Characterization of the insecticide detoxification carboxylesterase Boest1 from Bradysia odoriphaga Yang et Zhang (Diptera: Sciaridae).

BACKGROUND: In insects, carboxylesterases (CarEs) are enzymes involved in the detoxification of insecticides. However, the molecular mechanism of CarE-mediated insecticide metabolism in Bradysia odoriphaga, a serious agricultural pest, remains unclear. The aim of this study is to investigate the detoxification process of malathion, bifenthrin, and imidacloprid by B. odoriphaga carboxylesterase (Boest1). RESULTS: An alpha class CarE gene Boest1 was cloned from B. odoriphaga. The results of real-time quantitative polymerase chain reaction showed that Boest1 is up-regulated with age during the larval stage, and the level of transcription of Boest1 is higher in the midgut and Malpighian tubule than in other tissues. The expression level of Boest1 was significantly increased after exposure to malathion and bifenthrin. Recombinant BoEST1 expressed in vitro showed high catalytic activity toward α-naphthyl acetate, which was substantially inhibited by malathion and triphenyl phosphate. The in vitro metabolism assays showed that BoEST1 demonstrates hydrolytic capacity toward malathion and bifenthrin but not imidacloprid. The binding free energy analysis indicates that BoEST1 has a higher affinity for malathion and bifenthrin than imidacloprid. CONCLUSION: These results suggest that BoEST1 plays a role in the breakdown of insecticides and may be involved in the development of resistance in the Chinese chive pest B. odoriphaga; our findings also provide data for better pest management and perspectives for new pesticides development. © 2021 Society of Chemical Industry.

Identification of a novel cytochrome P450 CYP3356A1 linked with insecticide detoxification in Bradysia odoriphaga.

BACKGROUND: Cytochrome P450 monooxygenases play an important role in the metabolic detoxification of insecticides in insect pests. However, little is known about the role of a specific P450 gene and its responses to insecticide exposure in Bradysia odoriphaga, a major pest in Chinese chive production. RESULTS: In this study, a novel P450 gene, CYP3356A1, was cloned from Bradysia odoriphaga. The full-length cDNA sequence of CYP3356A1 is 2153 bp and its open reading frame (ORF) encodes 508 amino acids. Quantitative real time PCR(qRT-PCR) analyses in different tissues showed that CYP3356A1 expression was the highest in the Malpighian tubule. Moreover, among the different developmental stages of the insect, the highest expression of CYP3356A1 was found in fourth-instar larvae. Expression of CYP3356A1 was upregulated by treatment with imidacloprid, thiamethoxam, and ß-cypermethrin at median lethal concentrations (LC50 ). RNA interference (RNAi)-mediated silencing of CYP3356A1 significantly increased mortality by 36.90%, 25.17%, and 36.73 when fourth-instar B. odoriphaga larvae were exposed to imidacloprid, thiamethoxam, and ß-cypermethrin, respectively, at the LC50 dose. CONCLUSION: These results demonstrate that CYP3356A1 is related to the detoxification of imidacloprid, thiamethoxam, and ß-cypermethrin in B. odoriphaga. Moreover, the study also increased our understanding of the molecular mechanisms of insecticide detoxification in this pest insect. © 2018 Society of Chemical Industry.

New Zealand glowworm (Arachnocampa luminosa) bioluminescence is produced by a firefly-like luciferase but an entirely new luciferin.

The New Zealand glowworm, Arachnocampa luminosa, is well-known for displays of blue-green bioluminescence, but details of its bioluminescent chemistry have been elusive. The glowworm is evolutionarily distant from other bioluminescent creatures studied in detail, including the firefly. We have isolated and characterised the molecular components of the glowworm luciferase-luciferin system using chromatography, mass spectrometry and 1H NMR spectroscopy. The purified luciferase enzyme is in the same protein family as firefly luciferase (31% sequence identity). However, the luciferin substrate of this enzyme is produced from xanthurenic acid and tyrosine, and is entirely different to that of the firefly and known luciferins of other glowing creatures. A candidate luciferin structure is proposed, which needs to be confirmed by chemical synthesis and bioluminescence assays. These findings show that luciferases can evolve independently from the same family of enzymes to produce light using structurally different luciferins.