春尺蠖2个小热激蛋白编码基因的克隆及其在不同时空和温度胁迫下的表达情况OA

This study aimed to clone two small heat-shock protein genes(sHSPs)from Apocheima cinerarius and to ana-lyse their expression patterns across different developmental stages,tissues and temperature stresses,so as to clarify the role of sHSPs in the response of A.cinerarius to temperature stress.Based on the pupal transcriptome of A.cinerarius,two sHSP genes were screened and cloned,and subjected to phylogenetic and domain-related bioinformatic analyses.Quantitative real-time PCR(qPCR)was used to examine the spatiotemporal expression characteristics of the two sHSP genes in different developmental sta-ges(egg,larva,pupa and adult)and tissues(head,thorax,abdomen,legs and wings)of A.cinerarius,and to analyse their ex-pression profiles in 4th-instar larvae exposed to temperatures of-5℃,0℃,5℃,25℃and 30℃.The two cloned sHSP genes were designated AcinHSP20.4 and AcinsHSP(GenBank accession numbers PV053122 and PV053123,respectively);their coding sequences(CDS)were 543 bp and 758 bp in length,encoding 180 and 252 amino acids with predicted molecular masses of 20 470 and 27 758 and theoretical isoelectric points of 6.98 and 4.60,respectively.Expression analysis revealed that AcinHSP20.4 transcript levels were low from the egg to the 5th-instar larval stage and peaked in diapause-terminated pupae,whereas AcinsHSP transcript levels remained low from the egg to the 5th-instar larval stage and reached their maximum in female adults.The relative expression level of AcinHSP20.4was the highest in the thorax of female adults,while that of AcinsHSP was the highest in the head of male adults.As temperature increased,the relative expression of AcinHSP20.4 first increased and then decreased,peaking at 25℃and decreasing significantly at 30℃(P<0.05).The relative expression of AcinsHSP showed an overall upward trend,with no significant difference within the temperature range of 0-30℃,but a significant difference was observed between-5℃and 30℃(P<0.05).These results indicate that sHSP genes play important roles in the growth,development,diapause and temperature-stress response of A.cinerarius,and provide a theoretical basis for further investigation into the mechanisms underlying its development,diapause and adaptation to temperature stress.