Abstract

In the current study, the expression levels of mac_4 (HSP80-Like) gene, which is thought to be associated with cold stress in M. piperita, were determined by RT-PCR. First, cold stress-related genes were determined in the model organism Arabidopsis thaliana, since there is no sequence data on cold stress genes in M. piperita, and then BLAST between the two species in the NCBI database was performed to determine the sequences that matched the target genes the most. Primers specific to these sequences were also designed using the NCBI database. In order to determine the correlation of related genes with cold stress in M. piperita plant at the transcriptional level, in vitro grown plants were exposed to cold stress for 24, 72 and 168 hours. To be used in RT-PCR, cDNA was synthesized from total RNAs with reverse transcriptase enzyme. RT-PCR was performed to quantitatively detect the mRNA expression levels of target genes. RT-PCR results were analyzed by the ΔΔCT method. With 18S rRNA normalization, mRNA expression graphs of the gene were generated compared to the control group. TTEST was performed for the significance of the results, and those with p≤0.05 were considered significant. According to the TTEST result, it was observed that there was a significant increase in the mac_4 (Hsp-80 like) gene on cold treated plants.

Keywords: BLAST; Cold stress; Hsp 70; Hsp 90; mac_4 gene