山茶属三个F3H基因的分子特性、系统进化及蛋白结构差异分析
发布时间：2019-07-25 15:16
【摘要】：黄烷酮3-羟化酶基因(flavanone 3-hydroxylase,F3H)在植物花青素合成途径中发挥重要作用,可作为山茶花色遗传育种的候选基因。本研究在前期从Gen Bank数据库中筛选得到白花茶树CsF3H、黄花金花茶CnF3H和红花浙江红山茶CcF3H三个基因。然而,它们的分子遗传与变异信息仍然缺乏,不利于最大正向效应基因的选择利用。本研究系统探讨了CsF3H、CnF3H和CcF3H基因的分子特征、系统进化和蛋白三维结构。结果发现CsF3H、CnF3H和CcF3H三者之间存在高度序列多样性,共含37个核苷酸和9个氨基酸差异,且与CsF3H和CnF3H相比,CcF3H序列发生了更多的变异。系统进化结果表明CsF3H、CnF3H和CcF3H蛋白与猕猴桃AcF3H具有共同祖先,但在后期进化过程中,CcF3H率先与CsF3H和CnF3H二者发生分化。序列比对和保守结构域分析发现CsF3H、CnF3H和CcF3H蛋白均含有一个保守的依赖2-酮戊二酸和二价铁离子的双加氧酶超家族特征区域,该区域内的组氨酸H~(218)和H~(276)以及天冬氨酸D220是Fe~(2+)结合位点,精氨酸R~(286)和丝氨酸S~(288)是2-酮戊二酸的重要结合位点。本研究还发现了3个茶属F3H蛋白的保守结构域中存在一个差异位点,与CsF3H和CnF3H第200位异亮氨酸I200相比,CcF3H对应氨基酸变异为缬氨酸V~(200),Web logo3.4分析揭示V200在植物F3H蛋白中更为保守。空间结构分析显示CcF3H中氨基酸V~(200的替换导致它和赖氨酸K197的分子作用力增强。本研究结果表明CcF3H是更为保守的植物F3H蛋白,缬氨酸V200可能是一个重要的功能位点。上述研究也为山茶属F3H基因提供了新的信息,为今后选择CcF3H基因作为山茶花色育种最适候选基因提供了理论支撑。
[Abstract]:Flavanone 3-hydroxylase gene (F3H) plays an important role in plant anthocyanin synthesis pathway and can be used as a candidate gene for floral color genetics and breeding of Camellia oleifera. In this study, three genes of white flower tea CsF3H, yellow flower gold tea CnF3H and safflower Zhejiang Hongshan tea CcF3H were screened from Gen Bank database. However, their molecular genetic and variation information is still lacking, which is not conducive to the selection and utilization of the maximum positive effect gene. In this study, the molecular characteristics, systematic evolution and protein three-dimensional structure of CsF3H,CnF3H and CcF3H genes were systematically discussed. The results showed that there was a high degree of sequence diversity between CsF3H,CnF3H and CcF3H, which contained 37 nucleotides and 9 amino acids, and CcF3H sequences varied more than CsF3H and CnF3H. The results of systematic evolution showed that CsF3H,CnF3H and CcF3H proteins had a common ancestor with kiwifruit AcF3H, but in the later evolution process, CcF3H first differentiated from CsF3H and CnF3H. Sequence alignment and conserved domain analysis showed that both CsF3H,CnF3H and CcF3H proteins contained a conserved dioxygenase superfamily region dependent on 2-ketoglutaric acid and divalent iron ions. Histidine H218 and H276 and aspartic acid D220 were Fe~ (2) binding sites, arginine R286 and serine S288were important binding sites of 2-ketoglutaric acid. Compared with CsF3H and CnF3H 200th isoleucine I200, the variation of CcF3H corresponding amino acid to valine V200 revealed that V200 was more conserved in plant F3H protein. Spatial structure analysis showed that the substitution of amino acid V200 in CcF3H resulted in the enhancement of the molecular force between V200 and lysine K197. The results showed that CcF3H was a more conservative plant F3 H protein and valine V200 might be an important functional site. These studies also provide new information for Camellia F3 H gene and provide theoretical support for the selection of CcF3H gene as the most suitable candidate gene for Camellia flower color breeding in the future.
【作者单位】： 乐山师范学院生命科学学院;
【基金】：乐山师范学院科研项目(Z1201)资助
【分类号】：Q943.2
本文编号：2519168
[Abstract]:Flavanone 3-hydroxylase gene (F3H) plays an important role in plant anthocyanin synthesis pathway and can be used as a candidate gene for floral color genetics and breeding of Camellia oleifera. In this study, three genes of white flower tea CsF3H, yellow flower gold tea CnF3H and safflower Zhejiang Hongshan tea CcF3H were screened from Gen Bank database. However, their molecular genetic and variation information is still lacking, which is not conducive to the selection and utilization of the maximum positive effect gene. In this study, the molecular characteristics, systematic evolution and protein three-dimensional structure of CsF3H,CnF3H and CcF3H genes were systematically discussed. The results showed that there was a high degree of sequence diversity between CsF3H,CnF3H and CcF3H, which contained 37 nucleotides and 9 amino acids, and CcF3H sequences varied more than CsF3H and CnF3H. The results of systematic evolution showed that CsF3H,CnF3H and CcF3H proteins had a common ancestor with kiwifruit AcF3H, but in the later evolution process, CcF3H first differentiated from CsF3H and CnF3H. Sequence alignment and conserved domain analysis showed that both CsF3H,CnF3H and CcF3H proteins contained a conserved dioxygenase superfamily region dependent on 2-ketoglutaric acid and divalent iron ions. Histidine H218 and H276 and aspartic acid D220 were Fe~ (2) binding sites, arginine R286 and serine S288were important binding sites of 2-ketoglutaric acid. Compared with CsF3H and CnF3H 200th isoleucine I200, the variation of CcF3H corresponding amino acid to valine V200 revealed that V200 was more conserved in plant F3H protein. Spatial structure analysis showed that the substitution of amino acid V200 in CcF3H resulted in the enhancement of the molecular force between V200 and lysine K197. The results showed that CcF3H was a more conservative plant F3 H protein and valine V200 might be an important functional site. These studies also provide new information for Camellia F3 H gene and provide theoretical support for the selection of CcF3H gene as the most suitable candidate gene for Camellia flower color breeding in the future.
【作者单位】： 乐山师范学院生命科学学院;
【基金】：乐山师范学院科研项目(Z1201)资助
【分类号】：Q943.2
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