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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。
The common APD / AP gene AcDEF was cloned by RACE and the expression pattern of AcDEF in various organs of onion was studied by semi-quantitative RT-PCR and real-time fluorescence quantitative PCR. The AcDEF gene of GenBank accession number JX661502 has a length of 1 014 bp and an open reading frame of 675 bp encoding a protein of 224 amino acids. Phylogenetic analysis showed that the AcDEF gene belonged to the AP3 / DEF subfamily of monocot B gene families. Expression pattern analysis showed that AcDEF was expressed specifically in floral organs, with high abundance in petals and stamens, slight expression in flower buds and membranous involucre, and very low expression in carpels. In vegetative organs such as roots , False stems, leaves and bulbs in the expression. During flowering, the accumulation of AcDEF transcripts in various floral organs showed dynamic changes. AcDEF was highly abundant in petals and stamens, but slightly decreased in the petals and stamens of fully open flowers. The expression of AcDEF in membranous involucre and carpel of the completely open flower is very low in the total bud and the carpels. The sequence structure and expression pattern of AcDEF gene are characteristic of the monocot AP3 / DEF gene and belong to the paleoAP3 evolutionary line.