唐菖蒲丙二烯氧化物环化酶基因GhAOC的克隆与表达分析  

Cloning and expression analysis of allene oxide cyclase gene GhAOC from Gladiolus hybridus

作　　者：连青龙[1,2] 李晓昕[2] 钟雄辉[2] 尹义蕾[1] 义鸣放[2] 

机构地区：[1]农业部规划设计研究院设施农业研究所,北京100125 [2]中国农业大学农学与生物技术学院,北京100193

出　　处：《中国农业大学学报》2012年第5期46-53,共8页Journal of China Agricultural University

基　　金：公益性行业(农业)科研专项(200903020;200903009)

摘　　要：克隆唐菖蒲AOC基因的全长cDNA序列,并分析该基因的表达模式及其对茉莉酸(Jasmonic acid,JA)生物合成的影响。以唐菖蒲栽培品种‘Rose Supreme’的球茎为试材,利用RT-PCR和RACE技术克隆AOC基因的全长cDNA序列;利用基因枪方法进行GhAOC基因的亚细胞定位分析;运用实时荧光定量PCR技术分析GhAOC基因的表达模式。克隆了1个全长为898bp的茉莉酸生物合成关键酶丙二烯氧化物环化酶(allene oxide cyclase,AOC)基因的cDNA序列,命名为GhAOC。该序列含有1个735bp的开放阅读框(ORF),编码244个氨基酸,推导的蛋白质分子量为26.52ku。亚细胞定位分析表明GhAOC是叶绿体蛋白。实时荧光定量RT-PCR表达分析显示,该基因在唐菖蒲叶、花、根、匍匐茎、新球茎和籽球上都表达,其中在新球茎和籽球中表达量最高;经过0.1～0.5mmol/L的MJ(methyl jasmonate,茉莉酸甲酯)处理后,逐渐提高了GhAOC基因在球茎中的表达量和内源MJ含量。GhAOC基因的表达促进了唐菖蒲茉莉酸的生物合成。To clone the full-length cDNA of GhAOC and analyze the expression patterns of GhAOC and the effect on the biosynthesis of jasmonic acid.The full length cDNA of GhAOC was cloned in Gladiolus hybridus’Rose Supreme’corms by RT-PCR and RACE.The technology of gene gun bombardment was used to analyze the sub-cellular localization of GhAOC.The real time RT-PCR was used to analyze the expression pattern of GhAOC.A full-length cDNA named GhAOC encoding a key enzyme of the biosynthesis of jasmonic acid was cloned in Gladiolus.The open reading frame encompassed 735 bp encoding a polypeptide of 244 amino acids with calculated protein molecular mass of 26.52 ku.The sub-cellular localization analysis indicated that GhAOC was a chloroplast protein.Real time RT-PCR analysis showed that GhAOC gene was expressed in leaf,flower,root,stolon,corm and cormel,and the relatively high expression level of GhAOC was observed in corm and cormel.Meanwhile,the expression level and the endogenous MJ content in corms steadily increased under MJ treatment with a raising concentration gradients from 0.1 mmol/L to 0.5 mmol/L.Based on the results of our pilot study,the expression of GhAOC promoted the the biosynthesis of jasmonic acid in Gladiolus hybridus.

关 键 词：唐菖蒲 AOC基因 JA 克隆 表达 

分 类 号：S682.2[农业科学—观赏园艺]