苹果SUPERMAN类蛋白家族的基因克隆与生物信息学分析  

Gene Cloning and Bioinformatics Analysis of SUPERMAN Family Proteins in Apple(Malus domestic Borkh.)

作　　者：赵旭勉[1] 任雪芹[1] 张文[1] 李皓[1] 朱元娣[1] 

机构地区：[1]中国农业大学农学与生物技术学院果树系,北京100193

出　　处：《农业生物技术学报》2012年第7期782-790,共9页Journal of Agricultural Biotechnology

基　　金：国家公益性行业(农业)专项经费(201003021)

摘　　要：SUPERMAN 类基因是植物生长发育中的重要转录因子。为了揭示苹果 SUPERMAN 类蛋白家族的生理基因功能,本研究以富士苹果(Malus domestica Borkh.cv. Fuji)为试材,利用 RT-PCR 方法和锚定PCR 方法克隆了 MdLIFa(GenBank 登录号 HM370493)和启动子序列,利用特异引物 PCR 方法克隆了苹果基因组中其余 11 个 SUPERMAN 类基因。12 个基因命名为 MdLIFa/MdLIFb (GenBank 登录号:HQ260429)、MdSUP1a (HQ418477)/MdSUP1b (HQ418478)、MdSUP4 (HQ418475)/MdSUP12 (HQ418476)、MdSUP5a (HQ418469)/MdSUP13b (HQ418470)、MdSUP9a (HQ418473)/MdSUP9b (HQ418474) 和 MdSUP5b(HQ418471)/MdSUP13a(HQ418472),分别位于 11/3、1/1、4/12、5/13、9/ 未知和 5/13 号染色体,每对基因间的同源性均在86%以上。通过基因枪法轰击洋葱(Allium cepa)表皮细胞,基因瞬时表达证明 MdLIFa 定位于细胞核内。MdLIFa 基因启动子序列中含有多个参与花粉发育、根特异性表达、与光诱导和 Dof 单锌指基因家族作用的调控元件等。系统进化分析表明,MdLIFa/b、MdSUP9a/9b 和 MdSUP5a/13b、以及MdSUP1a/b、MdSUP5b/MdSUP13a和 MdSUP4/12 分别聚类为一组,参与植株的形态建成和花器官发育。苹果 12 个 SUPERMAN 类基因成对高度同源,具有生物学功能的多样性,为深入的基因鉴定和遗传转化提供有用信息。SUPERMAN family genes are important transcription factors in the plant growth and development. In order to understand physiological functions of SUPERMAN family genes in apple （Malus domestica Borkh.）, the MdLIFa （GenBank accession No. HM370493） and its promoter were cloned using RT-PCR and anchored PCR from apple cultivar Fuji （M. domestico~ cv. Fuji）. The 11 genes of SUPERMAN family in apple were cloned by PCR amplification using gene-specific primers. All 12 genes were designated as MdLIFaJMdLIFb （GenBank accession No. HQ260429）, MdSUPla （HQ418477）/MdSUP1b （HQ418478）, MdSUP4（HQ418475） /MdSUP12 （HQ418476）, MdSUP5a （HQ418469）/MdSUP13b （HQ418470）, MdSUP9a（HQ418473）/MdSUPgb （HQ418474）, and MdSUP5b （HQ418471）/MdSUP13a （HQ418472）, located on chromosomes 11/3, 1/1, 4/12, 5/13, 9/unknown, and 5/13, respectively. The similarity of deduce amino acid sequences of each pair of genes was higher than 86%. The subcellular localization of MdLIFa was transient expressed in the cell nucleus by the particle bombardment into onion （Allium cepa） epidermal cells. Cis-acting elements, such as POLLEN1 LELAT52 （AGAAA）, GTGANTG10 （GTGA）, ROOTMOTIFTAPOX1 （ATATT）, DOFCOREZM （AAAG）, and GT 1 CONSENSUS （GRWAAW）, were found in the upstream of MdLIFa. Phylogenetic analysis showed that MdLIFa/b were clustered in one group, together with MdSUP9a/9b and MdSUP5a/13b while MdSUPla/b, MdSUP5b/MdSUP13, and MdSUP4/12 in another group. These genes were putatively involved in plant morphogenesis and floral organogenesis. The SUPERMAN family genes in apple are highly homologous in pair with diverse biological functions, which provide useful information for further gene characterization and genetic transformation.

关 键 词：苹果 SUPERMAN类基因 转录因子 生物信息学 

分 类 号：S661.1[农业科学—果树学]