摘要
[目的]了解高羊茅SRP基因结构与功能以及在多种逆境胁迫下的表达变化。[方法]以高羊茅转录组学测序获得的SRP序列为基础,从高羊茅叶片c DNA中应用3’RACE和5’RACE方法扩增出SRP基因全长c DNA序列。[结果]SRP基因c DNA全长为1 165bp,具有一个完整的长度为855 bp的开放阅读框,编码蛋白为284个氨基酸,包含一个REF结构域。通过生物信息学的方法预测SRP基因的结构及功能发现,Fa SRP与单子叶植物SRP蛋白具有相对较高的序列同源性。应用荧光定量PCR技术分析了SRP基因在低氮、干旱、高热以及高盐逆境胁迫下的表达变化发现SRP基因能够应答低氮、干旱、高温等胁迫,但应答机制不一样,可能是通过不同途径调节植物抗逆性,Fa SRP对高盐胁迫不敏感。[结论]该研究为培育抗旱、耐热、营养高效的高羊茅品种提供候选基因与技术储备。
[Objective] This study aimed to study the structure and functions of SRP gene and variation in its expression under abiotic stresses. [Method] Using the SRP sequence obtained from transcriptome sequencing as the template, the full-length cDNA sequence of SRP gene in Festuca arundinacea was amplified using the 3'RACE and 5'RACE methods. [Result] The cDNA sequence of SRP gene has a full length of 1 165 bp, and it contains an open reading frame in full length of 855 bp. The encoded protein by SRP gene is composed of 284 amino acids, and contains a REF domain. The bioinformatic researches on structures and functions of SRPs show that the SRP gene in Festuca arundinacea (FaSRP) has relatively high ho- mologies with SRPs in monocots. Under low nitrogen, drought, high temperature and high salt stresses, the variations in expression of FaSRP gene were studied using fluorescence quantitative PCR. The results showed that FaSRP gene makes responses to low nitrogen, drought and high temperature stresses, but the relevant response mechanisms are not the same, indicating different pathways regulating re- sistance of plants. The expression of FaSRP gene is insensitive to high salt stress. [Conclusion] This study will provide certain candidate gene and technical reserve for breeding of drought- and high temperature-tolerant, nutritious and highly efficient Festuca arundinacea cultivars.
基金
Supported by National Natural Science Foundation of China(31360576)~~
关键词
高羊茅
SRP基因
克隆
表达分析
Festuca arundinacea
SRP gene
Cloning
Expression analysis