杉木BR生物合成酶ClDET2基因的克隆及表达分析  

Cloning and Expression Analysis of the BR Biosynthetic Enzyme Gene ClDET2 in Chinese Fir

作　　者：理挪 林思祖[2] 张健 张家君 吕蒙 陈宇[2] Li Nuo;Lin Sizu;Zhang Jian;Zhang Jiajun;LU Meng;Chen Yu(College of Forestry,Fujian Agriculture and Forestry University,Fuzhou,350002;State Forestry Administration Engineering Research Center of Chinese Fir,Fuzhou,350002)

机构地区：[1]福建农林大学林学院,福州350002 [2]国家林业局杉木工程技术研究中心,福州350002

出　　处：《分子植物育种》2019年第24期8058-8064,共7页Molecular Plant Breeding

基　　金：国家林业局杉木工程技术研究中心平台建设(ptjh130002)资助

摘　　要：类固醇5α-还原酶(DET2)是BR生物合成的限速酶,是调控油菜素内酯生物合成的关键基因。为了了解油菜素内酯(BR)生物合成途径中限速酶DET2的作用,本研究以杉木低涩籽率家系L为材料,采用RTPCR法从杉木中克隆获得一个转录因子基因并命名为ClDET2。分析发现,该序列包含一个长度为840 bp的CDS基因序列(开放阅读框),编码215个氨基酸,系统进化树分析显示,ClDET2与NCBI数据库中注释的北美云杉DET2基因亲缘关系最近。通过对ClDET2基因在种子发育的4个时期进行qPCR表达检测,发现其表达量从种子发育到成熟呈总体下降的趋势。这可能是ClDET2基因主要在种子发育前期的胚珠发育阶段表达高的原因,而6~7月份杉木种子胚胎发育阶段多涩粒产生,也可能与ClDET2表达量降低有关。以上结果为进一步研究杉木ClDET2基因的功能,以及阐明涩籽产生的分子机理提供了科学依据。Steroids 5α-reductase(DET2)is the rate-limiting enzyme in BR biosynthesis and is a key gene regulating the biosynthesis of brassinolide.This study aimed to understand the role of the rate-limiting enzyme DET2 in the brassinolide(BR)biosynthesis pathway.In this study,we obtained a bp-conserved gene fragment of cedar and a 840 bp CDS gene sequence by RT-PCR with a low sorghum seed rate family of L,and its open reading frame was 648 bp,encoding 215 amino acids.Phylogenetic tree analysis showed that ClDET2 was most closely related to the annotated North American spruce DET2 gene in the NCBI database.Through the qPCR expression of ClDET2 gene in the four stages of seed development,it was found that the expression level decreased from seed development to maturity.This may be due to the fact that the ClDET2 gene is mainly expressed in the early stage of ovule development in the early stage of seed development,and the emergence of multiple grains in the embryonic development stage of Chinese fir from June to July may also be related to the decreased expression of ClDET2.These results provide a scientific basis for further study of the function of the ClDET2 gene of Chinese fir and the molecular mechanism of the production of alfalfa seeds.

关 键 词：杉木(Cunninghamia lanceolata) ClDET2 基因克隆 表达分析 

分 类 号：S791.27[农业科学—林木遗传育种] Q943.2[农业科学—林学]