Molecular, biochemical, and toxicological characterization of two acetylcholinesterases from the Western flower thrips, Frankliniella occidentalis

Jong Hyeok Lee; In Hong Jeong; Dong Woon Lee; Si Hyeock Lee; Kyungjae Andrew Yoon

doi:10.1016/j.pestbp.2023.105629

Molecular, biochemical, and toxicological characterization of two acetylcholinesterases from the Western flower thrips, Frankliniella occidentalis

Pestic Biochem Physiol. 2023 Nov:196:105629. doi: 10.1016/j.pestbp.2023.105629. Epub 2023 Sep 22.

Authors

Jong Hyeok Lee¹, In Hong Jeong², Dong Woon Lee³, Si Hyeock Lee⁴, Kyungjae Andrew Yoon⁵

Affiliations

¹ Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea.
² Division of Crop Protection, National Institute of Agricultural Science, Rural Development Administration, Republic of Korea.
³ Department of Ecological Science, Kyungpook National University, Sangju 37224, Republic of Korea.
⁴ Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: shlee22@snu.ac.kr.
⁵ Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: kongbob89@snu.ac.kr.

PMID: 37945233
DOI: 10.1016/j.pestbp.2023.105629

Abstract

We investigated the molecular and biochemical properties of two acetylcholinesterases (FoAChE1 and FoAChE2) from the Western flower thrips, Frankliniella occidentalis. Polyacrylamide gel electrophoresis and western blotting confirmed the membrane-anchored nature of both FoAChE1 and FoAChE2, which was further supported by hydrophobicity and glycophosphatidylinositol anchor predictions. High expression levels of both enzymes were observed in the head, indicating their predominant distribution in neuronal tissues. FoAChE1 exhibited significantly higher expression levels in all examined tissues compared to FoAChE2, suggesting its major role as a synaptic enzyme. Nonetheless, both recombinant enzymes displayed robust catalytic activity toward acetylthiocholine iodide, and FoAChE1 demonstrated nearly identical catalytic efficiency compared to FoAChE2. FoAChE1 exhibited slightly lower sensitivities to the cholinesterase inhibitors tested, including organophosphates (OPs) and carbamates (CBs), compared to FoAChE2. Field populations of F. occidentalis exhibited polymorphism of alanine vs. serine at position 197 of FoAChE1 within the conserved oxyanion hole. Contrary to common belief, however, functional analysis using recombinant enzymes revealed that neither A197 nor S197 residue was associated with FoAChE1 insensitivity to OPs and CBs. FoAChE2 did not exhibit any polymorphic amino acid substitutions at the positions known to be associated with resistance. Due to the absence of apparent resistance-associated mutations in field populations of F. occidentalis, the judicious use of some OPs or CBs can be suggested for controlling the highly resistant populations to other insecticides. Overall, our findings highlight the significance of both FoAChE1 and FoAChE2 as targets for toxicity assessment, while the specific contribution of each enzyme to toxicity remains unclear.

Keywords: Acetylcholinesterase; Frankliniella occidentalis; Functional expression; Resistance; Western flower thrips.

MeSH terms

Acetylcholinesterase / genetics
Animals
Carbamates
Flowers
Insecticides* / toxicity
Thysanoptera*

Substances

Acetylcholinesterase
Insecticides
Carbamates