黄瓜ABC转运蛋白基因(abca19)的克隆及其对农药霜霉威胁迫的响应  

Cloning of ABC Transporter Protein(abca19) in Cucumis Sativus L. and Response under Propamocarb Stress

作　　者：吴鹏[1] 郭茜茜[1] 武涛[2] 秦智伟[2] 

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [2]东北农业大学园艺学院,黑龙江哈尔滨150030

出　　处：《作物杂志》2015年第3期45-51,共7页Crops

基　　金：国家自然科学基金(31272158);齐齐哈尔大学大学生创新创业训练计划(201410221084)

摘　　要：ABC转运蛋白基因与植物的多抗耐药性密切相关,是植物抗农药机理研究的热点。克隆黄瓜ABC转运蛋白基因abca19,分析其编码蛋白的氨基酸序列、理化性质和结构,了解其在霜霉威胁迫条件下的表达规律,为abca19的研究奠定基础,为黄瓜抗农药霜霉威机理研究积累材料。以D0351为材料,根据黄瓜基因组数据库中Csa7M368750.1基因编码区全序列,应用Primer Premier 5.0软件设计引物,从黄瓜果实cDNA中克隆abca19基因的开放阅读框。定量PCR分析霜霉威胁迫处理条件下,黄瓜果实不同处理时间和黄瓜不同部位abca19的表达变化。abca19序列长度为921bp,注册到Gene Bank,登录号为KC123181。abca19编码306个氨基酸,与大豆（Glycine max）等植物abca19的同源性为80%。该蛋白是一个疏水蛋白,无跨膜结构,包含6个α-螺旋结构,无信号肽。具有蛋白激酶C磷酸化位点、N-豆蔻酰化位点、N-糖基化位点和酪蛋白激酶Ⅱ磷酸化位点等多个活性位点。霜霉威处理条件下,0.5-6h黄瓜果实处理组abca19相对表达量极显著高于对照,9-48h对照组abca19相对表达量高于处理组,但二者间差异不显著。不同处理时间点（3、9和24h）,叶片中abca19相对表达量最高,果实中其次,茎中最低。成功克隆到黄瓜abca19,该基因具有已知物种abca19的特征。霜霉威处理条件下,黄瓜果实abca19参与农药霜霉威胁迫响应,反应迅速且具有时限性。在黄瓜叶片和果实中,abca19积极参与对农药霜霉威胁迫的响应,茎中响应较弱。ABC transporter protein is closely related to the plant multiple anti-drug resistances, and is a research hotspot on mechanism of plant resistance to pesticides. We have cloned the cucumber ABC transporter protein gene （abca19） , analyzed its coding protein, physical and chemical properties and structure of the sequence of amino acids,known the abca19 expression pattern under propamocarb stress,laid the foundation for the abca19 research,and accumulated materials for studying cucumber resistance mechanism under propamocarb stress. In this study, D0351 was used as material and we cloned the open reading frame of cucumber ABC transporters genes （ abca19 ） based on the Csa7M368750.1 gene coding region in the cucumber genome database using Primer Premier 5.0. Quantitative PCR was done to analyzed abca19 expression changes under propamocarb stress in different processing time and positions of cucumber fruit. The cucumber abca19 is 921bp. The abca19 is registered to Gene- Bank,and accession number is KC123181. The abca19 encoded 306 amino acids, and analysis of the nucleotide sequence by BLAST indicated that homology of abca19 was 80% with Glycine max. The protein encoded by this gene was a hydrophobic protein with protein kinase C phosphorylation site, N-myristoylation site,ATP/GTP-bind- ing site motif A （P-loop） ,N-glycosylation site, Casein kinase Ⅱ phosphorylation site, and other active sites. This protein contained 6 or-helix structure, and had no transmembrane structure and signal peptides. Under propamocarb stress, the abca19 relative expression in cucumber fruit treatment group within 0.5 - 6h was significantly higher than control. The abca19 relative expression of control group within 9 -48h was higher than the treatment group, but the diffe-rence was not significant. The abca19 relative expression of leaves was the highest, followed by fruits, and stems was the lowest in the different treatment time points （3,9 and 24h.）.. We successfully cloned the cucumber abca19 ,the gene had the characte

关 键 词：黄瓜 abca19 克隆 序列分析 霜霉威 胁迫响应 

分 类 号：S642.2[农业科学—蔬菜学]