胡萝卜ACC合成酶基因DcACS的克隆与表达分析  

Cloning and Expression Profile Analysis of a DcACS GeneEncoding ACC Synthase From Daucus carota L.

作　　者：王广龙 却枫[2] 陈伯清 任旭琴 熊爱生 WANG Guanglong;QUE Feng;CHEN Boqing;REN Xuqin;XIONG Aisheng(School of Life Science and Food Engineering,Huaiyin Institute of Technology,Huaian,Jiangsu 223003;Key Laboratory of Biology and Germplasm Enhancement of Horticulture Crops in East China,Ministry of Agriculture/State Key Laboratory of Crop Genetics and Germplasm Enhancement/College of Horticulture,Nanjing Agricultural University,Nanjing,Jiangsu 210095)

机构地区：[1]淮阴工学院生命科学与食品工程学院,江苏淮安223003 [2]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,江苏南京210095

出　　处：《核农学报》2018年第12期2326-2334,共9页Journal of Nuclear Agricultural Sciences

基　　金：江苏省自然科学基金青年基金项目(BK20170460);淮阴工学院博士科研启动基金(Z301B16531);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(BK20130027)

摘　　要：为分析ACS基因在胡萝卜不同组织和逆境胁迫条件下的表达情况,以胡萝卜品种黑田五寸为试验材料,克隆得到编码1-氨基环丙烷-1-羧酸合成酶(ACS)的基因DcACS,采用DNAMAN、NCBI、ExPASy和MEGA 5.1等生物信息学软件对其序列特征进行分析,并利用实时荧光定量PCR检测其在不同胡萝卜组织和非生物胁迫条件下的表达水平。序列分析结果表明,胡萝卜DcACS基因全长1 485 bp,编码494个氨基酸;推测的氨基酸序列含有ACS特有的7个保守结构域和4个不变氨基酸残基,在进化关系上与葫芦科的黄瓜和香瓜亲缘关系最接近。实时荧光定量PCR结果表明,DcACS基因在叶片中的表达量最高,具有明显的组织特异性,并响应高温、低温、干旱和盐胁迫等非生物胁迫。本研究克隆的DcACS基因为研究胡萝卜生长发育及响应非生物胁迫等过程提供了一定的理论依据。To investigate the expression profiles of DcACS gene in different carrot tissues and abiotic stresses, a gene DcACS that encodes 1-aminocyclopropane-1-carboxylic acid synthase（ACS） was cloned from carrot cultivar ‘Kurodagosun＇. Bioinformatic tools including DNAMAN, NCBI, ExPASy and MEGA 5.1 were used to analyze the sequence information, and quantitative real-time PCR（RT-qPCR） was performed to detect the expression levels of DcACS gene in carrot tissues and under abiotic stresses. Sequence analysis showed that the DcACS gene was 1 485 bp in length and encoded 494 amino acids. The deduced amino acid sequence contains 7 conserved domains and four invariant amino acid residues unique to ACS. The evolutionary relationship of DcACS was more close to cucumber and melon from the Cucurbitaceae family. RT-qPCR analysis revealed that DcACS gene had the highest expression in leaves, showing obvious tissue specificity. Moreover, this gene could respond to abiotic stresses, such as high temperature, low temperature, drought, and salt stress. The DcACS gene cloned in this study can shed light on studies about carrot in response to plant growth and abiotic stresses.

关 键 词：1-氨基环丙烷-1-羧酸合成酶 基因克隆 非生物胁迫 表达分析 胡萝卜 

分 类 号：S631.2[农业科学—蔬菜学]