Role of a serine protease gene (AccSp1) from Apis cerana cerana in abiotic stress responses and innate immunity
Clip-domain serine proteases (Clip-SPs) mediate innate immunity and embryonic development in insects. However, the function of Clip-SPs in Apis cerana cerana is little known. Here, a Clip-SP gene, AccSp1, was identified. AccSp1 was mainly detected in third and sixth day instar larvae, dark-eyed pupae, and adults (1and 30 days post-emergence). In addition, AccSp1 was expressed at its highest level in the venom gland and epidermis than tentacle, abdomen, muscle, honey sac, head, leg, chest, hemolymph, rectum, and midgut. AccSp1 was induced by 4, 24, and 44 °C; H2O2; CdCl2; HgCl2; and pesticides (paraquat, pyridaben, and methomyl) and was inhibited by UV light and cyhalothrin treatments. When adults that had been pretreated with dsRNA 6 h prior (knocking AccSp1 down) were challenged with Bacillus bombysepticus for 18 h, the survival rate of bees greatly decreased, the activity of PO (phenoloxidase) was reduced, revealing that AccSp1 may play a critical role in assisting bees to survive the microbial infection and participate in regulating PO activity. The antioxidant enzymatic activities of catalase, peroxidase, and superoxide dismutase; the contents of hydrogen peroxide and malondialdehyde; and the ratio of NADP+/NADPH were all lower in samples containing dsRNA-AccSp1 interference than in control groups, but the content of carbonyl was not significantly different. These findings suggest the knockdown of AccSp1 may influence melanization so that the antioxidant enzyme activities and the harmful metabolites decreased. These results collectively suggest that AccSp1 plays critical roles in abiotic stresses responses and resistance to pathogens.
KeywordsApis cerana cerana Clip-domain serine protease Abiotic stresses RNA interference PO activity Antioxidant enzyme
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).
- An MI, Choi CY (2010) Activity of antioxidant enzymes and physiological responses in ark shell, Scapharca broughtonii, exposed to thermal and osmotic stress: effects on hemolymph and biochemical parameters. Comparative Biochemistry & Physiology Part B Biochemistry & Molecular Biology 155:34–42CrossRefGoogle Scholar
- Christophides GK, Zdobnov E, Barillas-mury C, Birney E, Blandin S, Blass C, Brey PT, Collins FH, Danielli A, Dimopoulos G, Hetru C, Hoa NT, Hoffmann JA, Kanzok SM, Letunic I, Levashina EA, Loukeris TG, Lycett G, Meister S, Michel K, Moita LF, Müller HM, Osta MA, Paskewitz SM, Reichhart JM, Rzhetsky A, Troxler L, Vernick KD, Vlachou D, Volz J, von Mering C, Xu J, Zheng L, Bork P, Kafatos FC (2002) Immunity-related genes and gene families in Anopheles gambiae. Science 298(5591):159–165CrossRefGoogle Scholar
- Gao L, Wang H, Liu Z, Liu S, Zhao G, Xu B, Guo X (2017) 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 Chaperones 12:1–11Google Scholar
- Greenberger JS, Epperly MW (2004) Radioprotective antioxidant gene therapy: potential mechanisms of action. Gene Ther Mol Biol 8:31–44Google Scholar
- Hallman GJ, Denlinger DL (1998) Temperature sensitivity in insects and application in integrated pest management. Westview Press, Oxford, p 29Google Scholar
- Hartman E (1931) A flacherie disease of silkworm caused by Bacillus bombysepticus. Lignan Science Journal 10:279–289Google Scholar
- Jiang H, Vilcinskas A, Kanost MR (2010) Immunity in lepidopteran insects. Adv Exp Med Biol 34:89–100Google Scholar
- Lin P, Cheng TC, Ma SY, Gao JP, Jin S, Jiang L, Xia QY (2016) Bacillus bombysepticus α-toxin binding to G protein-coupled receptor kinase 2 regulates cAMP/PKA signaling pathway to induce host death. Science Foundation in China 12(2):35–35Google Scholar
- Rawlings ND, Barrett AJ (1993) Evolutionary families of peptidases. J Cell Biochem 290:205–218Google Scholar
- Yoder JA, Denlinger DL (1991) Water balance in flesh fly pupae and water vapor absorption associated with diapause. J Exp Biol 157:273–286Google Scholar