摘要
查尔酮合成酶及其异构酶是黄酮类化合物合成途径中的2个关键酶,为研究其在紫参薯花青素合成途径中的功能,利用RT-PCR技术从紫参薯Da56块茎中克隆到查尔酮合成酶(chalcone synthase,CHS)和查尔酮异构酶(chalcone isomerase,CHI)基因,分别命名为DaCHS和DaCHI。结果表明:DaCHS基因包含825 bp的完整开放阅读框(ORF),预测编码274个氨基酸;DaCHI基因包含663 bp的完整开放阅读框,预测编码221个氨基酸。生物信息学分析结果表明,紫参薯DaCHS与油棕Eg CHS亲缘关系最近,氨基酸相似性达到90%;紫参薯DaCHI与野茶树CaCHI亲缘关系最近,氨基酸相似性达到67%。实时荧光定量PCR检测结果表明,DaCHS和DaCHI基因均具有组织表达特异性,其中DaCHS在花中相对表达量最高,而DaCHI在块茎中相对表达量最高;在紫参薯块茎发育不同时期的表达分析表明,DaCHS、DaCHI均在薯块膨大期相对表达量最高。本研究通过对DaCHS和DaCHI基因全长cDNA序列的克隆与分析,为紫参薯花青素合成途径的遗传调控及花青素形成机理的研究奠定基础。
Chalcone synthase (CHS) and Chalcone isomerase (CHI) play an important role in the process of flavonoids synthesis. To investigate the function of the DaCHS and DaCHI gene in anthocyanin biosynthesis pathway of Dioscorea alata. L., the complete genes of ehaleone synthase(CHS) and ehaleone isomerase(CHl) were isolated from the turber of D. alata, by reverse transeription-polymerase chain reaction (RT-PCR) named as DaCHS and DaCHI. Sequencing results showed that the eDNA sequence of DaCHS gene contained an open reading frame (ORF) of 825 bp encoding a protenin of 274 amino acids, DaCHI gene contained an open reading frame (ORF) of 663 bp encoding a protenin of 221 amino acids. Phylogeneticanalyses showed that DaCHS and DaCHI had high identity to CHS and CHI protein of many plants. The highly similarity of DaCHS protenin was EgCHS protenin with 90% amino identity. The highly similarity of DaCHS protenin was CaCHI protenin with 67% amino identity. Quantitative real-time PCR analysis showed that the DaCHS and DaCHI expression had tissue specificity. The relative expression level of flowers were the highest in DaCHS. In DaCHI, the tubers had the hightest relative expression level. That expression analysis in different development periods of D. alata. tuber, DaCHS and DaCHI all had the hightest relative expression in expansion period of tubers.The cloning and analysis of full-length eDNA sequences of DaCHS and DaCHI genes should lay a foundation for the genetic regulation of anthoeyanin biosynthesis pathway and the mechanism of anthocyanin formation.
作者
赵景梅
黄东益
张青
许云
ZHAO Jingmei;HUANG Dongyi;ZHANG Qing;XU Yun(Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, Chin)
出处
《热带作物学报》
CSCD
北大核心
2018年第5期920-925,共6页
Chinese Journal of Tropical Crops
基金
海南省重点研发计划项目(No.ZDYF2016205)
国家自然科学基金(No.31260346)