Cloning and expression analysis of two small heat shock proteins genes from Aquilaria sinensis / 中草药

ABSTRACT

Objective: To aim at cloning the open reading frame (ORF) of sHSP1 and sHSP2 genes from Aquilaria sinensis and analyzing the bioinformatics and expression of the two genes. Methods: Two unique sequences containing sHSPs domain were discovered in transcriptome dataset of A. sisnensis. The full-length cDNAs of sHSP1 and sHSP2 were cloned by RT-PCR strategy with the specific primers. Subcellular localization, transmembrane domain, three-dimensional structure, and phylogenetic analysis were predicted by different softwares to analyze the bioinformatics of sHSPs protein. The expression different levels of sHSP1 and sHSP2 isoforms in different tissues and in responds to salt and ABA, SA, MJ treatment were measured by real-time quantitative PCR. Results: The sHSP1 and sHSP2 cDNA sequence consisted of 474 bp ORF, encoding 157 amino acids. Tissue expression analysis indicated that sHSP1 and sHSP2 were primarily expressed in roots, followed by stems and leaves. Salt treatment experiments indicated that salt treatment caused a rapid increase in sHSP1and sHSP2 expression within 36 and 24 h, respectively. Exogenous ABA and MJ treatment experiments indicated that sHSP1 and sHSP2 genes were induced by exogenous ABA and MJ, and all reached the highest expression level at 12 h. Simultaneously, the SA treatment experiments indicated that exogenous SA treatment caused a rapid increase in sHSP1 and sHSP2 expression within 12 and 24 h, respectively. Conclusion: The full-length cDNA sequence of sHSP1 and sHSP2 genes from A. sinensis is obtained. sHSP1 and sHSP2 have the different expression level in different tissues. When subjected to high salt, ABA, SA, and MJ treatment, sHSP1 and sHSP2 show the different expression levels in different time. Cloning and analyzing sHSP1 and sHSP2 genes from A. sinensis will play an important role for further study on plant defense response.