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园艺学报 ›› 2016, Vol. 43 ›› Issue (11): 2193-2202.doi: 10.16420/j.issn.0513-353x.2016-0462

• 蔬菜 • 上一篇    下一篇

芹菜AgERF4 转录因子基因的克隆与表达分析

吴 蓓,李梦瑶,王广龙,黄 蔚,王 枫,熊爱生*   

  1. 南京农业大学园艺学院,作物遗传与种质创新国家重点实验室,农业部华东地区园艺作物生物学与种质创制重点
    实验室,南京210095
  • 出版日期:2016-11-25 发布日期:2016-11-25
  • 基金资助:

    国家自然科学基金项目(31272175);江苏省杰出青年基金项目(BK20130027);江苏高校优势学科建设项目(PAPD)

Cloning and Expression Profile Analysis of AgERF4 Transcription#br# Factor Gene from Apium graveolens

WU Bei,LI Meng-yao,WANG Guang-long,HUANG Wei,WANG Feng,and XIONG Ai-sheng*   

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement,Ministry of Agriculture Key Laboratory of Biology
    and Germplasm Enhancement of Horticultural Crops in East China,College of Horticulture,Nanjing Agricultural
    University,Nanjing 210095,China
  • Online:2016-11-25 Published:2016-11-25

摘要:

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。

关键词: 芹菜, 乙烯反应元件结合蛋白, 酵母单杂交, 非生物胁迫, 基因表达

Abstract:

In this study,the AgERF4 gene was cloned from celery variety‘Liuhehuangxinqin’.
Sequence analysis indicated that the length of AgERF4 gene was 597 bp,which encoded 198 amino acids.
It is predicted that the molecular mass of its protein was 21.43 kD,and pI was 8.51. Phylogenetic analysis
indicated that the ERF4 showed high evolutionary conservation among celery and other species. The
AgERF4 belongs to hydrophilic protein. Yeast one-hybrid and β-galactosidase activity assays confirmed
the GCC box-binding activities of AgERF4 factor. RT-qPCR analysis demonstrated that the AgERF4 gene
was mainly expressed in leaf blade. The expression profiles of the AgERF4 gene in root and petiole have
no significant difference. When exposed to high or low temperature treatments,the expression level of
AgERF4 gene showed an increased trend at first,followed by a reduction. Under salt treatments,the
transcription of AgERF4 was higher than that of the control. The results suggested that AgERF4 factor
may play important roles in the defense response to abiotic stress in celery.

Key words: Apium graveolens, ERF protein, yeast one-hybrid, abiotic stress, gene expression

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