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利用同源克隆技术,在前期课题组对橡胶树转录组进行测序的基础上,根据模式植物中PR1蛋白的氨基酸序列进行同源搜索并设计引物,通过RT-PCR方法扩增巴西橡胶树PR1同源基因的cDNA片段,命名为HbPR1。该片段包含一个450 bp的开放阅读框,编码150个氨基酸的多肽。生物信息学分析结果表明,该氨基酸序列与杨树(Populus tomentosa)、葡萄(Vitis vinifera)和拟南芥(Arabidopsis thaliana)的PR1蛋白高度同源,同源性分别为76%,74%和72%,并含有SCP_PR1_like结构域。半定量分析结果表明,该基因表达受水杨酸的诱导。以上结果暗示该基因为橡胶树的PR1同源基因,可作为橡胶树系统获得性抗性产生的标签基因。
Based on the homology cloning technique, we sequenced the transcriptome of rubber tree from the previous research group and searched for the homology of the PR1 protein in model plants. We designed the primers and amplified the PR1 homologous gene The cDNA fragment, named HbPR1. This fragment contains a 450 bp open reading frame encoding a 150 amino acid polypeptide. Bioinformatics analysis showed that the amino acid sequence was highly homologous to the PR1 protein of Populus tomentosa, Vitis vinifera and Arabidopsis thaliana, with homologies of 76%, 74% and 72%, respectively %, And contains the SCP_PR1_like domain. Semi-quantitative analysis showed that the gene expression was induced by salicylic acid. The above results suggest that this gene is a PR1 homologue of rubber tree, which can be used as a marker gene for acquired resistance to rubbery tree system.