The role of melatonin and Tryptophan-5-hydroxylase-1 in different abiotic stressors in Apis cerana cerana

Abstract

Tryptophan-5-hydroxylase-1 (T5H-1) is the rate-limiting enzyme in the biosynthesis of serotonin, which is involved in the biosynthesis of melatonin (Mel). Mel, a biological hormone, plays crucial roles in stressors tolerance, such as cold, hot, Ultraviolet (UV) and pesticide tolerance. However, the direct correlation between T5H-1 and Mel and the underlying mechanism in organisms remains elusive. Mel-mediated cold tolerance was studied extensively in plants and somewhat in insects, including bees. The present study isolated the Mel synthesis gene T5H-1 from Apis cerana cerana for the first time. qRT-PCR analysis indicated that AccT5H-1 played vital roles during some adverse conditions, including 4 °C, 8 °C, 10 °C, 45 °C, UV, cyhalothrin, abamectin, paraquat and bifenthrin exposure. Knockdown of AccT5H-1 using RNA interference (RNAi) technology upregulated most antioxidant genes. Additionally, an enzyme activity assay revealed higher contents of Malondialdehyde (MDA) and Hydrogen peroxide (H2O2), lower content of Vitamin C (VC), and higher activities of Glutathione S-transferase (GST), Superoxide dismutase (SOD), Catalase (CAT) and Peroxidase (POD) in the AccT5H-1 silenced group than the control group. These results suggest that AccT5H-1 is involved in the response to different oxidative stressors in A. cerana cerana. The survival rate of A. cerana cerana exposed to low temperature treatment revealed that the optimal concentration of Mel in the diet was 10 µg/mL. We also found that the antioxidant enzyme (GST, SOD, POD and CAT) concentrations at 10 µg/mL Mel increased to different degrees, and the content of oxidizing substances (MDA and H2O2) decreased, the content of VC increased, and the content of substances that promote cold resistance (glycerol and glycogen) increased. Mel increased the resistance of A. cerana cerana exposed to low temperatures. The expression of AccT5H-1 decreased after the feeding of exogenous Mel to bees. These results provide a reference for other insect studies on Mel and T5H-1.