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以‘六合黄心芹’芹菜为材料,克隆出编码乙烯反应元件结合蛋白基因AgERF4。序列分析表明,AgERF4含有1个长度为597 bp的开放阅读框,编码198个氨基酸,预测其蛋白质相对分子质量为21.43 kD,等电点为8.51。进化分析表明,ERF4转录因子在植物中高度保守。AgERF4属于亲水性蛋白。酵母单杂交和β–半乳糖苷酶活性测定结果确定AgERF4转录因子和乙烯应答元件GCC Box具有较高的结合活性。RT-qPCR分析表明,AgERF4在芹菜叶片中的表达水平较高,在根和叶柄中表达水平较低;高温和干旱条件下AgERF4的表达呈先上升后下降的趋势,在盐处理的24 h内AgERF4的表达始终高于对照。AgERF4转录因子在芹菜非生物逆境胁迫调控过程中起着重要作用。
Using ’Celery’ of ’Liuhe Huangxinqin’ as material, we cloned the AgERF4 gene encoding the ethylene response element binding protein. Sequence analysis showed that AgERF4 contains an open reading frame with a length of 597 bp encoding 198 amino acids. The predicted relative molecular mass of AgERF4 was 21.43 kD and the isoelectric point was 8.51. Phylogenetic analysis showed that ERF4 transcription factor is highly conserved in plants. AgERF4 is a hydrophilic protein. Yeast single hybrid and β-galactosidase activity assay results confirmed that AgERF4 transcription factor and ethylene response element GCC Box have higher binding activity. RT-qPCR analysis showed that the expression level of AgERF4 was higher in celery leaves and lower in roots and petioles. Under high temperature and drought conditions, AgERF4 expression firstly increased and then decreased, and within 24 h after salt treatment AgERF4 expression is always higher than the control. AgERF4 transcription factor plays an important role in the regulation of abiotic stress in celery.