Abstract
Insect pests cause billions of dollars in crop losses and there is the ever-present threat of insecticide resistance, pesticide pollution of food and environmental damage. New ways of controlling insect pests are urgently needed. Arginine kinase (AK) is a phosphotransferase, which plays a critical role in cellular energy metabolism in invertebrates. It only presents in invertebrates and may be a suitable chemotherapeutic target in the control of pests. In this study, we cloned and characterized the full-length AK gene from Phyllotreta striolata, one of the most destructive beetle pests worldwide. Furthermore, we constructed a dsRNA targeting AK and used RNAi to control the beetle. The feeding bioassays indicated that minute quantities of dsRNA greatly impaired the beetle's development. Ingestion of dsRNA not only significantly retarded the development and increased the mortality of adults, it also greatly reduced fecundity and fertility, suggesting that RNAi targeting AK is a potential and attractive tool for controlling insect pests.
Highlights
Arginine kinase (ATP: L-arginine phosphotransferase, EC 2.7.3.3, AK) plays a critical role in cellular energy homeostasis in invertebrates by catalyzing a reversible transfer of phosphate from Mg·ATP to arginine, yielding phosphoarginine as follows: Mg·ATP + Arginine Phosphoarginine + Mg·ADP (Newsholme et al, 1978)
AK only exists in the tissues of invertebrates (Lang et al, 1980; Chamberlin, 1997; Kucharski & Maleszka, 1998), and functions as a central regulator of temporal and spatial ATP buffers directly associated with muscle contraction, ATP regeneration and energy transport in cellular energy metabolism (Ellington, 2001)
The open reading frame (ORF) encodes a polypeptide of 356 amino acids with most of the residues considered necessary for AK’s enzymatic function
Summary
Arginine kinase (ATP: L-arginine phosphotransferase, EC 2.7.3.3, AK) plays a critical role in cellular energy homeostasis in invertebrates by catalyzing a reversible transfer of phosphate from Mg·ATP to arginine, yielding phosphoarginine as follows: Mg·ATP + Arginine Phosphoarginine + Mg·ADP (Newsholme et al, 1978). AK plays a critical role in cell energy metabolism in invertebrates and, seems to be an excellent target for RNAi, which would result in an increase in the morbidity and mortality of insects. To determine the adverse effects of dsRNA on P. striolata, adult beetles were grouped and fed a range of dilutions of dsRNA, from 0.05 to 1.60 ng/mL.
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