红掌AP2类转录因子AaSNB编码基因的克隆及表达分析

Cloning and Expression Analysis of AaSNB Encoded Gene of AP2-Type Transcription Factor from Anthurium

红掌是重要的高档盆栽花卉。挖掘红掌观赏器官的关键控制基因,对于利用生物技术改良红掌品质意义重大。本研究采用RACE技术,克隆了一个AP2类转录因子的核苷酸序列,该序列全长2 201 bp,编码区长度为1 461 bp,预测编码486个氨基酸残基。通过NCBI数据库比对和生物信息学分析,发现该基因由两个较为保守的AP2结构域组成,该结构域前端连接有一段核定位信号,在预测氨基酸的氮端具有转录激活功能信号,认定该基因与水稻苞片控制基因SNB为同源类似物,将其命名为AaSNB。采用荧光定量RT-PCR方法初步分析了该基因在红掌不同幼嫩器官的表达情况,发现该基因在肉穗花序中表达量最高,苞片中表达量次之,在根系中也有极微量表达。综合分析认为该转录因子对于红掌苞片等的发育起到重要作用,讨论了利用该基因创制多苞片红掌新种质的可能性。本研究对发掘红掌苞片发育的关键控制基因,并利用其改良红掌品质提供了更多选择,对于丰富AP2转录因子家族也具有积极意义。

Anthurium is an importa nt high-grade potted flower.It is of great significance for improving the quality of Anthurium by biotechnology to dig out the key control genes of the ornamental organs of Anthurium.In this study,the nucleotide sequence of a AP2 type transcription factor was cloned using RACE technology.The full length of the sequence was 2 201 bp and the coding region was 1 641 bp long,which was predicted to encode 486 amino acid residues.Through NCBI database and bioinformatics analysis,it was found that the gene was composed of two relatively conservative AP2 domains,and the front end of this domain was connected with a nuclear location signal.Besides,a transcription activation signal was found at the nitrogen end of this predicted amino acid sequence.Thus,the gene was identified as a homologous analogue to the bract control gene SNB in rice and named AaSNB.The gene expression patterns in different young organs were analyzed by fluorescence quantitative RT-PCR method.The result indicated that the expression level in inflorescence was the highest,followed by the bract,and the content in root was very little.Overall,data analysis showed that this transcription factor played important roles in the development of bract.The possibility of using this gene to create supernumerarybract Anthurium was discussed.This study could provide more options for exploring key control genes of Anthurium to improve its ornamental quality,and could positively enrich the family members of AP2 transcription factor as well.