Abstract
Serine proteinases play important roles in innate immunity and insect development. We isolated a serine proteinase gene, designated AccSp10, from the Chinese honeybees (Apis cerana cerana). RT-qPCR and a Western blot analysis at different pupal development stages indicated that AccSp10 might be involved in melanin formation in pupae and promote pupal development. In adult workers, the expression of AccSp10 was upregulated by treatments mimicking harmful environments such as the presence of Bacillus bombysepticus, different temperatures (4, 24 and 42 °C), HgCl2, H2O2 and paraquat; the exception was treatment with VC (vitamin C), which did not upregulate AccSp10 expression. Western blot confirmed the results. A disc diffusion assay indicated that recombinant AccSp10 accelerated E. coli cell death during stimulation with harmful substances (HgCl2, paraquat and cumene hydroperoxide). These findings suggest that AccSp10 may be involved in the pupal development of Chinese honeybees and protection against microorganisms and abiotic harms.
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Acknowledgments
This work was financially supported by the earmarked fund for China Agriculture Research System (no. CARS-45), the National Natural Science Foundation of China (no. 31572470), Shandong Province Modern Agricultural Technology System Innovation Team Special Fund (SDAIT-24-04) and Shandong Province Agricultural Fine Varieties Breeding Projects (2014–2016).
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Fig. S1
A MALDI-TOF/TOF MS of recombinant AccSp10 protein. a A SDS-PAGE of the recombinant AccSp10 protein. 1 and 2 lanes, suspension of sonicated recombinant AccSp10 protein (10 μL) and purified recombinant AccSp10 protein (5μg). The target strip is indicated with arrows. b Mascot Score Histogram and Concise Protein Summary Report. c Matched peptides shown in bold red. (DOCX 358 kb)
Fig. S2
The antibacterial activity locus of the AccSp10 protein. The antibacterial activity loci of the AccSp10 protein were predicted using the APD database. Two antibacterial activity loci are shown in the tertiary structure of activated AccSp10 built using homology modelling in SWISS-MODEL. a The first antibacterial activity locus (from A145 to A182) is shown in blue. b The second antibacterial activity locus (from L326 to T371) is shown in red. (DOCX 322 kb)
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Gao, L., Wang, H., Liu, Z. et al. The initial analysis of a serine proteinase gene (AccSp10) from Apis cerana cerana: possible involvement in pupal development, innate immunity and abiotic stress responses. Cell Stress and Chaperones 22, 867–877 (2017). https://doi.org/10.1007/s12192-017-0818-5
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DOI: https://doi.org/10.1007/s12192-017-0818-5