蜻蜓凤梨AfERF113基因的克隆与表达特性

发布时间：2018-05-05 08:04

本文选题：蜻蜓凤梨 + AP/EREBP家族　；参考：《西北农业学报》2016年12期

【摘要】：观赏凤梨是一类重要的热带花卉,但有关其生长发育调控分子机理的研究相对匮乏,这在一定程度上限制了新品种的开发和利用。以热带观赏花卉蜻蜓凤梨(Aechemia fasciata)为材料,在分析蜻蜓凤梨转录组数据的基础上,结合cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术克隆了APETALA2/Ethylene Response Element Binding Protein(AP2/EREBP)家族的1个转录因子编码序列,并将其命名为AfERF113。生物信息学分析表明,AfERF113 cDNA全长1 093bp,有1个864bp的开放阅读框(open reading frame,ORF),编码287个氨基酸。蛋白质理化性质分析表明,该蛋白分子质量为29.768 5ku,理论等电点为6.06,为一类不稳定的酸性亲水蛋白。亚细胞定位软件预测表明该蛋白定位于细胞核。结构域分析表明,该蛋白有1个保守的AP2结构域,分属ERF亚族的B-4类,与拟南芥中所有ERF蛋白的ERF113亲源关系最近。实时荧光定量PCR(quantitative real time PCR,qRT-PCR)结果表明,AfERF113转录本在成熟株的根中表达量最高,且外源乙烯诱导后,AfERF113的表达量无论是在幼株还是在成株的各组织中均显著上调。研究结果证实,AfERF113响应了外源乙烯调控,为进一步研究AfERF113基因的功能及通过基因工程手段调控凤梨科植物生长发育提供了理论依据。
[Abstract]:Ornamental pineapple is an important tropical flower, but the molecular mechanism of its growth and development regulation is relatively scarce, which limits the development and utilization of new varieties to a certain extent. Using Aechemia fasciata, a tropical ornamental flower, as a material, based on the analysis of transcriptional data, a transcription factor encoding sequence of APETALA2/Ethylene Response Element Binding protein AP2 / EREBP family was cloned by cDNA terminal rapid amplification of cDNA endsracea technique. And named Aferf113. Bioinformatics analysis showed that the cDNA of AfERF113 was 1 093bp. there was an open reading frame of 864bp, which encodes 287-amino acid. The physicochemical properties of the protein show that the molecular weight of the protein is 29.768 kuand the theoretical isoelectric point is 6.06. it is a kind of unstable acidic hydrophilic protein. Subcellular localization software predicted that the protein was located in the nucleus. Domain analysis showed that the protein had a conserved AP2 domain, belonging to B-4 of ERF subfamily, and had the closest relationship with ERF113 of all ERF proteins in Arabidopsis. The results of real-time PCR(quantitative real time PCR qRT-PCRR showed that the expression of AfERF113 transcripts was the highest in the roots of mature strains, and the expression of AfERF113 was upregulated in both young and adult tissues after exogenous ethylene induction. The results showed that AfERF113 responded to exogenous ethylene regulation, which provided a theoretical basis for further study on the function of AfERF113 gene and regulation of the growth and development of pineapple plants by means of genetic engineering.
【作者单位】：中国热带农业科学院热带作物品种资源研究所农业部华南作物基因资源与种质创新重点实验室;
【基金】：国家自然基金面上项目(31372106) 海南省自然科学基金面上项目(20163127)~~
【分类号】：S682.3

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