论文部分内容阅读
采用同源克隆技术,从黄金树(Catalpa speciosa)花芽中克隆得到B类MADS-box基因Casp AP3和Casp PI的c DNA序列。序列分析表明,Casp AP3基因c DNA序列的完整开放阅读框(ORF)为696 bp,编码231个氨基酸残基;Casp PI基因c DNA序列的ORF为639 bp,编码212个氨基酸残基。蛋白质序列相似性比对和分子系统发生分析表明,Casp AP3属于AP3/DEF进化支,其C末端包含保守的eu AP3基序和PI-derived基序,而Casp PI聚类于PI/GLO进化支,其C末端包含保守的PI基序。半定量RT-PCR分析结果表明,Casp AP3和Casp PI基因均仅在花瓣和雄蕊中表达。实时荧光定量PCR分析表明,Casp AP3和Casp PI基因在花瓣和雄蕊原基分化期至成熟期均有表达,这2个基因在雄蕊中表达高峰出现的时间均早于花瓣;且花瓣中的Casp AP3和Casp PI基因表达高峰均出现在快速伸长阶段;这与花瓣和雄蕊的形态发育阶段相吻合。
Using homologous cloning technique, the C DNA sequences of B class MADS-box genes Casp AP3 and Casp PI were cloned from flower buds of Catalpa speciosa. Sequence analysis showed that the complete open reading frame (ORF) of cp sequence of Casp AP3 gene was 696 bp, encoding 231 amino acid residues. The ORF of cp sequence of Casp PI gene was 639 bp, encoding 212 amino acid residues. Protein sequence similarity analysis and molecular phylogenetic analysis showed that Casp AP3 belongs to the AP3 / DEF clade and contains a conserved eu AP3 motif and a PI-derived motif at its C-terminus, while Casp PI is clustered in the PI / GLO clade , The C-terminal contains a conserved PI motif. Semi-quantitative RT-PCR analysis showed that both Casp AP3 and Casp PI genes were expressed only in petals and stamens. Real-time PCR analysis showed that both Casp AP3 and Casp PI genes were expressed in the petal and stamen primordial stages from differentiation to maturation, and the expression peak of these two genes appeared earlier in the stamens than in the petals. Casp The peak of expression of AP3 and Casp PI genes appeared in the rapid elongation stage, which coincided with the morphological developmental stages of petals and stamens.