中国辣椒ADH基因家族鉴定及表达分析  

Identification and Expression Analysis of the ADH Gene Family in Chinese Pepper(Capsicum chinense)

作　　者：叶楚 林蔚 王川 谢炳春 徐小万[2] 徐晓美[2] 王恒明[2] 吕俊恒 邓明华[4] 吴智明[1] 李涛[2] 衡周 YE Chu;LIN Wei;WANG Chuan;XIE Bingchun;XU Xiaowan;XU Xiaomei;WANG Hengming;LYU Junheng;DENG Minghua;WU Zhiming;LI Tao;HENG Zhou(College of Horticulture and Landscape Architecture,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China;Vegetable Research Institute,Guangdong Academy of Agriculture Sciences/Guangdong Key Laboratory of New Technology Research of Vegetables,Guangzhou 510640,China;Guangdong Provincial Agricultural Technology Extension Center,Guangzhou 510500,China;College of Horticulture and Landscape Architecture,Yunnan Agriculture University,Kunming 650000,China)

机构地区：[1]仲恺农业工程学院园艺园林学院,广东广州510225 [2]广东省农业科学院蔬菜研究所/广东省蔬菜新技术研究重点实验室,广东广州510640 [3]广东省农业技术推广中心,广东广州510500 [4]云南农业大学园艺园林学院,云南昆明650000

出　　处：《广东农业科学》2025年第2期95-107,共13页Guangdong Agricultural Sciences

基　　金：国家重点研发计划项目(2023YFD1201502);广东省农业厅种业振兴与科技兴农项目(2024-440000-87020100-8753);广州市科技计划项目(2023B03J1082);毕节市科学技术揭榜挂帅项目。

摘　　要：[目的]中国辣椒(Capsicum chinense)为茄科(Solanaceae)辣椒属(Capsicum)植物,大多具由支链酯类物质产生的浓郁果香。乙醇脱氢酶(Alcohol dehydrogenase,ADH)是影响支链酯类物质含量的重要酶,催化相应的醇和醛之间的可逆转化。鉴定中国辣椒ADH基因家族成员并分析其在植株各部位的表达模式,可为辣椒果实香气合成机理研究提供参考。[方法]通过生物信息学方法、实时荧光定量PCR分析中国辣椒ADH基因家族成员及表达特征,再利用气相色谱质谱联用(Gaschromatography-mass spectrometry,GC-MS)方法分析中国辣椒果实挥发性物质。[结果]从中国辣椒基因组中鉴定到8个ADH基因,分布在4条染色体上,分别命名为CcADH1~8。理化性质分析结果显示,ADH基因家族编码的氨基酸序列长度在368~457 aa,分子质量范围为40~49 kD,等电点范围为5.50~9.12,CcADH4~6的亲水系数为正值、其余成员为负值;不稳定指数为22.23~44.81。蛋白质结构预测显示,CcADH的二级结构以α-螺旋和无规则卷曲为主,三级结构中CcADH4和CcADH6、CcADH7和CcADH8的结构相近。亚细胞定位预测发现,CcADH均定位于细胞质中。启动子区域顺式作用元件预测结果显示,CcADH启动子上存在12种顺式调控元件。时空表达分析表明,CcADH6在‘金田1号’‘7054’植株各器官均未检测到表达。在‘金田1号’中,CcADH7在花和果实第4时期高表达;在‘7054’中,CcADH5在果实第3时期高表达。利用GC-MS检测果实不同发育时期香气物质含量变化发现,‘金田1号’在果实第4时期香气物质总含量最高,而‘7054’在果实第3时期香气物质总含量最高。[结论]中国辣椒果实中的挥发性物质在其发育中后期大量积累,CcADH基因家族中的CcADH5和CcADH7分别与‘7054’和‘金田1号’果实中支链酯类物质变化趋势一致。它们参与的反应为下游的醇酰基转移酶(Alcohol acetyl transferase,AAT)提供特异底物,影响果实【Objective】Capsicum chinense is a member of the Solanaceae family and Capsicum genus,most of which have a rich fruity aroma produced by branched-chain esters.Alcohol dehydrogenase(ADH)is an important enzyme that affects the content of branched-chain esters,catalyzing the reversible transformation between corresponding alcohols and aldehydes.Identifying the members of the ADH gene family in C.chinense,and analyzing their expression patterns in different parts of the plant will provide references for the aroma synthesis mechanism of pepper fruit.【Method】Through bioinformatics analysis,real-time fluorescent quantitative PCR,and gaschromatography-mass spectrometry,the volatile substances and the ADH gene family in C.chinense were identified and analyzed for expression characteristics.【Result】Eight ADH genes were identified from the C.chinense genome,distributed across four chromosomes and named CcADH1-8.The prediction of physicochemical properties showed that lengths of the amino acid sequence encoded by the ADH gene family range from 368 to 457 aa,with molecular weights between 40 to 49 kD,and isoelectric points between 5.50 to 9.12.The hydrophilicity coefficients of CcADH4-6 were positive,while the rest of the gene family members were negative.The instability indices were ranged from 22.23 to 44.81.Protein structure prediction revealed that the secondary structure of CcADH was predominantly composed ofα-helices and random coils,while CcADH4 and CcADH6,as well as CcADH7 and CcADH8,were similar in the tertiary structurer.Subcellular localization prediction showed that all CcADHs were localized in the cytoplasm.The prediction results of cis-acting elements in the promoter region showed the promoters of CcADH contained 12 types of cis-regulatory elements.Spatiotemporal expression analysis showed that CcADH6 had no detectable expression in‘JT-1’and‘7054’Capsicum plants,CcADH7 was highly expressed in flowers and the 4th stage of fruit development in‘JT-1’,and CcADH5 was highly expressed in the 3r

关 键 词：中国辣椒 乙醇脱氢酶 支链醇 基因鉴定 表达分析 生物信息学 实时荧光定量PCR 

分 类 号：S641.3[农业科学—蔬菜学]