花烟草NaERF1基因的克隆及在非生物胁迫下的表达模式分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0550

摘要/Abstract

摘要： AP2/ERF类转录因子,是植物所特有的最大的一类转录因子家族,在植物的生长发育过程中,扮演着重要的角色。探究花烟草ERF转录因子的生理功能,为花烟草抵御逆境的分子机制研究提供借鉴。采用同源克隆的方法进行基因克隆。通过对花烟草进行非生物胁迫,运用qPCR的方法进行基因表达模式分析。从花烟草（Nicotiana alata）中克隆了一个属于ERF家族的基因NaERF1。该基因的开放阅读框全长为819 bp,编码了272个氨基酸。生物信息学分析结果表明,该基因编码的蛋白分子量为30.7 kD,等电点为6.07;具有AP2/ERF类转录因子家族典型的保守结构域;该基因主要定位于细胞质内,并含有多个磷酸化位点。同源性分析的结果显示,NaERF1基因与茄科植物的ERF同源性较高,并且与普通烟草的ERF亲缘关系最近。NaERF1基因的表达具有组织表达特异性,花中表达量最高,茎中次之,根和叶中表达量较低。同时,在高盐、干旱、低温、ABA、低钾及H₂O₂等非生物胁迫下,NaERF1的表达呈现5种模式。其中,对低钾及ABA胁迫的响应强烈。NaERF1基因属于AP2/ERF类转录因子,可能广泛参与了花烟草包括非生物胁迫响应在内的众多生理过程。

关键词: 花烟草, 转录因子, ERF, 生物信息学, 基因克隆, 非生物胁迫

Abstract: AP2/ERF transcription factors,unique in plants and the largest family of transcription factors in plants,play an important role in plant growth and development. Exploring the physiological function of ERF transcription factors in Nicotiana alata provides useful references for the study of molecular mechanism of N. alata in resistance to abiotic stresses. Homologous cloning method was used for gene cloning,and qPCR was conducted to analyze the expression profile of NaERF1 under abiotic stress. NaERF1 belonging to ERF family was successfully cloned with ORF 819 bp,encoding 272 amino acids. Bioinformatics analysis showed that the protein encoded by NaERF1 had a molecular weight of 30.7 kD and an isoelectric point of 6.07. It had a typical conservative domain of AP2/ERF transcription factor family. NaERF1 was mainly located in the cytoplasm and contained multiple phosphorylation sites. The results of homology analysis showed that NaERF1 gene had high homology with ERF gene of Solanaceae plants,and had the closest relationship with ERF of common tobacco（Nicotiana tabacum）. The expression of NaERF1 was tissue-specific,with the highest expression in flowers,the second in stems,and the lower expression in roots and leaves. Meanwhile,the expression of NaERF1presented 5 patterns under abiotic stresses such as high salinity,drought,low temperature,ABA,low potassium and H₂O₂. Among them,the response to low potassium and ABA stress was strong. The results of this study suggest that NaERF1 gene belongs to AP2/ERF transcription factor family and may be widely involved in abiotic stress response in N. alata.

Key words: Nicotiana alata, transcription factor, ERF, bioinformatics, gene cloning, abiotic stress

鲁琳, 赵希胜, 刘桂雲, 刘维东, 王艳婷, 吴玲莉, 任学良, 鲁黎明. 花烟草NaERF1基因的克隆及在非生物胁迫下的表达模式分析[J]. 生物技术通报, 2020, 36(1): 1-8.

LU Lin, ZHAO Xi-sheng, LIU Gui-yun, LIU Wei-dong, WANG Yan-ting, WU Ling-li, REN Xue-liang, LU Li-ming. Cloning and Expression Profile Analysis of NaERF1 Under Abiotic Stresses in Nicotiana alata[J]. Biotechnology Bulletin, 2020, 36(1): 1-8.

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