食品科学 ›› 2024, Vol. 45 ›› Issue (12): 109-115.doi: 10.7506/spkx1002-6630-20230726-292

• 生物工程 • 上一篇    

绿豆7S球蛋白基因鉴定及在籽粒发育中的表达分析

丁冬会,张雨朋,周人玲,席武洋,耿旎周,常玮,赵青平,刘嘉斐,杨树琼   

  1. (南阳师范学院生命科学与农业工程学院,河南 南阳 473061)
  • 发布日期:2024-07-10
  • 基金资助:
    国家自然科学基金青年科学基金项目(32100225);河南省自然科学基金青年项目(232300420213); 河南省重点研发与推广专项(科技攻关)项目(222102110275);南阳市科技计划项目(KJGG014)

Identification of 7S Globulin Genes in Mung Bean (Vigna radiata L.) and Their Expression Analysis during Seed Development

DING Donghui, ZHANG Yupeng, ZHOU Renling, XI Wuyang, GENG Nizhou, CHANG Wei, ZHAO Qingping, LIU Jiafei, YANG Shuqiong   

  1. (College of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang 473061, China)
  • Published:2024-07-10

摘要: 为阐明绿豆7S(Vigna radiata 7S,Vr7S)球蛋白基因的基本性质及其在籽粒生长发育过程中的表达模式,本研究运用生物信息学、反转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)对10 个Vr7S球蛋白基因的理化性质、基因结构特征、蛋白三级结构、系统发育关系及基因表达谱等进行研究。结果表明,Vr7S球蛋白基因编码的蛋白长度为246~594 个氨基酸,等电点位于5.02~6.40之间,分子质量为27.82~70.02 kDa;所有Vr7S球蛋白基因包含3~7 个外显子、2~6 个内含子、5~8 个保守基序;除Vr7S B1具有1 个cupin保守结构域外,其余9 个基因均含有2 个cupin结构域;Vr7S球蛋白三级结构有4 种类型,主要包含α-螺旋、β-链等结构;在cupin保守结构域中,绿豆和其他物种共有67 个保守氨基酸残基,绿豆与赤豆7S球蛋白基因间的同源关系最近,其次是菜豆、木豆及大豆等。基于不同绿豆品种、不同发育时期籽粒的转录组测序和RT-PCR表达谱分析显示,Vr7S球蛋白基因伴随着绿豆籽粒的发育成熟呈现表达量升高的趋势,这与其他豆科物种7S球蛋白基因的表达模式一致,表明其在绿豆籽粒贮藏蛋白的生成调控过程中具有积极作用。本研究为进一步解析Vr7S球蛋白基因在绿豆籽粒发育中的生物学功能奠定了基础。

关键词: 绿豆;7S球蛋白;籽粒发育;基因鉴定;表达谱分析

Abstract: In order to elucidate the basic properties of Vigna radiata 7S (Vr7S) globulin genes and their expression patterns during seed growth and development, the physicochemical properties, structural characteristics, protein tertiary structures, phylogenetic relationship and expression profiles of 10 Vr7S globulin genes were studied using bioinformatics and reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the proteins encoded by the Vr7S globulin genes consisted of 246 to 594 amino acid residues, with isoelectric point (pI) ranging from 5.02 to 6.40, and molecular mass ranging from 27.82 to 70.02 kDa. All Vr7S globulin genes contained 3–7 exons, 2–6 introns, and 5–8 conserved motifs. Vr7S B1 had one conserved cupin domain, while the other nine genes contained two cupin domains. Vr7S globulin had four types of tertiary structures including α-helix and β-chain. In the conserved cupin domain, 67 conserved amino acid residues were shared by mung bean and other species, and mung bean 7S globulin genes had the closest homology adzuki bean 7S globulin genes followed by those of French bean, pigeon pea and soybean. The RNA sequencing (RNA-Seq) and RT-PCR expression profiles of mung bean seeds at different development stages showed that the expression level of the Vr7S globulin genes increased with the maturity of mung bean seeds, which was consistent with the expression pattern of 7S globulin genes in other legume species. These results indicated that the Vr7S globulin genes played a positive role in regulating the generation of mung bean seed storage protein. This study may lay a foundation for further analyzing the biological function of the Vr7S globulin genes in the seed development of mung bean.

Key words: mung bean; 7S globulin; seed development; gene identification; expression profile analysis

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