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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 leaves of lettuce (Lactuca sativa L.) were used as materials, and the sequence of LsHsp70 gene was cloned by RT-PCR and RACE. The heat-resistant variety ’Z36’ and heat-sensitive variety ’S106’ were detected by real- Expression of LsHsp70 gene under different high temperature stress. The results showed that the open reading frame of LsHsp70 gene was 2 094 bp, encoding a protein of 697 amino acids. The molecular weight of LsHsp70 gene was estimated to be 74.1 kD. There are three tagging sequences IDLGTTNS, VFDLGGGTFDVSVL and VILVGGSTRIPAV of HSP70 family with homology of 92% . The response of LsHsp70 to high temperature stress of 37 ℃ was more significant and rapid than that of high temperature stress of 42 ℃ under high temperature stress. Under the same stress temperature, the resistant variety ’Z36’ was more sensitive than the heat-sensitive variety ’S106’ Of the stress response slow, longer duration, indicating that the gene expression and leaf lettuce heat resistance have a certain relevance.