月季RcNAC72基因的克隆及表达分析

Gene Cloning and Expression Analysis of Rcnac72 in Rosa chinensis

为揭示月季NAC转录因子的功能,以月季“月月粉”生根的组培苗为材料克隆得到一个NAC基因,命名为RcNAC72。生物信息学分析表明,RcNAC72基因全长为1672 bp,开放阅读框(ORF)为1059 bp,可编码353个氨基酸。基因序列比对发现,RcNAC72在N端具有保守的结构域,与草莓FvNAC72-like、桃PpNAC72、梅花PmNAC72-like、苹果MdNAC91蛋白序列分别有88.34%、80.74%、80.70%、79.96%的相似性。系统进化树分析结果表明,RcNAC72与梅花PmNAC72-like、桃PpNAC72、甜樱桃PaNAC72、樱花PsNAC72,亲缘关系较近。实时荧光定量PCR分析表明,高盐、水杨酸、茉莉酸甲酯均可诱导RcNAC72基因上调表达,高盐,水杨酸,高盐与水杨酸共处理时,根系中的相对表达量明显高于叶片。综上推测RcNAC72基因主要在月季根中作为调节因子参与盐胁迫响应。

In order to reveal the function of the NAC transcription factor in Rosa chinensis,an NAC gene named RcNAC72 was cloned from root fissue culture of Rosa chinensis“Old Blush”.Bioinformatics analysis showed that the full length of RcNAC72 gene was 1672 bp,and the open reading frame(ORF)was 1059 bp,encoding 353 amino acids.Sequence alignment revealed that RcNAC72 had a conserved domain at the N-terminal,and its protein sequences were 88.34%,80.74%,80.70%and 79.96%similar to those of FvNAC72-like,PpNAC72,PmNAC72-like and MdNAC91,respectively.The results of phylogenetic tree showed that RcNAC72 was phylogenetically closed to PmNAC72-like,PpNAC72,PaNAC72 and PsNAC72.Real-time fluorescence quantitative analysis showed that high salt,salicylic acid,methyl jasmonate could induce the increasing expression of RcNAC72.The expression level of RcNAC72 gene in roots was significantly higher than that in leaves.In conclusion,RcNAC72 gene mainly acts as a regulatory factor in roots in response to salt stress.