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以‘长富2号’苹果为试验材料,从其短枝顶芽中克隆得到1个赤霉素信号转导因子MdGAMYB,对其进行生物信息学和表达分析。结果表明,MdGAMYB的开放阅读框(ORF)长度为1 656bp,编码551个氨基酸,蛋白质分子量为59.741 kD。生物信息学分析表明MdGAMYB编码的蛋白存在多个糖基化位点和磷酸化位点;序列分析表明,Md GAYMB和其他物种的GAMYB蛋白有很高的相似性,均含有保守的R2R3 DNA结合域和GAMYB家族所特有的Box1,Box2和Box3保守区域;系统进化分析表明,Md GAYMB与梨、梅花、草莓、枣和葡萄等的GAMYB蛋白具有较高的同源性。实时荧光定量PCR分析表明,Md GAYMB具有组织表达特异性,在叶片、花和芽中的表达量较高。外源GA3处理抑制了花芽孕育和翌年成花,抑制MdGAMYB的表达。在易成花品种‘烟富6号’中的表达量高于难成花品种‘长富2号’。
Taking ’Changfu No.2’ apple as experimental material, one gibberellin signal transduction factor MdGAMYB was cloned from the shoots of the shoots, and its bioinformatics and expression analysis was carried out. The results showed that the open reading frame (ORF) of MdGAMYB was 1 656 bp in length and encoded 551 amino acids with a molecular weight of 59.741 kD. Bioinformatics analysis showed that MdGAMYB encoded multiple glycosylation sites and phosphorylation sites. Sequence analysis showed that Md GAYMB shared high similarity with GAMYB proteins of other species, and all contained conserved R2R3 DNA binding domains And the conserved regions of Box1, Box2 and Box3 peculiar to the GAMYB family. Phylogenetic analysis showed that Md GAYMB shared high homology with GAMYB proteins such as pear, plum, strawberry, date and grape. Real-time PCR analysis showed that Md GAYMB has tissue-specific expression and higher expression in leaves, flowers and buds. Exogenous GA3 treatment inhibited embryogenesis and flowering the following year, inhibiting the expression of MdGAMYB. The expression level of Yifu 6 in Yichenghua was higher than that of Yulifu No.2, which was difficult to flower.