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.
Zhongguo Zhong Yao Za Zhi. 2024 Jan;49(2):361-369. doi: 10.19540/j.cnki.cjcmm.20230902.103.
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.
4-香豆酸:辅酶A连接酶(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中的关键酶。在本研究中,以拟南芥的13个4CL家族成员作为参考序列,从已报道的菘蓝基因组中鉴定菘蓝4CL基因家族候选成员。进一步进行了生物信息学分析以及4CL基因表达模式和类黄酮积累模式的分析。获得了13个4CL基因,命名为Ii4CL1 - Ii4CL13,它们分布在第1、2、3、4和6号染色体上。基因结构和保守结构域分析表明,菘蓝4CL基因的内含子数量在1至12之间,蛋白质结构域高度保守。顺式作用元件分析表明,菘蓝4CL基因家族启动子序列中存在多个响应元件,其中茉莉酸的反应元件数量最多。共线性分析表明,菘蓝和拟南芥的4CL基因家族成员之间关系密切。qPCR结果显示,4CL基因家族的表达分析表明,10个4CL基因在菘蓝地上部分具有较高的表达水平。菘蓝不同部位类黄酮含量测定表明,类黄酮主要积累在植物地上部分。本研究为进一步探究4CL基因家族在菘蓝类黄酮生物合成中的作用提供了依据。