地黄RgPR1基因的克隆、生物信息学及表达分析

Cloning,Bioinformatics and Expression Analysis of RgPR1 Gene in Rehmannia glutinosa

基于实验室前期获取的地黄转录组数据,设计了特异性引物,克隆了完整的RgPR1基因序列,通过生物信息学等方法对编码蛋白质进行了分析;利用荧光定量PCR方法分析了植物激素水杨酸和茉莉酸诱导下的RgPR1基因组织差异性表达。结果显示,RgPR1基因全长668 bp,共编码163个氨基酸,其编码的蛋白可能定位于细胞外,为不稳定的分泌蛋白。RgPR1蛋白与一串红的PR1蛋白的同源性最高;经水杨酸和茉莉酸处理后,PR1基因在地黄根、茎、叶中均显著提高,其中在叶中的表达量最高,根中次之,茎中的表达量最低。本研究成功克隆出RgPR1基因,发现其在不同组织中的表达具有一定的差异性,且外源水杨酸和茉莉酸对其表达具有调节作用,为进一步研究PR1基因在地黄抗病中的作用奠定了基础。

Based on the transcriptome data of Rehmannia glutinosa obtained in our lab.The specific primers were designed,and the complete RgPR1 gene sequence of Rehmannia glutinosa was cloned.The bioinformatics analysis was conducted on the protein encoded by RgPR1.The expression level of RgPR1 gene induced by SA and JA on the tissue-specific expression was analyzed.The results showed that the full length of RgPR1 gene was 668 bp,encoding 163 amino acids.The protein encoded by RgPR1 may be located extracellular and is an unstable secreted protein.The RgPR1 protein of Rehmannia glutinosa had the highest homology with the PR1 protein of Salvia splendens.The results of fluorescence quantitative PCR showed that after treatment with SA and JA,the expression of PR1 gene was significantly increased in the roots,stems and leaves of Rehmannia glutinosa,among which the expression level was the highest in the leaves,followed by the roots,and the lowest in the stems.In this study,RgPR1 was cloned,and its expression in different tissues was different.Exogenous SA and JA can regulate the expression of RgPR1,which can lay a foundation for further study on the role of PR1 gene in Rehmannia glutinosa disease resistance.