作物学报2017，Vol.43Issue(5)：640-647,8.DOI:10.3724/SP.J.1006.2017.00640
甘蓝型油菜烷羟化酶基因MAH1的克隆与表达分析
Cloning and Expression Analysis of Alkane Hydroxylase Gene MAH1 from Brassica napus
摘要
Abstract
In wax biosynthesis, the mid-chain alkane hydroxylase (MAH) catalyzes the hydroxylation reaction from alkanes tosecondary alcohols and further to corresponding ketones. In this study, using Arabidopsis P450-dependent enzyme CYP96A15/ MAH1 gene as a probe, the full-length coding sequences of two Brassica napus MAH1 genes, BnMAH 1-1 (GenBank accession number KT795344) and BnMAH1-2 (GenBank accession number KT795345), were isolated by in silico cloning and RT-PCR. The ORF lengths of BnMAH1-1 and BnMAH1-2 were 1491 bp, and no intron was included. BnMAH1-1 and BnMAH1-2 shared 92.4% and 90.9% of sequence identity at nucleotide and amino acid level, respectively. The predicted BnMAH1-1 and BnMAH1-2 protein contained 496 amino acid residues, with typical P450 protein family conserved domains P415xR417x, K helix (E359xxR362), C-terminal hemopexin-like domain (F436xxGxRxCxG445) and oxygen binding zone (A/G)G309x(D/E)T312(T/S). NCBI BlastN,multiple alignment of amino acid sequence, and phylogenetic analysis showed that they were most homologous to A. thaliana CYP96A15/MAH1. Real-time quantitative PCR showed that BnMAH1-1 and BnMAH1-2 were mainly expressed in stem, leaf, flower, and silique. The highest expression level was found in leaf and the lowest was in root, which was consistent with the wax deposition on aerial part of plant. The expression of BnMAH1-1 and BnMAH1-2 was barely detected in stem and leaf of wax deficient cultivar, suggesting that the reduced wax deposition is due to the down-regulation of MAH1 transcription. The expression of BnMAH1-1 and BnMAH1-2 was significantly induced by exogenous application of SA, MeJA, ACC, and ABA and exposure to NaCl and drought, among which BnMAH1-1 may play a major role in response to water stress.关键词
甘蓝型油菜/角质层蜡质/烷羟化酶Key words
Brassica nupas/Cuticular wax/Alkane hydroxylase引用本文复制引用
徐熠,彭阳,李帅,赵秋棱,张双娟,李加纳,倪郁..甘蓝型油菜烷羟化酶基因MAH1的克隆与表达分析[J].作物学报,2017,43(5):640-647,8.基金项目
本研究由重庆市基础与前沿研究计划项目(cstc2016jcyjA0170)和中央高校基本科研业务费专项资金(XDJK2014B037)资助. This study was supported by the Chongqing Basic and Advanced Research Project (cstc2016jcyjA0170) and the Fundamental Research Funds for the Central Universities (XDJK2014B037). （cstc2016jcyjA0170）
