摘要
ERF是植物中一类重要的转录因子,广泛参与植物对生物和非生物胁迫的响应。以3年生银杏苗为试材,采用RT-PCR与RACE-PCR方法,研究了银杏ERF基因序列,以期深入了解银杏药用成分合成代谢的分子机理。结果表明:GbERF1含有长度为1 314bp的完整开放阅读框,编码437个氨基酸。蛋白序列分析显示,GbERF1蛋白含有2段ERF家族的特殊位点。GbERF1蛋白序列与北美云杉(ADE75663.1)序列的相似性为97%。系统进化树分析,GbERF1与北美云杉、狭叶羽扇豆的序列可划归为同一分支。实时荧光定量PCR结果显示,GbERF1主要在银杏根和叶中表达,逆境胁迫处理后显示,该基因在乙烯、紫外诱导处理下,表达量大幅度提升,随后下降,但仍高于对照组,干旱诱导处理下表达量先升高后降低,矮壮素(CCC)诱导下表达量变化不太明显。烟草叶片下表皮细胞亚细胞定位结果表明GbERF1定位于细胞核。
ERF transcription factors are important in plants,which are widely involved in plant response to biotic and abiotic stress.Three-year-old Ginkgo biloba L.tree was used as materials,the sequences of ERF was explored by using RT-PCR and RACE-PCR technology,in order to understand the molecular mechanism of medicinal component metabolism of Ginkgo biloba L.The results showed that GbERF1 contained a 1 314 bp ORF encoding 437 amino acid residues.NCBI-BLAST showed GbERF1 protein had two specific hits of ERF superfamily.GbERF1 shared 97% similarity with sequences of Picea sitchensis.Phylogenetic tree analysis showed that GbERF1 was grouped into the same clade with sequences of Picea sitchensis and Lupinus angustifolius.The expression patterns of GbERF1 genes in Ginkgo biloba L.tissues and under stress treatments were analyzed by real-time quantitative PCR,the results showed that GbERF1 genes expressed highly in root and leaf.The expression level of GbERF1 under the treatment of ET and UV was significant higher than that of control group,and expression trend of them was upgrade at first and then descend.The expression trend of them under drought was upgrade at first and then descend,and the expression difference was not significant under CCC treatment.Following transient expression of GbERF1 in epidermal cells of leaf of tobacco,GbERF1 was found to be localized in the nucleus.
出处
《北方园艺》
CAS
北大核心
2018年第3期92-100,共9页
Northern Horticulture
基金
湖北省高端人才引领培养计划资助项目(鄂科技通[2012]86号)
国家自然科学基金资助项目(31270717
31400556)
湖北省自然科学重点资助项目(2013BHE029)
关键词
银杏
ERF转录因子
荧光定量分析
亚细胞定位
Ginkgo biloba L.
ERF transcription factors
fluorescence quantitative analysis
subcellularlocalization
