| 本文已被：浏览 1355次 下载 625次 |
|
|
|
| 马缨杜鹃查尔酮合酶的基因克隆及表达分析 |
|
胡荣美1, 舒合凤1, 朱灵1, 张习敏2, 龚记熠1,2, 孙威1,2*
|
|
( 1.贵州师范大学 生命科学学院/植物生理与发育调控重点实验室,贵阳 550025；2.西南喀斯特山地生物多样性保护重点实验室,贵阳 550025 )
|
|
| 摘要: |
| 为探究查尔酮合酶（Chalcone Synthase，CHS）在马缨杜鹃（Rhododendron delavayi）花色形成中的作用，该研究以马缨杜鹃为材料，克隆了RdCHS2编码区全长，并对其组织表达情况、体外催化活性及体内功能进行了解析。结果表明：（1）RdCHS2的CDS全长为1 170 bp，编码389个氨基酸，与其他植物CHS归为一类，具有CHS典型的保守基序。（2）表达分析显示，RdCHS2在花葶中表达最高，雄蕊中表达最低，伴随开花过程中表达量呈逐渐上升趋势。（3）酶活检测显示，RdCHS2可分别催化丙二单酰辅酶A（malonyl-CoA）与对香豆酰辅酶A（p-coumaroyl-CoA）生成柚皮素查尔酮（naringenin chalcone），催化丙二单酰辅酶A与肉桂酰辅酶A（cinnamoyl-CoA）生成松属素查尔酮（pinocembrin chalcone）。（4）突变体互补实验显示，RdCHS2可成功恢复拟南芥tt4突变体子叶与下胚轴中花青素苷的合成。该研究结果表明，RdCHS2具有查尔酮合酶活性，可以参与花青素苷的合成，为今后进一步明确该基因的具体功能奠定了基础。 |
| 关键词: 花青素苷，马缨杜鹃，查尔酮合酶，酶活检测，遗传转化 |
| DOI：10.11931/guihaia.gxzw202504046 |
| 分类号: |
| 基金项目:贵州省自然科学基金（ZK〔2023〕270）；贵州省高等学校高山杜鹃病虫害绿色防控重点实验室项目（黔教技〔2022〕044号）；贵州师范大学资助项目（[2021]B04）。 |
|
| Cloning and expression analysis of RdCHS2 from Rhododendron delavayi |
|
HU Rongmei1, SHU Hefeng1, ZHU Ling1, ZHANG Ximin2, GONG Jiyi1,2, SUN Wei1,2*
|
|
( 1 Key Laboratory of Plant Physiology and Development Regulation/School of Life Science, Guizhou Normal University, Guiyang 550025, China; 2 Key Laboratory of State Forestry Administration on Biodiversity Conservation in Karst Mountain Area of Southwest of China, Guiyang 550025, China )
|
| Abstract: |
| To elucidate the role of chalcone synthase (CHS) in flower color formation of Rhododendron delavayi, the full-length coding region of RdCHS2 was cloned using R. delavayi as materials, meanwhile, its expression profile, catalytic activity in vitro and biological function in vivo were analyzed. The results were as follows: (1) The full length CDS of RdCHS2 was 1170 bp encoding 389 amino acids. Phylogenetic analysis displayed that RdCHS2 was grouped into the same clade with CHS from other plants and had typical conserved motifs of CHS. (2) Gene expression analysis showed that the transcript of RdCHS2 was highest in scapes and lowest in stamens, and its expression level increased gradually during flowering. (3) Enzymatic assays confirmed that RdCHS2 catalyzed the synthesis of naringenin chalcone from malonyl-CoA and p-coumaroyl-CoA, as well as catalyzed the synthesis of pinocembrin chalcone from malonyl-CoA and cinnamoyl-CoA. (4) Complementation experiments in Arabidopsis thaliana tt4 mutant demonstrated that heterologous expression of RdCHS2 successfully restored anthocyanin biosynthesis in cotyledons and hypocotyls. These results show that RdCHS2 exhibits typical CHS activity and participates in the biosynthesis of anthocyanin, which lay a foundation for further clarifying the specific function of this gene in the future. |
| Key words: anthocyanin, Rhododendron delavayi, chalcone synthase, enzyme assay, genetic transformation |
