灰楸CfPPR1基因的克隆及表达分析  

Cloning and Expression Analysis of CfPPR1 Gene of Catalpa fargesii

作　　者：赵林姣 杨英英 付鹏跃 张玉 费越 王楠[2] 张大才 王军辉[2] ZHAO Lin-jiao;YANG Ying-ying;FU Peng-yue;ZHANG Yu;FEI Yue;WANG Nan;ZHANG Da-cai;WANG Jun-hui(Faculty of Forestry,Southwest Forestry University,Kunming 650224;State Key Laboratory of Tree Genetics and Breeding,Research Institute of Forestry,Chinese Academy of Forestry,Key Laboratory of Tree Breeding and Cultivation of State Forestry and Grassland Administration,Beijing 100091;Biotechnology Research Center of China Three Gorges University,Yichang 443002;State Key Laboratory of Tree Genetics and Breeding,Northeast Forestry University,Harbin 150040)

机构地区：[1]西南林业大学林学院,昆明650224 [2]林木遗传育种国家重点实验室,中国林业科学研究院林业研究所,国家林业和草原局林木培育重点实验室,北京100091 [3]三峡大学生物技术研究中心,宜昌443002 [4]东北林业大学林木遗传育种国家重点实验室,哈尔滨150040

出　　处：《温带林业研究》2022年第1期21-29,共9页Journal of Temperate Forestry Research

基　　金：樟楠木等珍贵树种彩叶新品种选育与产业化关键技术(2021SY02)。

摘　　要：【目的】挖掘调控‘麦缘锦楸’黄色边缘形成的关键基因,明确该候选基因的保守结构域、进化关系及表达特征,为下一步揭示该基因的分子生物学功能奠定基础。【方法】基于前期RNA-seq数据,筛选并克隆了与叶色相关的三角状五肽重复(pentatricopeptide repeat,CfPPR1)基因全长;利用ExPASy、InterProScan和CBS等在线工具对CfPPR1进行等电点、蛋白质结构及不同物种同源序列的进化关系和保守结构域分析;比较了CfPPR1基因在不同光照强度下的表达模式。【结果】该CDS序列全长为1902 bp,编码633个氨基酸,推测蛋白相对分子质量为71.31 kD,理论等电点(pI)为7.26;二级结构分析发现,CfPPR1序列编码的蛋白质空间结构是相对稳定的。系统进化树结果显示,CfPPR1不仅与黄化风铃木、芝麻、丹参等菊分支植物聚为一支,还与非菊分支的苹果、桃和橡胶树等具有较高的同源性,说明CfPPR1具有较高的保守性。对CfPPR1基因的表达模式分析发现,CfPPR1基因表达的上调很可能参与调控了‘麦缘锦楸’黄色边缘的形成。【结论】首次克隆得到了与叶色相关的CfPPR1基因,该基因的获得为进一步后期转基因验证CfPPR1基因功能提供有价值的参考。【Objective】The key genes that regulate the formation of the yellow edge of‘Maiyuanjinqiu’were excavated,and the conserved domain,evolutionary relationship and expression characteristics of the candidate gene were identified,which laid the foundation for the next step to reveal the molecular biological function of the gene.【Method】Based on the previous RNA-seq data,the full-length pentatricopeptide repeat(CfPPR1)gene related to leaf color was screened and cloned.The isoelectric point,protein structure and analysis of CfPPR1 were performed using online tools such as ExPASy,InterProScan and CBS.Evolutionary relationship and conserved domain analysis of homologous sequences in different species;the expression patterns of CfPPR1 gene under different light intensities were compared.【Result】The full length of the CDS sequence was 1902 bp,encoding 633 amino acids.The predicted relative molecular mass of the protein was 71.31 kD,and the theoretical isoelectric point(pI)was 7.26.Secondary structure analysis showed that the spatial structure of the protein encoded by the CfPPR1 sequence was relatively stable.The results of the phylogenetic tree showed that CfPPR1 not only clustered with chrysanthemum branch plants such as Handroanthus chrysanthus,Sesamum indicum,and Salvia miltiorrhiza,but also had high homology with non-chrysanthemum branched Malus domestica,Amygdalus persica,and Hevea brasiliensis,indicating that CfPPR1 was highly conserved.Analysis of the expression pattern of the CfPPR1 gene found that the up-regulation of CfPPR1 gene expression is likely to be involved in the regulation of the formation of the yellow edge of‘Maiyuanjinqiu’.【Conclusion】The CfPPR1 gene related to leaf color was cloned for the first time,and the acquisition of this gene provides a valuable reference for further transgenic verification of CfPPR1 gene function in the later stage.

关 键 词：‘麦缘锦楸’ CfPPR1 克隆 基因表达 

分 类 号：S722.99[农业科学—林木遗传育种]