Molecular identification of three novel glutaredoxin genes that play important roles in antioxidant defense in Helicoverpa armigera |
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| Affiliation: | 1. Agriculture and Agri-Food Canada, Saskatoon, SK, Canada;2. Department of Biology, University of Saskatchewan, Saskatoon, SK, Canada;3. Department of Plant Protection, College of Agriculture, University of Ankara, Ankara, Turkey;4. Department of Food and Bioproduct Sciences, University of Saskatchewan, Saskatoon, SK, Canada;1. Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA;2. Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA;1. Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;2. Sericultural Research Institute, Jiangsu University of Science and Technology, Zhengjiang 212018, Jiangsu, China;3. Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhengjiang 212018, Jiangsu, China;1. Department of Bio and Fermentation Convergence Technology, BK21 PLUS Project, Kookmin University, Seoul 02707, South Korea;2. Department of Molecular Biology, Chonbuk National University, Jeonju, Jeollabukdo 54896, South Korea;1. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China;2. Environmental Monitoring Center Station, DaQing Environmental Protection Agency, Daqing 163316, China;3. The Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Sciences and School of Life Sciences, South China Normal University, Guangzhou 510631, China |
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| Abstract: | Glutaredoxins (Grxs), also known as thioltransferases, play key roles in maintaining intracellular redox balance and protecting cells from oxidative damage in plants and mammals. We tested whether Grxs play important roles in antioxidant defense in insects using the moth, Helicoverpa armigera. We obtained the full-length cDNA sequences of three novel Grx genes, named HaGrx, HaGrx3, and HaGrx5. Sequence analysis indicated that HaGrx shared a high amino acid identity (58%–78%) and a CPYC motif of conserved redox activity with homologues from other selected insect species. In contrast, HaGrx3 and HaGrx5 both shared a CGF(S/G) motif, a conserved catalytic domain, with other orthologous genes. Quantitative real-time PCR results revealed that HaGrx, HaGrx3, and HaGrx5 exhibited temporally- and spatially-dependent patterns of expression. The mRNA expression of HaGrx, HaGrx3, and HaGrx5 was induced by various temperature stresses and H2O2 treatments. We further investigated the knockdown of HaGrx, HaGrx3, and HaGrx5 in H. armigera larvae and found that most of the selected antioxidant genes were up regulated. However, Tpx was down regulated, and further interpretation of the complementary functions of these antioxidant genes is still required. We also determined the effect of HaGrx, HaGrx3, and HaGrx5 knockdown on antioxidant enzymatic activity and metabolite content. The enzymatic activities of SOD, CAT, and POD, and the metabolite contents of hydrogen peroxide, ascorbate, protein carbonyl, and total GSH increased after RNAi mediated knockdown of HaGrx, HaGrx3, and HaGrx5. These results supported our hypothesis that HaGrx, HaGrx3, and HaGrx5 play important roles in antioxidant defense of Helicoverpa armigera and provided a theoretical basis for further in-depth study of physiological function in the insect glutaredoxin family genes. |
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| Keywords: | Glutaredoxin RNA interference Antioxidant defense Enzymatic activity Metabolite content |
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