菘蓝4-香豆酸:辅酶A连接酶全基因家族的鉴定及表达分析  

Identification and expression analysis of whole gene family of Isatis indigotica 4-coumarate:CoA ligase

作　　者：张苛苛 谭宇萍 徐欢欢 孙术富 肖建才 徐朝阳 王晨晨 朱哈修 杨健[1] 李大勇 唐金富[1] ZHANG Ke-ke;TAN Yu-ping;XU Huan-huan;SUN Shu-fu;XIAO Jian-cai;XU Zhao-yang;WANG Chen-chen;ZHU Ha-xiu;YANG Jian;LI Da-yong;TANG Jin-fu(State Key Laboratory for Quality Assurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Beijing Key Laboratory of Vegetable Germplasm Improvement,Beijing Vegetable Research Center,Beijing Academy of Agricultural and Forestry Sciences,Beijing 100097,China;College of Life Sciences,Shandong Normal University,Ji′nan 250014,China)

机构地区：[1]中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室,北京100700 [2]北京市农林科学院蔬菜研究所蔬菜种质改良北京市重点实验室,北京100097 [3]山东师范大学生命科学学院,山东济南250014

出　　处：《中国中药杂志》2024年第2期361-369,共9页China Journal of Chinese Materia Medica

基　　金：中国中医科学院科技创新工程项目(CI2021A04114);国家自然科学基金项目(81974517)。

摘　　要：4-香豆酸:辅酶A连接酶(4-coumarate:CoA ligase,4CL)是植物中类黄酮、可溶性酚酯、木脂素和木质素等苯丙烷类化合物上游途径的关键酶。该研究基于菘蓝基因组数据库,以拟南芥Arabidopsis thaliana的13个4CL家族成员为参考序列,获取菘蓝4CL基因家族的候选成员,并进行生物信息学分析、4CL基因表达模式及类黄酮代谢成分积累模式等研究。结果共获得13条菘蓝4CL基因家族成员,分别命名为Ii4CL1~Ii4CL13,分别定位到1、2、3、4、6号染色体上。基因结构与保守结构域分析发现菘蓝4CL基因家族成员的内含子数目为1~12,蛋白结构域具有高度保守性。顺式作用元件分析表明在菘蓝4CL基因家族成员启动子序列中存在多个响应元件,其中茉莉酸反应元件数量最多。共线性分析表明菘蓝4CL基因家族成员与同科植物拟南芥之间存在更近的亲缘关系。qPCR结果显示,菘蓝4CL基因家族表达分析发现10个4CL基因在菘蓝的地上部分相对表达量更高。对菘蓝不同部位类黄酮化合物的含量进行检测,发现菘蓝的类黄酮主要积累在地上部分。该研究为深入探索菘蓝4CL基因家族成员参与菘蓝类黄酮生物合成的分子机制奠定了基础。The 4-coumarate:CoA ligase(4CL) is a key enzyme in the upstream pathway of phenylpropanoids such as flavonoids,soluble phenolic esters,lignans,and lignins in plants.In this study,13 4CL family members of Arabidopsis thaliana were used as reference sequences to identify the 4CL gene family candidate members of Isatis indigotica from the reported I.indigotica genome.Further bioinformatics analysis and analysis of the expression pattern of 4CL genes and the accumulation pattern of flavonoids were carried out.Thirteen 4CL genes were obtained,named Ii4CL1-Ii4CL13,which were distributed on chromosomes 1,2,3,4,and 6.The analysis of the gene structure and conserved structural domains revealed the intron number of I.indigotica 4CL genes was between 1 and 12 and the protein structural domains were highly conserved.Cis-acting element analysis showed that there were multiple response elements in the promoter sequence of I.indigotica 4CL gene family,and jasmonic acid had the largest number of reaction elements.The collinearity analysis showed that there was a close relationship between the 4CL gene family members of I.indigotica and A.thaliana.As revealed by qPCR results,the expression analysis of the 4CL gene family showed that 10 4CL genes had higher expression levels in the aboveground part of I.indigotica.The content assay of flavonoids in different parts of I.indigotica showed that flavonoids were mainly accumulated in the aboveground part of plants.This study provides a basis for further investigating the roles of the 4CL gene family involved in the biosynthesis of flavonoids in I.indigotica.

关 键 词：4-香豆酸:辅酶A连接酶 菘蓝 生物信息学分析 类黄酮 

分 类 号：S567.2[农业科学—中草药栽培]