF-YB3基因的表达，VvNF-YB3为一胁迫响应基因。VvNF-YB3的互作蛋白大多是核因子Y家族的亚基，ERF、bHLH、ARF等27个转录因子可能调控VvNF-YB3 基因的表达。由此推测，VvNF-YB3 基因可能通过受到上游转录因子的调控并与其他NF-Y蛋白互作响应各种环境胁迫。

关键词: 葡萄, NF?YB3, 基因克隆, 生物信息学分析, 表达分析

CLC Number:

S663.1

CUI Xiaoyue, SHANG Hongquan, LÜ Zhongwei, LOU Yusui, ZHANG Ke, FAN Hongjie, WU Wenying, ZHANG Xiaofeng. Cloning and Expression Analysis of NF‑YB3 Gene from Vitis vinifera[J]. Journal of Henan Agricultural Sciences, 2024, 53(4): 111-118.

崔小月, 尚泓泉, 吕中伟, 娄玉穗, 张柯, 樊红杰, 吴文莹, 张晓锋. 欧洲葡萄NF-YB3 基因克隆与表达分析[J]. 河南农业科学, 2024, 53(4): 111-118.

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