甘蓝型油菜谷胱甘肽过氧化物酶基因克隆和非生物胁迫下的表达  

Glutathione peroxidase gene BnGPX1 cloning from rapeseed and expression under abiotic stress

作　　者：胡茂龙[1] 龙卫华[1] 高建芹[1] 陈新军[1] 张洁夫[1] 陈松[1] 戚存扣[1] 浦惠明[1] 

机构地区：[1]江苏省农业科学院经济作物研究所,国家油料作物改良中心南京分中心,江苏南京210014

出　　处：《中国油料作物学报》2011年第4期331-337,共7页Chinese Journal of Oil Crop Sciences

基　　金：国家科技支撑计划(2009BADA8B01);江苏省科技支撑计划(BE2008369)

摘　　要：利用RT-PCR(reverse transcription PCR)技术从甘蓝型油菜宁油16号(Brassica napus L.)中克隆了一个非生物胁迫下细胞抵御活性氧(reactive oxygen species,ROS)伤害的关键酶GPX(谷胱甘肽过氧化物酶)基因,命名为BnGPX1(GenBank登录号:HM130680)。BnGPX1的开放阅读框长度为711bp,推测编码蛋白含有236个氨基酸,分子量为26.10kD,等电点为9.37。BnGPX1酶具有GPX特有的3个结构域及半胱氨酸残基。通过PCR方法克隆得到BnGPX1基因组序列(GenBank登录号:HM130681)。该序列与拟南芥AtGPX1基因相似,由6个外显子和5个内含子组成,内含子的剪切位点符合真核生物GT-AG规则。半定量RT-PCR发现BnGPX1在油菜茎、叶、角果和花中表达量高,根中表达量较低。在盐、干旱和高温胁迫下BnGPX1表达量升高,出现峰值的时间不同。该基因对低温胁迫没有响应。A glutathione peroxidases gene,designated as BnGPX1（GenBank assession number HM130680）,was a key enzyme protecting plants against oxidative damage generated by reactive oxygen species（ROS） under abiotic stresses.It was cloned from rapeseed cultivar Ningyou 16（Brassica napus L.） by RT-PCR（reverse transcription PCR）.The open reading frame（ORF） of BnGPX1 was 711 bp,encoding 236 amino acids with isoelectric point（pI） of 9.37 and molecular mass of 26.10 kD.BnGPX1 had 3 characteristic domains of plant GPXs and 3 conserved cysteine residues.Its genomic DNA（gDNA） fragment（GenBank assession number HM130681） was isolated by PCR.The sequence of gDNA demonstrated 6 exons separated by 5 introns.All introns were spliced following the consensus sequence with GT and AG at 5＇ and 3＇ ends respectively.BnGPX1 construction was similar to the Arabidopsis AtGPX1 gene.Expression patterns of BnGPX1 in rapeseed tissues under different abiotic stresses were obtained by semi-quantitative RT-PCR.Results showed that BnGPX1 was constitutively and ubiquitously expressed at high levels in stems,leaves,silliques and flowers,but at lower level in roots.The expression was increased under salt,drought and high temperature condition.It did not respond to cold stress.

关 键 词：甘蓝型油菜 谷胱甘肽过氧化物酶 表达 非生物胁迫 

分 类 号：Q781[生物学—分子生物学] S565.4[农业科学—作物学]