苦荞NAC转录因子FtNAC17的鉴定及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2020-0556

摘要/Abstract

摘要：

植物特有的NAC转录因子参与植物生长发育和逆境响应。研究苦荞中NAC转录因子在非生物胁迫中的应答,为阐明基因功能提供理论依据。以苦荞为试验材料,克隆一个NAC家族基因,对其进行生物信息学分析。通过荧光定量PCR技术检测该基因在干旱、低温、盐、茉莉酸甲酯、脱落酸和赤霉素胁迫时的响应情况。该基因编码272个氨基酸,将其命名为FtNAC17,GenBank登录号为MT641452。基因结构分析表明,FtNAC17由2个外显子和1个内含子组成。氨基酸序列多重比对和进化关系分析表明,FtNAC17蛋白与拟南芥ANAC002亲缘关系最近,属于NAC转录因子家族中同一亚组。FtNAC17对干旱、低温、盐等非生物胁迫均有不同程度的响应。

关键词: 苦荞, FtNAC17, 克隆, 表达, 非生物胁迫

Abstract:

Plant specific NAC transcription factors are involved in plant growth and development and stress response. To study the response of NAC transcription factors in tartary buckwheat to abiotic stress would provide theoretical basis for elucidating gene function. A NAC family’s gene from tartary buckwheat was cloned and then analyzed by bioinformatics. The response of the gene to drought,low temperature,salt,methyl jasmonate,abscisic acid and gibberellin was detected by fluorescence quantitative PCR. The gene encodes 272 amino acids,which is named FtNAC17 and GenBank accession number is MT641452. The analysis of gene structure showed that FtNAC17 gene was composed of two exons and one intron. Multiple alignment of amino acid sequences and phylogenetic analysis revealed that FtNAC17 protein was the closest relative to ANAC002 in Arabidopsis thaliana,belonging to the same subgroup of NAC transcription factor family. FtNAC17 presented varied responses to drought,low temperature and salt abiotic stress.

Key words: tartary buckwheat, FtNAC17, cloning, expression, abiotic stresses

荣玉萍, 唐彬, 李鹏, 章洁琼, 陈庆富, 朱丽伟, 邓娇, 黄娟. 苦荞NAC转录因子FtNAC17的鉴定及表达分析[J]. 生物技术通报, 2021, 37(1): 174-181.

RONG Yu-ping, TANG Bin, LI Peng, ZHANG Jie-qiong, CHEN Qing-fu, ZHU Li-wei, DENG Jiao, HUANG Juan. Identification and Expression of NAC Transcription Factor FtNAC17 in Tartary Buckwheat[J]. Biotechnology Bulletin, 2021, 37(1): 174-181.

图/表 9

图1 苦荞FtNAC17的扩增 M：Maker;1：以DNA为模板扩增图;2：以cDNA为模板扩增图

图2 FtNAC17的结构

图3 FtNAC17的核酸及其编码的氨基酸序列 ATG：起始密码子;TGA：终止密码子

表1 FtNAC17蛋白的理化性质

基因名称	编码区长度	氨基酸数目	分子量/kD	理论等电点	脂肪族氨基酸指数	蛋白质疏水性
NAC17	816	272	30.77	7.05	63.12	-0.584

表2 FtNAC17蛋白亚细胞定位

位置	可能性/%
微体	64
细胞质	45
线粒体	1

图4 FtNAC17蛋白二级结构预测 h：α-螺旋;e：β-折叠;c：无规则卷曲

图5 NAM保守结构域氨基酸多重序列比对横线表示 A-E亚结构域

图6 FtNAC17与6类NAC蛋白的系统进化分析

图7 FtNAC17在不同胁迫下的表达情况不同小写字母表示差异显著（P < 0.05）

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