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Role of a serine protease gene (AccSp1) from Apis cerana cerana in abiotic stress responses and innate immunity

  • Lijun Gao
  • Lijun Wang
  • Xinxin Yang
  • Ying Wang
  • Zhenguo Liu
  • Baohua Xu
  • Xingqi Guo
Original Paper

Abstract

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.

Keywords

Apis cerana cerana Clip-domain serine protease Abiotic stresses RNA interference PO activity Antioxidant enzyme 

Notes

Funding

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).

Supplementary material

12192_2018_934_MOESM1_ESM.docx (108 kb)
ESM 1 Survival of adult bees. Groups of 10 adult bees were pretreated with dsRNA 6 h before challengeing Bacillus bombysepticus. One groups were injected 6 μg/individual dsSp1, another groups were injected 8 μg/individual dsSp1. The experiment was repeated three times and gave the same results. (DOCX 107 kb)

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Copyright information

© Cell Stress Society International 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Crop Biology, College of Life SciencesShandong Agricultural UniversityTaianPeople’s Republic of China
  2. 2.College of Animal Science and TechnologyShandong Agricultural UniversityTaianPeople’s Republic of China
  3. 3.College of Life SciencesTaishan Medical UniversityTaianPeople’s Republic of China

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