七彩虹竹4-香豆酸辅酶A连接酶基因(Ih4CL)的克隆及功能分析

Cloning and Functional Analysis of 4 Coumarate: Coenzyme A Ligase Gene from Indosasa hispida cv. ‘Rainbow'

4-香豆酸辅酶A连接酶(4CL)是苯丙烷类代谢途径的关键酶,在上游调控木质素代谢、黄酮代谢。本研究根据转录组数据设计的1对特异引物,采用逆转录PCR(RT-PCR)方法从七彩红竹1年生红秆中克隆到4-香豆酸辅酶A连接酶基因Ih4CL,该基因包含1 677bp完整的c DNA开放阅读框,编码558个氨基酸;生物信息学方法对Ih4CL编码的氨基酸蛋白序列进行的分析表明,Ih4CL与禾本科植物4CL蛋白具有较近的亲缘关系,其中与短花药野生稻的同源性达88%;荧光定量PCR检测结果显示Ih4CL基因在七彩红竹的嫩叶中的表达量明显低于茎秆中的表达量。

4-Coumarate coenzyme A ligase （4CL） is one of the key enzymes in the phenylpropanoid synthesis pathway, which produces several important secondary metabolites, such as flavonoids and lignins. In this study, a 4CL gene （ designated as Ih4CL） was obtained from the red culm of lndosasa hispida cv. ‘ Rainbow＇ by PCR with a pair of special primers designed based on the transcriptome date. The Ih4CL gene has a 1677bp open reading frame （ORF） that is predicted to encode a protein with 558 amino acids. Phylogenetic analysis showed that the Indosasa hispida cv. ‘Rainbow＇have more closer relative relationship with Poaceae plants and Ih4CL sequence shared high sequence identity （88%） with Oryza brachyantha. Real time PCR analysis showed that Ih4CL was most strongly expressed in the young white culm.