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园艺学报 ›› 2020, Vol. 47 ›› Issue (1): 143-152.doi: 10.16420/j.issn.0513-353x.2019-0306

• 研究报告 • 上一篇    下一篇

芹菜AgHAT4的克隆与表达模式分析

尹 莲1,刘洁霞1,陈龙正2,沈 迪1,段奥其1,冯 凯1,徐志胜1,熊爱生1,*   

  1. 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,农业农村部华东地区园艺作物生物学与种质创新重点实验室,南京 210095;2江苏省农业科学院蔬菜研究所,南京 210014
  • 出版日期:2020-01-25 发布日期:2020-01-25
  • 基金资助:
    江苏省农业自主创新资金项目[CX(18)2007];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 Profiles Analysis of AgHAT4 Gene in Apium graveolens

YIN Lian1,LIU Jiexia1,CHEN Longzheng2,SHEN Di1,DUAN Aoqi1,FENG Kai1,XU Zhisheng1,and XIONG Aisheng1,*   

  1. 1State Key Laboratory of Crop Genetics and Germplasm Enhancement,Ministry of Agriculture and Rural Affairs,Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China,College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;2Institute of Vegetable,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China
  • Online:2020-01-25 Published:2020-01-25

摘要: 以两个芹菜品种‘六合黄心芹’和‘文图拉’为试验材料,分别克隆获得HAT4转录因子基因AgHAT4。应用生物信息学方法分别从氨基酸组成、蛋白结构与理化性质、以及系统进化树等方面对该基因进行分析。结果表明,AgHAT4含有1个长为900 bp的开放阅读框(ORF),编码299个氨基酸。‘六合黄心芹’和‘文图拉’中的AgHAT4基因有1个核苷酸位点的差异,编码的氨基酸有1个位点的差异,其蛋白质相对分子质量分别为33.71和33.68 kD,等电点均为9.12。进化树分析表明芹菜AgHAT4与胡萝卜的HAT4属于同一分支。蛋白结构预测显示,该蛋白主要由3个α螺旋和一些无规则卷曲构成,且未定位到信号肽。荧光定量PCR结果表明,AgHAT4在芹菜叶柄中的表达量最高,根中次之,叶片中表达量最低。多种非生物胁迫导致芹菜AgHAT4的表达量发生变化。与未胁迫对照相比,高温胁迫处理后‘六合黄心芹’中AgHAT4的表达量在24 h明显上调,在盐胁迫条件下‘文图拉’和‘六合黄心芹’中AgHAT4的表达有着相似的变化趋势,均先上升后下降且在处理2 h后表达量达到峰值。‘文图拉’中AgHAT4的表达量在高温处理4 h后表达量最高,而在低温条件下在处理2 h后表达量最高。

关键词: 芹菜, AgHAT4, 基因表达, 组织, 非生物胁迫

Abstract: The transcription factor gene,AgHAT4,was cloned from two celery varieties of‘Liuhe Huangxinqin’and‘Ventura’,respectively. The amino acid composition,protein structure,physicochemical characterizations,and phylogenetic tree were analyzed using bioinformatics methods. Results showed that AgHAT4 contains an open reading frame(ORF)with the length of 900 bp,encoding 299 amino acids. One nucleotide site of AgHAT4 and an amino acid residue were different between the two celery varieties. The relative molecular weights of AgHAT4 protein of‘Liuhe Huangxinqin’and‘Ventura’were 33.71 kD and 33.68 kD respectively,with theoretical pI both about 9.12. Phylogenetic analysis demonstrated that the HAT4 proteins in celery and carrot belonged to the same branch. Protein structure prediction indicated that AgHAT4 protein was mainly composed of three α-helixes and some random coils,and no signal peptide was there. It was detected that AgHAT4 gene was most highly expressed in the petiole,followed by the root and lowest in the leaf blade by real-time quantitative PCR. The results also showed that multiple abiotic stresses induced the expression levels of AgHAT4 in celery. Compared with the control group,the expression levels of AgHAT4 in‘Liuhe Huangxinqin’up-regulated at 24 h under high temperature treatment. The expression response of AgHAT4 to salt stress in‘Ventura’was similar to that in‘Liuhe Huangxinqin’. The transcription levels of AgHAT4 increased first and then decreased in general,which reached the peak after 2 h of treatment. In ‘Ventura’,the expression of AgHAT4 exhibited the highest at 4 h under high temperature treatment,and it exhibited the highest expression at 2 h of low temperature treatment.

Key words: Apium graveolens, AgHAT4, gene expression, tissue;abiotic stress

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