Cloning and differential expression of three heat shock protein genes associated with thermal stress from the wolf spider Pardosa pseudoannulata (Araneae: Lycosidae)

Abstract

• Pphsp70 , Pphsp83 , and Pphsp90 protect Pardosa pseudoannulata from high temperatures. • Pphsp70 , Pphsp83 , and Pphsp90 have conserved sequences and characteristic motifs. • Pphsp70 expression is higher than that of Pphsp83 and Pphsp90 , regardless of sex. Pardosa pseudoannulata is the main predatory natural enemy of crop pests in a paddy ecosystem. When P . pseudoannulata is exposed to unfavorable temperature conditions, the response of heat shock proteins could resist the damage, and is therefore, conducive to the organism’s rapid adaptation to the surrounding stress environment. In this study, we explored the roles of hsp70 and hsp90 genes in response to heat stress, using the rapid amplification of cDNA ends technique and cloned full-length cDNAs of Pphsp70, Pphsp83 , and Pphsp90 . The mRNA expression levels of the three genes under different temperature stresses (25, 28, 31, 34, 37, 40, and 43 °C) and with different duration stresses (4, 8, 12, 16, and 20 h) were analyzed by quantitative real-time polymerase chain reaction. The full-length cDNA of Pphsp70 , Pphsp83 , and Pphsp90 was 2331 base pair (bp), 2466 bp, and 2663 bp, respectively. Phylogenetic analysis of amino acid sequences of Pphsp70 , Pphsp83 , and Pphsp90 showed that the sequences had high homology with that of other spiders. The mRNA expression of all three genes was extremely significantly up-regulated at 43 °C. Moreover at 43 °C, the expression of all three genes in both female and male spiders at the duration of 4 h was the highest compared to that of other stress duration groups. Therefore, it can be inferred that the three genes of P. pseudoannulata play a crucial protective role in resistance in a high-temperature environment.