月季RhATAF1基因的克隆及表达特性分析

丁爱琴, 李绍翠, 刘庆超, 王奎玲, 李伟, 刘庆华*, 姜新强*
青岛农业大学园林与林学院, 山东青岛266109

通信作者:刘庆华;E-mail: jiangxinqiang8@163.com, lqh6205@163.com

摘 要:

以切花月季‘萨蔓莎’为材料, 从花瓣水分胁迫抑制消减杂交文库中获得NAC转录因子基因片段, 利用RACE技术分离获得一个新的NAC类型的转录因子, 命名为RhATAF1RhATAF1开放阅读框包含890 bp (登录号为MG878302), 编码289个aa, 由3个外显子和2个内含子组成。生物信息学分析结果显示, RhATAF1蛋白分子量为32.9 kDa, 等电点pI为7.58。RhATAF1二级结构主要以α-螺旋和无规则卷曲为主, β-折叠较少。蛋白序列多重比较发现, RhATAF1具有高度保守的N端DNA结构域及高度分化的C端转录调节区域, N端区域包含了A、B、C、D、E 5个不同的亚结构域。系统发育分析表明RhATAF1与拟南芥ATAF1同源性最高, 属于SNAC类转录因子。利用实时定量PCR技术分析了RhATAF1的表达模式, 结果显示, 水分胁迫12 h处理显著提高了RhATAF1的表达水平, 脱落酸和赤霉素处理诱导了RhATAF1的转录本表达上调。

关键词:切花月季; RhATAF1; 实时定量PCR; 表达特性

收稿:2018-04-05   修定:2018-09-07

资助:山东省优秀中青年科学家科研奖励基金(BS2014SW032)和国家自然科学基金(31501798)。

Cloning and expression analysis of RhATAF1 gene in Rosa hybrida

DING Ai-Qin, LI Shao-Cui, LIU Qing-Chao, WANG Kui-Ling, LI Wei, LIU Qing-Hua*, JIANG Xin-Qiang*
College of Landscape Architecture and Forestry, Qingdao Agriculture University, Qingdao, Shandong 266109, China

Corresponding author: LIU Qing-Hua; E-mail: jiangxinqiang8@163.com, lqh6205@163.com

Abstract:

A new NAC transcription factor gene was cloned from the cut roses Rosa hybrida ‘Samantha’. This gene was named RhATAF1, which obtained from water deficit stress related suppression subtractive hybridization cDNA library in rose petals using RACE technology. RhATAF1 GenBank accession number is MG878302. The open reading frame of RhATAF1 was 890 bp, encoded a polypeptide of 289 amino acids and comprised of 3 exons and 2 introns. Bioinformatics analysis indicated that the theoretical molecular weight of RhATAF1 was 32.9 kDa and an isoelectric point was 7.58. RhATAF1 second structure was manly consist of α-helix, random curls and less β-fold. Multiply sequences alignments revealed RhATAF1 contained highly conserved DNA-binding domain in N-terminal and divergent transcription regulon domain in C-terminal. Five sub-domains including sub-domain A, B, C, D and E were found in N-terminal. Phylogenic analysis indicated that RhATAF1 was most similarly with Arabidopsis thaliana ATAF1, and belongs to the NAC proteins SNAC sub-family. Qualitative RT-PCR results showed that RhATAF1 expression was significantly induced by 12 h water deficit stress treatment. In addition, abscisic acid and gibberellic acid upregulated RhATAF1 transcript expression levels.

Key words: cut rose; RhATAF1; qualitative RT-PCR; expression characteristic

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