洋葱花发育B类MADS-box基因AcDEF的克隆与表达分析  

Cloning and Expression Analysis of B Class MADS-box Gene AcDEF Associated with Floral Development in Onion

作　　者：徐启江[1] 丁一[1] 黄赫[1] 王勇[2] 崔成日 梁毅[4] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040 [2]东北农业大学园艺学院,哈尔滨150030 [3]哈尔滨长日洋葱研究所,哈尔滨150090 [4]北京市农林科学院蔬菜研究中心,北京100097

出　　处：《园艺学报》2013年第6期1090-1100,共11页Acta Horticulturae Sinica

基　　金：黑龙江省自然科学基金项目(C201016);农业部公益性行业项目(200903018-6-5);哈尔滨市创新人才研究专项(2013RFLYN015)

摘　　要：以紫皮洋葱常规品种‘RUPI’为试材,通过 RACE 方法克隆了 AP3/DEF 同源基因 AcDEF,并用半定量 RT-PCR 和实时荧光定量 PCR 研究 AcDEF 在洋葱各类器官中的表达模式。GenBank 登录号为JX661502 的 AcDEF 基因长 1 014 bp,开放阅读框长度为 675 bp,编码 224 个氨基酸。系统发育分析表明,AcDEF 基因属于单子叶 B 功能基因家族的 AP3/DEF 亚家族。表达模式分析显示,AcDEF 在花器官中特异表达,其中在花瓣、雄蕊内高丰度表达,在花葶和膜状总苞中微量表达,在心皮中表达水平极低,在无性营养器官如根、假茎、叶片和鳞茎中无表达。在开花过程中,各花器官中 AcDEF 转录物的积累呈动态变化;在花瓣和雄蕊发育过程中 AcDEF 均高丰度表达,但在完全开放花的花瓣和雄蕊中表达量略有降低;在花葶、膜状总苞和心皮中表达量递减,在完全开放花的膜状总苞和心皮中 AcDEF 的表达量很低。AcDEF 基因的序列结构和表达模式具有单子叶物种 AP3/DEF 基因的特征,属于 paleoAP3 进化系。We isolated the AP3/DEF-homologue from the onion variety ＇RUPI＇ using RACE （rapid amplification of cDNA ends）, and characterized its expression patterns in vegetative and floral organs using semi-quantitative RT-PCR and quantitative real-time PCR. Phylogenetic analysis indicates that our onion gene, denoted AcDEF, belongs to the AP3/DEF subfamily of the B-function gene family and has sequence characteristics similar to other monocot paleoAP3 genes. The cDNA sequence is 1 014 bp long, includes both 5＇ and 3＇ untranslated regions, a poly （A） tail, and an open reading frame encoding a protein with a predicted length of 224 amino acids. Expression analyses indicate that AcDEF is specifically expressed in flower with no detectable signal in vegetative organs, such as roots, cauloids, leaves and bulbs In addition, expression is tissue-specific in floral organs, with the highest level in the petals andstamens, moderate level in scapes and membranous sheath. AcDEF is expressed at very low levels in the carpels. Accumulation of AcDEF transcripts is dynamically changed and associated with the flower-bud formation and development. Expression of AcDEF is strongly detected during the initiation and early development of petals and stamens, but expression levels in these organs are somewhat reduced at later developmental stages. There also appears to be a gradual and obvious decrease in AcDEF expression during the different developmental stages of scapes, membranous sheath and carpels, such that it is almost absent from the membranous sheath and carpels in full-opening flowers.

关 键 词：洋葱 花发育 B-功能MADS—box基因 AcDEF基因 

分 类 号：S633.2[农业科学—蔬菜学]