Abstract:  【Aim】 The development of honeybee eggs, larvae and pupae is markedly stenothermic, and low-temperature stress leads to arrested development. The purpose of this study is to investigate the molecular mechanism of pupation resumption in Apis mellifera worker prepupae post lowtemperature stress. 【Methods】 Transcriptomes were compared between pupae workers developed at the optimal temperature of 35 ℃ (CK) and pupae from prepupae recovered from low-temperature stress at 20 ℃ for 24 h (T) of A. mellifera workers. The differentially expressed genes (DEGs) in the CK vs T comparison group were identified and annotated in GO and KEGG databases for enrichment. Five antioxidant-related DEGs (SOD2, Tpx4, GstS1, Prdx6 and Cu-Zn) were validated using RT-qPCR. 【Results】 A total of 1 335 DEGs were identified in the CK vs T comparison group, with 853 DEGs up-regulated and 482 DEGs down-regulated. GO enrichment result revealed that the DEGs were significantly enriched in such GO terms as intracellular ribonucleoprotein complexes, NADH dehydrogenase activity, and antioxidant activity. KEGG enrichment result showed that DEGs were significantly enriched into oxidative phosphorylation, ribosome, and proteasome pathways. The RT-qPCR results confirmed the upregulation of 5 antioxidant-related DEGs (SOD2, Tpx4, GstS1, Prdx6, and Cu-Zn), consistent with the transcriptome sequencing results.【Conclusion】 When recovered from low-temperature stress, the A. mellifera prepupae can resume development to complete pupation by increasing energy metabolism, protein synthesis and degradation, as well as the expression of antioxidant-related genes. This research provides insights into the development resumption mechanisms of stenothermic insects after experiencing low-temperature stress.

Key words: Apis mellifera, low-temperature stress, pupation, antioxidant enzyme, metabolism, transcriptome