Characterization of Three Heat Shock Protein Genes in Pieris melete and Their Expression Patterns in Response to Temperature Stress and Pupal Diapause

SIMPLE SUMMARY: Pieris melete, a major pest of crucifers, undergoes obligatory diapause as pupae to survive unfavorable temperature extremes during hot summers and cold winters. Heat shock proteins 70 (Hsp70) participate in this process; however, little is known about the underlying changes in Hsp70 expression both during the summer and winter diapause. The study aimed to investigate expression patterns of Hsp70s (PmHsc70/PmHsp70a, b) in response to diapause and short-term temperature stresses. The results showed that the expression of PmHsc70 and PmHsp70b were upregulated both in summer and winter diapause. Heat shock significantly induced up-regulation of the three genes in both summer and winter diapause. In non-diapause pupae, none of the genes responded to cold or heat stress. Further, it was found that 39 °C for 30 min was the most sensitive heat stress condition for PmHsc70 expressions in summer diapause and all three genes’ expressions in winter diapause. During summer diapause, the expression of the genes was up-regulated in response to high temperature acclimation at 31 °C. Meanwhile, only PmHsp70a and PmHsp70b were up-regulated when acclimated to a low temperature of 4 °C in winter diapause. In conclusion, the current results indicate that PmHsp70s plays a crucial role during both summer and winter diapause, in response to temperature stresses; and our findings may contribute to the increasing knowledge on seasonal diapause adaption. ABSTRACT: Heat shock protein 70 genes participate in obligatory pupal diapause in Pieris melete to survive unfavorable conditions. In this study, three full-length cDNAs of PmHsc70, PmHsp70a and PmHsp70b were identified, and their expression patterns in response to diapause and short-term temperature stresses were investigated. Summer and winter diapause were induced in the pupae and non-diapause individuals were used as a control. The pupae from each diapause group were subjected to either hot or cold conditions and the expression levels of the HSP genes were measured. Our results showed that up-regulation of PmHsc70 and PmHsp70b were detected both in summer and winter diapause, but not for PmHsp70a. Under cold stress, PmHsp70a and PmHsp70b were upregulated in summer and winter diapause, while heat shock significantly induced upregulation of all three genes. In non-diapause pupae, none of the genes responded to cold or heat stress. Furthermore, we found that incubation at 39 °C for 30 min was the most sensitive heat stress condition for PmHsc70 expression in summer diapause. On the other hand, the same temperature was effective for PmHsc70, PmHsp70a, and PmHsp70b expression in winter diapause. During summer diapause, expression of all three genes was upregulated in response to high-temperature acclimation at 31 °C, but only PmHsp70a and PmHsp70b were upregulated when acclimated to a low temperature of 4 °C in winter diapause. These results suggest that the PmHsc70, PmHsp70a, and PmHsp70b respond differently to pupal diapause and temperature stress, and that PmHsc70 is more sensitive to heat shock than to cold stress.