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园艺学报 ›› 2018, Vol. 45 ›› Issue (6): 1125-1135.doi: 10.16420/j.issn.0513-353x.2018-0071

• 研究论文 • 上一篇    下一篇

芹菜NAC转录因子基因AgNAC1的克隆及其对非生物胁迫的响应

段奥其,冯 凯,刘洁霞,徐志胜,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:2018-06-25 发布日期:2018-06-25
  • 基金资助:
    国家自然科学基金项目(31272175);教育部新世纪优秀人才支持计划项目(NCET-11-0670);江苏省自然科学基金杰出青年基金项目(BK20130027);江苏高校优势学科建设项目(PAPD)

Cloning and Response to Abiotic Stress of NAC Transcription Gene AgNAC1 in Apium graveolens

DUAN Aoqi,FENG Kai,LIU Jiexia,XU Zhisheng,and XIONG Aisheng*   

  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:2018-06-25 Published:2018-06-25

摘要: 通过RT-PCR方法,从芹菜‘六合黄心芹’和‘文图拉’中分别克隆获得编码NAC转录因子的基因AgNAC1。利用生物信息学方法分析其编码氨基酸序列组成、蛋白质理化性质、亲缘关系、空间结构等,采用荧光定量PCR技术检测基因在不同非生物胁迫下的表达水平。结果表明:‘六合黄心芹’和‘文图拉’AgNAC1开放阅读框长度均为957 bp,编码318个氨基酸,其蛋白质相对分子量分别为36.89和36.77 kD,理论等电点分别为5.94和5.78。AgNAC1蛋白与不同植物中NAC家族成员同源性比对和进化树分析表明,其与胡萝卜DcNAC属于同一个分支,进化距离最近。蛋白功能域预测显示,AgNAC1有多个α螺旋和β转角二级结构单元。荧光定量PCR结果表明,AgNAC1在芹菜叶中表达量最高,具有组织特异性,同时对高温、低温、干旱和盐胁迫均有响应。‘六合黄心芹’中AgNAC1的表达水平在高温处理24 h达到最高。‘文图拉’中AgNAC1的表达水平在高温、低温及盐处理后2 h和8 h高于对照,呈现先下降后上升的趋势,在干旱处理4 h时表达水平最高。

关键词: 芹菜, NAC转录因子, 表达分析, 叶, 非生物胁迫

Abstract: An AgNAC1 gene encoding NAC transcription factor was cloned from Apium graveolens‘Liuhe Huangxinqin’and‘Ventura’by RT-PCR method,respectively. The amino acid sequences,protein physicochemical properties,phylogenetic relationships and spatial structures of AgNAC1 were analyzed. The relative expression levels of AgNAC1 under different abiotic stresses were detected by quantitative real-time PCR. The results showed that the lengths of open reading frame of AgNAC1 genes in‘Liuhe Huangxinqin’and‘Ventura’were both 957 bp and encoded 318 amino acids. The relative molecular masses of AgNAC1 were 36.89 and 36.77 kD,with theoretical pI about 5.94 and 5.78,respectively. The homologous alignment and phylogenetic analysis of AgNAC1 with other NAC TFs in different plants revealed that AgNAC1 and DcNAC of Daucus carota belong to the same branch of evolutionary distance. AgNAC1 had the closest genetic relation with DcNAC. The prediction of protein functional domain showed that AgNAC1 protein contains multiple α-helix and β-turn secondary structure units. Quantitative real-time PCR analysis indicated that the AgNAC1 was tissue-specific in celery,and it mainly expressed in leaf. In addition,AgNAC1 responded to heat,cold,drought and salt stresses. The expression levels of AgNAC1 in‘Liuhe Huangxinqin’peaked at 24 h under high temperature treatment. The expression levels of AgNAC1 in‘Ventura’were higher than that of the control at 2 h and 8 h under high temperature treatment,low temperature and salt treatments,showing a trend of first decrease and then increase. The expression level of AgNAC1 was the highest at 4 h under drought treatment.

Key words: Apium graveolens, NAC transcription factor, expression analysis, leaf, abiotic stress

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