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为研究兰花成花转变及花发育的调控机理,利用反转录RT-PCR和RACE的方法,从蕙兰萼片中克隆出一个APETALA1/FRUITFULL-like(AP1/FUL-like)基因,命名为CfAP11,GenBank登录号为JQ031272.1。该基因编码的氨基酸序列与MADS-box蛋白家族中类AP1/FUL亚家族中球花石斛FRUITFULL-like具有较高的同源性(84%),系统进化分析表明该蛋白的氨基酸序列与AP1/FUL转录因子亚家族中的蛋白聚为一类。生物信息学分析推测表明,该基因编码的蛋白具有MADS保守域和相对保守的K区,二级结构中-螺旋所占比例较高(58.97%),三级结构与月季、水稻和水仙非常相似。相对荧光定量PCR分析结果表明:CfAP11在根中表达痕量,生殖期比营养期叶片中表达量低、盛花期比花蕾期花葶中表达量高,由此推测,CfAP11可能与蕙兰的成花诱导、花发育有关;并发现CfAP11在盛花期花葶和子房中表达量远高于其他组织,表明其可能以某种机制参与果实的形成过程。
In order to study the regulation mechanism of flowering and flower development of orchids, an APETALA1 / FRUITFULL-like (AP1 / FUL-like) gene was cloned from Cymbidium septenbagh by reverse transcription RT-PCR and RACE and named as CfAP11 GenBank accession number is JQ031272.1. The amino acid sequence of this gene has high homology (84%) with FRUITFULL-like of Dendrobium candidum in the AP1 / FUL subfamily of MADS-box protein family. Phylogenetic analysis showed that the amino acid sequence of this gene is homologous with AP1 / The proteins in the FUL transcription factor subfamily are clustered together. Bioinformatics analysis suggested that the protein encoded by this gene possessed the MADS conserved domain and the relatively conserved K domain. The secondary structure contained a high proportion of helices (58.97%), and the tertiary structure was very similar to that of rose, rice and narcissus . Relative fluorescence quantitative PCR analysis showed that: CfAP11 expression in the roots trace, the reproductive phase than the vegetative leaf expression is low, full-flowering bud flower buds in the high expression level, suggesting that CfAP11 and Cymbidium into The results showed that the expression level of CfAP11 was significantly higher than that of other tissues in flowering stage and ovary, indicating that it may participate in the formation of fruit with some mechanism.