叶用莴苣LsHsp70基因的克隆及表达分析  

Cloning and Expression Analysis of LsHsp70 from Lactuca sativa

作　　者：李婷[1] 韩莹琰[1] 郝敬虹[1] 范双喜[1] 任月[1] 

机构地区：[1]北京农学院植物科学技术学院,北京102206

出　　处：《园艺学报》2013年第6期1081-1089,共9页Acta Horticulturae Sinica

基　　金：北京市自然科学基金项目(6112005);北京市叶类蔬菜创新团队建设专项(blvt-02);北京市科技新星计划项目(2010B020)

摘　　要：以叶用莴苣(Lactuca sativa L.)叶片为试材,采用 RT-PCR 结合 RACE 技术,克隆 LsHsp70基因序列,并利用实时荧光定量 PCR 技术检测耐热品种‘Z36’和热敏品种‘S106’在不同高温胁迫下LsHsp70 基因的表达情况。结果表明,LsHsp70 基因的开放阅读框为 2 094 bp,编码 697 个氨基酸,推测蛋白质分子量为 74.1 kD,具有 HSP70 家族 3 个标签序列 IDLGTTNS、VFDLGGGTFDVSVL 和VILVGGSTRIPAV,与拟南芥 Hsp70 同源性高达 92%。高温胁迫能够诱导 LsHsp70 表达;LsHsp70 对 37 ℃高温胁迫的响应比对 42 ℃高温胁迫的响应更为显著和迅速;在相同胁迫温度下,耐热品种‘Z36’比热敏品种‘S106’对高温的应激反应迟缓,持续时间更长,说明该基因的表达与叶用莴苣耐热性有一定的关联。The full-length cDNA of lisp70 in Lactuca sativa L. was cloned using reverse transcription polymerase chain reaction （RT-PCR） and rapid-amplification of cDNA ends （RACE）. The results showed that LsHsp70 open reading frame is 2 094 bp which encoding a polypeptide of 697 amino acids with an estimated molecular mass of 74.1 kD. The three highly conserved HSP70 family signatures is IDLGTTNS, VFDLGGGTFDVSVL and VILVGGSTRIPAV. The similarity of amino acids between Hsp70 with the homologous gene in Arabidopsis thaliana is 92%. The expression patterns were analyzed by Real-time quantitative PCR in heat-resistant type ＇Z36＇ and heat-sensitive type ＇S106＇ under different high temperatures. The results showed that the high temperature could promote LsHsp70 expression, and LsHspTO expression responses under 37 ℃were more sensitive and had a large accumulation than under 42 ℃ ; In the same temperature, ＇Z36＇ responses speed were more slowly and last longer.

关 键 词：叶用莴苣 LsHsp70 克隆 基因表达 高温胁迫 耐热性 

分 类 号：S636.2[农业科学—蔬菜学]