石蒜LrCMO基因的克隆及表达分析  

Cloning and Expression Analysis of CMO Gene from Lycoris radiata

作　　者：徐晟[1] 蒋明敏[1] 江宜龙 夏冰[1] 汪仁[1] 

机构地区：[1]江苏省中国科学院植物研究所,南京210014

出　　处：《分子植物育种》2017年第4期1240-1248,共9页Molecular Plant Breeding

基　　金：国家自然科学基金资助项目(31301798);江苏省第四期"333"工程培养资金资助项目(BRA2015432);江苏省省属公益院所科研条件与能力建设项目(BM2015019)共同资助

摘　　要：以石蒜(Lycoris radiata)为试材,采用同源克隆和RACE的试验方法,克隆得到LrCMO基因全长cDNA,并对其进行了基因表达分析。结果表明:LrCMO基因全长1 472 bp,其中开放阅读框(ORF)为1 281 bp,编码427个氨基酸残基,预测编码蛋白质的分子量为47.92 kD,理论等电点为5.72;LrCMO是一个稳定的疏水蛋白,不具有跨膜结构,含有叶绿体导肽;LrCMO基因编码的氨基酸与植物其他CMO蛋白具有较高的一致性,且与海枣PdCMO、香蕉MaCMO及油棕EgCMO亲缘关系最高,聚为一类。实时荧光定量PCR分析表明,LrCMO在根、鳞茎和叶片中有表达,且在鳞茎中的表达量最高;LrCMO受聚乙二醇(PEG)处理的诱导表达,其基因相对表达量在处理后12 h达到最高。随着处理时间的延长,LrCMO基因相对表达量逐渐下调至对照水平。The full length cDNA sequence of CMO was obtained from Lycoris radiata based on homology cloning and RACE method and its expression patterns were also analyzed.The results showed that the full-length cDNA of LrCMO was 1 472 bp,containing a 1 281 bp open reading frame（ORF） which encoded 427 amino acids with a predicted molecular weight of 47.92 kD and p I 5.72.LrCMO was a stable hydrophobic protein,and had a chloroplast transit peptide and no tranmembrane structure.Multiple sequence alignment and phylogenetic tree analysis showed that the deduced protein LrCMO shared higher identity with CMO from other plants,and belongs to the same branch with Phoenix dactylifera Pd CMO,Camellia sinensis CsCMO and Elaeis guineensis EgCMO.Subsequently,quantitative real-time PCR analysis indicated that LrCMO was expressed in leaves,bulbs,and roots with the highest expression level in bulbs.LrCMO transcript levels were significantly induced by 20% polyethylene glycol（PEG） treatment,and peaked in 12 h treatment.

关 键 词：石蒜 LrCMO基因克隆 甜菜碱 序列分析 

分 类 号：Q943.2[生物学—植物学] S682.29[农业科学—观赏园艺]