大白菜番茄红素β-环化酶基因 BrLCYB的鉴定与功能分析  

Identification and functional analysis of the BrLCYB gene of lycopene β -cyclase from Chinese cabbage

作　　者：李必元[1] 岳智臣[1] 赵彦婷[1] 雷娟利[1] 胡齐赞[1] 陶鹏[1] LI Biyuan;YUE Zhichen;ZHAO Yanting;LEI Juanli;HU Qizan;TAO Peng(Institute of Vegetables,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China)

机构地区：[1]浙江省农业科学院蔬菜研究所,浙江杭州310021

出　　处：《浙江农业学报》2023年第9期2090-2096,共7页Acta Agriculturae Zhejiangensis

基　　金：浙江省农业新品种选育重大科技专项(2021C02065-5-2);浙江省自然科学基金(LY21C150006);浙江省农业重大技术协同推广计划(2021XTTGSC01-2)。

摘　　要：本研究从大白菜中克隆了唯一的番茄红素β-环化酶基因(BrLCYB),编码序列长度为1482 bp,BrLCYB只在5′UTR所对应的基因序列中存在一个内含子,导致其转录后形成2种类型的转录本(BrLCYB-1和BrLCYB-2),其中BrLCYB-1类型占98.26%。BrLCYB广泛表达在多种器官中和整个胚胎发育阶段,其中在花和成熟胚胎中有着较高的表达。系统进化树显示,高等植物的LCYB起源于蓝藻,LCYB基因在进化过程中出现了内含子获得和内含子丢失事件,最终在高等植物的编码序列中丢失全部内含子。BrLCYB蛋白包含番茄红素β-环化酶结构域,为了进一步验证BrLCYB的功能,构建了pET-BrLCYB原核表达载体。pACCRT-EIB和pET-BrLCYB共转化到大肠埃希菌中,可使得大肠埃希菌富集橙黄色的β-胡萝卜素,证明大白菜BrLCYB具有番茄红素β-环化酶的催化功能。In this study,the only lycopeneβ-cyclase was cloned from Chinese cabbage(BrLCYB)and had 1482 bp coding sequence.BrLCYB contains the only one intron in its gene sequence,corresponding to 5′UTR.The intron resulted in BrLCYB producing two types of transcripts(BrLCYB-1 and BrLCYB-2)after transcription.BrLCYB-1 accounted for 98.26%of the total BrLCYB transcripts.BrLCYB was widely expressed in a variety of organs and the whole embryonic development stage,and has a higher expression in flowers and mature embryos.The phylogenetic tree of LCYBs showed that the LCYBs of higher plants originated from cyanobacteria.During the evolution of LCYB genes,intron gain and intron loss,occurred successively in LCYB s,and finally no intron existed in coding sequences of LCYB s in higher plants.BrLCYB contained a lycopeneβ-cyclase domain.In order to further verify the function of BrLCYB,the pET-BrLCYB prokaryotic expression vector was constructed.The co-transformation of pACCRT-EIB and pET-BrLCYB into E.coli can make E.coli enriching orange-yellowβ-carotene,proving that BrLCYB of Chinese cabbage had the function of lycopeneβ-cyclase.

关 键 词：大白菜 番茄红素Β-环化酶 基因 功能分析 

分 类 号：S634.1[农业科学—蔬菜学]