葡萄HCT基因家族鉴定及其对低温胁迫的响应  

Identification of the Grape HCT Gene Family and Their Responses to Low-temperature Stress

作　　者：颜伟 陈慧婷 叶青[1] 刘广超 刘新[1] 侯丽霞[1] YAN Wei;CHEN Hui-ting;YE Qing;LIU Guang-chao;LIU Xin;HOU Li-xia(Key Lab of Plant Biotechnology in Universities of Shandong Province,College of Life Science,Qingdao Agricultural University,Qingdao 266109)

机构地区：[1]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,青岛266109

出　　处：《生物技术通报》2025年第2期175-186,共12页Biotechnology Bulletin

基　　金：山东省自然科学基金面上项目(ZR2023MC074);青岛市自然科学基金原创探索项目(23-2-1-178-zyyd-jch);山东省重点研发计划项目(2022LZGC0Y019)。

摘　　要：【目的】羟基肉桂酰转移酶(hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl transferase,HCT)是双子叶植物中催化绿原酸合成的关键酶,广泛参与植物非生物胁迫应答。在葡萄全基因组中鉴定VvHCT基因家族成员,分析相关基因低温胁迫下表达模式,为后续研究提供理论基础。【方法】基于葡萄全基因组信息,利用生物信息学方法对VvHCT家族进行鉴定,并对家族成员理化性质、染色体分布、基因结构、蛋白保守结构、系统进化、启动子顺式作用元件以及组织表达特性进行分析,进一步利用荧光定量PCR检测VvHCT基因在低温胁迫下表达模式,最后通过瞬时转化方法对VvHCT8功能进行验证。【结果】在葡萄全基因组中鉴定到VvHCT的11个家族成员,编码398-457个氨基酸,分布于4条染色体上;按系统发育特征分为3个亚族;启动子顺式作用元件分析显示,VvHCT家族成员启动子上含有光响应元件、激素响应元件、应激响应元件、组织特异性元件和昼夜节律响应元件等;不同VvHCT家族成员表达模式存在较大差异;低温胁迫处理后VvHCT5表达量下调,其他10个家族成员表达量均上调;瞬时过表达葡萄叶片结果显示,VvHCT8能通过降低丙二醛和活性氧含量,缓解低温对葡萄叶片叶绿素和细胞膜的损伤。【结论】VvHCT8参与葡萄抵御低温过程,可作为葡萄抗寒候选基因。【Objective】Hydroxycinnamoyl-CoA shikimate hydroxycinnamoyl transferase(HCT)is a key enzyme for catalyzing chlorogenic acid synthesis in dicotyledonous plants and is widely involved in plant abiotic stress response.We identified members of the VvHCT gene family in the whole grape genome and analyzed the expression pattern of the related genes under low-temperature stress,which may provide a theoretical basis for subsequent studies.【Method】Based on the whole genome information of grapevine,bioinformatics methods was sued to identify the VvHCT family,and analyze the physical and chemical properties,chromosome distribution,gene structure,protein conserved structure,systematic evolution,promoter cis-acting elements and tissue expression characteristics of the family members.Fluorescence quantitative PCR was adapted to detect the expression pattern of VvHCT gene under low-temperature stress,and transient transformation method to verify the function of VvHCT8 gene.【Result】Eleven family members of VvHCT were identified in the whole genome of grapevine,encoding 398-457 amino acids,distributed on four chromosomes.They were classified into three subfamilies according to their phylogenetic characteristics.The analysis of the promoter cis-acting elements showed that the promoters of the VvHCT family members contained light-responsive elements,hormone-responsive elements,stress-responsive elements,tissue-specific elements and circadian response elements.The expression patterns of different VvHCT family members differed greatly.The expression of VvHCT5 was down-regulated after lowtemperature stress treatment,while the expressions of the other 10 family members were up-regulated;the transient overexpression results of grape leaves showed that VvHCT8 alleviated the damages to chlorophyll and cell membranes of grape leaves by reducing the content of MDA and ROS.【Conclusion】VvHCT8 is involved in the process of low-temperature resistance in grapes and can be used as a candidate gene for cold resistance in grapes.

关 键 词：葡萄 VvHCT基因家族 生物信息学 低温 

分 类 号：Q943.2[生物学—植物学] S663.1[农业科学—果树学]