棉花GhSRO04/GhSRO08基因的克隆及表达分析

Cloning and Expression Analysis of Two SRO Genes in Gossypium hirsutum L.

SRO转录因子在植物响应盐、旱和病菌等生物和非生物胁迫中具有重要的调节作用。本研究从抗枯萎病耐黄萎病的棉花品种鲁研棉32号中克隆获得2个基因:命名为GhSRO04和GhSRO08(Gen Bank登录号为KR534896和KR534895)。结果显示:GhSRO04包含一个最大开放阅读框1830 bp(ORF),推测编码609个氨基酸;GhSRO08包含一个最大开放阅读框1 842 bp(ORF),推测编码编码613个氨基酸。比对分析表明GhSRO04和GhSRO08基因均含有RCD1-SRO-TAF4结构域(RST)。系统进化树分析表明,GhSRO04和GhSRO08编码产物与拟南芥RCD1和SRO1亲缘关系最近,属于同一进化分支。实时荧光定量PCR(q RT-PCR)分析基因表达显示,GhSRO04和GhSRO08基因均受黄萎病菌侵染、干旱和高盐诱导表达,表明GhSRO04和GhSRO08在棉花应对盐、旱和病菌胁迫中可能起着重要的调节作用,对棉花种质的抗逆性遗传改良具有潜在利用价值。本研究首次从棉花中分离鉴定出SRO类转录因子,并进行了初步功能分析,为棉花遗传改良储备了基因资源。

The SIMILAR TO RADICAL-INDUCED CELL DEATH 1(SRO) proteins are important regulatory factors involving in plant responses to biotic and abiotic stresses.Based on genomic sequences of diploid cotton Gossypium raimondii,we cloned two new cotton SRO genes,SRO04 and SRO08,from Lumianyan32,a transgenic Bt upland cotton variety characterized by Fusarium wilt resistance and Verticillium wilt tolerance.Sequence analysis revealed that the open reading frames of GhSRO04 and GhSRO08 are 1 830 bp and 1 842 bp by length,encoding a protein of 609 and 613 amino acids respectively.Compared to other SRO protein sequences,GhSRO04 and GhSRO08 show highest similarity to RCD1 and SRO1 in Arabidopsis thaliana,with highly conserved RCD1-SRO-TAF4 domain.Further analysis by real time quantitative PCR demonstrated that both GhSRO04 and GhSRO08 were induced expressed under stress conditions such as Verticillium strain infection,drought(PEG),and high salt(NaCl),suggesting that these genes may play an important regulatory role in cotton responses to both biotic and abiotic stresses,with the potential value in cotton stress resistance improvement.This is the first report of SRO transcription factors identified in cotton,which provide important gene resources for the genetic improvement of cotton germplasm.