园艺学报 ›› 2013, Vol. 40 ›› Issue (6): 1081-.

• 蔬菜 • 上一篇    下一篇


李 婷,韩莹琰,郝敬虹,范双喜*,任 月   

  1. (北京农学院植物科学技术学院,北京 102206)
  • 收稿日期:2013-02-26 修回日期:2013-05-15 出版日期:2013-06-25 发布日期:2013-06-25

Cloning and Expression Analysis of LsHsp70 from Lactuca sativa  

LI Ting,HAN Ying-yan,HAO Jing-hong,FAN Shuang-xi*,and REN Yue   

  1. (Plant Science and Technology College,Beijing University of Agriculture,Beijing 102206,China)
  • Received:2013-02-26 Revised:2013-05-15 Online:2013-06-25 Published:2013-06-25

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。

关键词: 叶用莴苣, LsHsp70, 克隆, 基因表达, 高温胁迫, 耐热性

Abstract: The full-length cDNA of Hsp70 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 LsHsp70 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.

Key words: Lactuca sativa L., LsHsp70, cloning, gene express, high temperature stress, heat resistance