香椿查尔酮异构酶基因的克隆及在幼苗中的表达分析  

Cloning and Expression Analysis of Chalcone Isomerase Gene in Toona sinensis Seedlings

作　　者：隋娟娟 屈长青 杨京霞 舒生 董新月 姜景民[2] 刘军[2] Sui Juanjuan;Qu Changqing;Yang Jingxia;Shu Sheng;Dong Xinyue;Jiang Jingmin;Liu Jun(Engineering Technology Research Center of Anti-aging Chinese Herbal Medicine,Fuyang Normal University,Fuyang,236037;Key Laboratory of Tree Subtropical Breeding and Cultivation National Forestry and Grassland Administration,Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou,311400)

机构地区：[1]阜阳师范大学,抗衰老中草药安徽省工程技术研究中心,阜阳236037 [2]中国林业科学研究院亚热带林业研究所,国家林业和草原局亚热带林木培育重点实验室,杭州311400

出　　处：《分子植物育种》2021年第14期4649-4656,共8页Molecular Plant Breeding

基　　金：国家林业和草原局亚热带林木培育重点实验室开放基金(RISF2019PY02);安徽高校自然科学研究重点项目(KJ2019A0516,KJ2020A0520);安徽省科技重大专项(202003a06020020)共同资助。

摘　　要：查尔酮异构酶是调控植物花青素生成的关键酶,本研究利用PCR扩增技术从香椿叶片中克隆得到香椿查尔酮异构酶基因TsCHI。序列分析发现TsCHI基因开放阅读框全长717 bp,共编码238个氨基酸,属于亲水性、酸性蛋白。系统进化分析表明TsCHI蛋白属于TypeⅠ型CHI蛋白,与无患子目的槭树(Acer×freemanii)、橄榄(Canarium album)、龙眼(Dimocarpus longan)、柑橘(Citrus sinensis)的亲缘关系较近。实时荧光定量PCR结果表明,TsCHI在香椿幼苗茎中的表达量显著高于根与叶中的表达量,叶比根中的表达量高,但差异不显著;38℃高温胁迫、200 g/L PEG-6000溶液干旱胁迫、200 mmol/L NaCl溶液盐胁迫条件下,TsCHI基因的表达量均高于对照,且随着处理时间的延长,表达呈上升趋势。研究结果为香椿幼苗的开发利用和通过基因工程手段提高幼苗中黄酮类物质的含量提供了理论参考。Chalcone isomerase is one of the key enzymes in the regulation of anthocyanin production.In this study,the chalcone isomerase gene(TsCHI)was cloned from Toona sinensis leaves by PCR to understand its bioinformatics and expression character in Toona sinensis seedlings.Sequence analysis showed the open reading frame of TsCHI gene contained 717 bp nucleotides,encoding 238 amino acids,and the TsCHI protein belonged to hydrophilic acidic protein.Phylogenetic analysis showed that TsCHI belonged to TypeⅠCHI protein and had the closest relationship with Acer×freemanii,Canarium album,Dimocarpus longan and Citrus sinensis,which all belonged to Sapindales.The results of real-time PCR showed the TsCHI gene expression level in the stems was significantly higher than that in the roots and leaves;The expression in leaves was higher than that in roots,but the difference was not significant;Under high temperature stress with 38℃,drought stress with 200 g/L PEG-6000 solution and salt stress with 200 mmol/L NaCl solution,the expression of TsCHI gene was higher than that of the control,and the expression showed an upward trend with the extension of processing time.The results of this study provide theoretical reference for the Toona sinensis seedlings utilization and the theoretical reference for improvement of flavonoids content in the Toona sinensis seedlings by means of molecular breeding.

关 键 词：香椿(Toona sinensis) 查尔酮异构酶 花青素 表达分析 

分 类 号：S644.4[农业科学—蔬菜学]