黄毛草莓丝氨酸/苏氨酸基因FnSTPK1和启动子克隆及表达分析

Cloning and Expression Analysis of Serine/Threonine Gene FnSTPK1 and Promoter in Fragaria nilgerrensis Schltdl.

为探究丝氨酸/苏氨酸基因STPK1在黄毛草莓(Fragaria nilgerrensis Schltdl.)抗炭疽病中的作用,采用RT-PCR技术从黄毛草莓中克隆了FnSTPK1基因的cDNA(GenBank登录号:MN709781)及其启动子序列(GenBank登录号:MN709783),序列分析结果表明:FnSTPK1基因开放阅读框长为1581 bp,编码526个氨基酸,包含1个STKc结构域,该基因起始密码子上游启动子pFnSTPK1序列为752 bp,预测包含TATA-box、CAAT-box、激素响应元件、光响应元件、环境胁迫顺式作用元件等。同源性分析结果显示:FnSTPK1基因编码的氨基酸序列与森林草莓(Fragaria vesca)编码的氨基酸序列相似性为97.92%。RT-qPCR结果表明,黄毛草莓叶片接种胶孢炭疽菌(Colletotrichum gloeosprioides)后,FnSTPK1基因在接种后不同时间点的相对表达量均显著增加,且在接种48 h时达到最高值,为0 h的10.3倍;黄毛草莓叶片在外源喷施水杨酸(SA)12 h时,FnSTPK1基因的相对表达量达到峰值,为0 h的2.9倍。因此,FnSTPK1可能在草莓抗炭疽病和SA诱导等过程中发挥重要的作用。本研究为进一步探究STPK1基因在野生黄毛草莓抗炭疽病中的功能提供了参考。

In order to explored the expression regulation function of serine/threonine gene FnSTPK1,the cDNA(GenBank accession number:MN709781) and promoter sequence(GenBank accession number:MN709783) of FnSTPK1 gene were cloned from Fragaria nilgerrensis Schltdl.by RT-PCR.Sequence analysis showed that the length of the open reading frame of FnSTPK1 was 1 581 bp,encoding 526 amino acids,and containing a conserved STKc domain.The promoter p FnSTPK1 upstream of the start codon was 752 bp in length,which was predicted to contain TATA-box and CAAT-box,hormone responsive elements,light responsive elements,environmental stress cis-acting elements,etc.The result of homology analysis showed that the amino acid sequence encoded by the FnSTPK1 was 97.92% similar to that encoded by Fragaria vesca RT-qPCR analysis showed that the relative expression level of FnSTPK1 was significantly higher than that of the control group after inoculation with Colletotrichum gloeosprioides on Fragaria nilgerrensis leaves,and reached the highest value at 48 h after inoculation,over 10.3-fold than 0 h;When Fragaria nilgerrensis leaves was treated by SA at 12 h,the relative expression level of FnSTPK1 reached its peak level,over 2.9-fold than 0 h.Therefore,FnSTPK1 may play an important regulatory function in the process of resistance to anthracnose and SA induction of strawberry.This study provide a scientific basis for further investigates the function of STPK1 confers resistance to strawberry anthracnose.