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以黄金树的花芽为材料,采用同源基因克隆技术,获得黄金树花器官特征决定的AG同源基因,将其命名为CaspAG,其开放阅读框(ORF)为738 bp,编码245个氨基酸。分子系统发生和蛋白序列比对分析表明:CaspAG是拟南芥的AG同源蛋白,被归为eu AG进化分支,其MADS区有57个氨基酸,I区有32个氨基酸,K区有83个氨基酸,C区有55个氨基酸,其中C末端的转录激活区含有两个保守的基序:AGI和AGII基序。半定量RT-PCR分析表明,在花发育过程中,CaspAG基因仅在雄蕊和雌蕊中表达,而在茎、叶片、萼片和花瓣中几乎不表达。实时荧光定量PCR分析结果表明,CaspAG基因在雌雄蕊原基分化期至雌雄蕊成熟期均有表达,在雌雄蕊发育成熟期表达量达到最高;且在雄蕊的表达高峰的时间明显早于雌蕊,这与雌、雄蕊形态成熟的时间基本吻合。
The homology gene cloning technique was used to obtain the AG homologous gene determined by floral organ traits of the golden tree, which was named CaspAG. The open reading frame (ORF) was 738 bp, encoding 245 amino acids. Molecular phylogenetic analysis and protein sequence alignment showed that CaspAG, an AG homologue of Arabidopsis thaliana, was classified as the evolutionary branch of eu AG with 57 amino acids in MADS region, 32 amino acids in I region and 83 in K region The amino acid, C region 55 amino acids, which C-terminal transcriptional activation region contains two conserved motifs: AGI and AGII motif. Semi-quantitative RT-PCR analysis showed that during floral development, the CaspAG gene was only expressed in stamens and pistils but scarcely expressed in stems, leaves, sepals and petals. The results of real-time PCR showed that CaspAG gene was expressed in the primordial period of pistil and stamen to the stamen stage, and reached its peak in the developmental period of the pistil and stamen. The expression peak of stamens was earlier than that of the pistil, This is consistent with the female and stamen morphological maturation time.