شناسایی و بیان ژن پروتئین‌های شوک حرارتی در سن گندم

The sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), is one of the most destructive pests of wheat and reducing Crop production by feeding on leaves, stems, and grains. One of the significant life stages of this pest is diapause, a period affect by low temperatures in autumn and winter, as well as food scarcity in wheat and barley fields. The sunn pest also undergoes diapause during summer. Despite its global importance, genomic resources for sunn pest are limited. In this study, the genes HSP70 and HSP60 were sequenced and their expression in the whole body of adult insects during active (spring) and diapause (summer, autumn, winter) stages was investigated for the first time. Heat shock proteins, alongside antioxidant systems, act as frontline responders to environmental and internal stresses. To achieve this, RNA was extracted from the whole body of adult insects of both sexes using Tri-Reagent (Sigma). Single-strand cDNA was synthesized using the AddScript cDNA Synthesis Kit (Addbio) following the manufacturer's instructions, and the synthesized cDNA served as a template in polymerase chain reaction (PCR). Specific primers for amplifying these genes were designed using Oligo7 and primer3 plus software. PCR was performed using a thermal cycler, and the final PCR products were sent for sequencing. The target gene sequences were identified using Sanger sequencing. The evolutionary relationship of these genes within this pest species was examined in relation to other insects in the Hemiptera order using phylogenetic tree analysis. Accordingly, phylogenetic results indicated that Halyomorpha halys has the closest molecular evolutionary relationship to E. integriceps. Furthermore, the expression of these genes was quantified using qPCR. Gene expression analysis revealing significant upregulation of HSP70 and HSP60 genes during diapause compared to the active phase, suggesting that these genes facilitate insect survival under stress and diapause conditions, possibly operating through similar mechanisms throughout periods of stress and diapause.