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Target Verification of Allyl Isothiocyanate on the Core Subunits of Cytochrome C Oxidase in Sitophilus zeamais by RNAi.

BACKGROUND
Allyl isothiocyanate (AITC) is a volatile organic compound with a potent insecticidal activity to the stored-grain pest Sitophilus zeamais Motschulsky, which severely damages grain storage and container transport worldwide. Our previous study showed that mitochondrial complex IV was the primary target of AITC in adult S. zeamais. To further verify the targets of AITC, we employed RNA interference (RNAi) by using double-stranded RNA (dsRNA) to knockdown three core subunits of Cytochrome C Oxidase (COX) I, II and III in 18-day-old larvae prior to their exposure to AITC to detect susceptibility changes.


RESULTS
The susceptibility of dsRNACOX I and II injection treatments to AITC significantly increased at 72 h while the mortality reached up to 85.56% and 67.78%, respectively, and dsRNACOX I and dsRNACOX II injection showed the same subcellular structural characteristics appearing vacuolization and vague mitochondrial cristae and decrease of COX activity as AITC fumigation treatment, suggesting the potential of COX I and COX II as the targets of AITC. High mortality reaching up to 75.55%, 71.88% and 82.22% and the phenotype that larvae turning from milky white to dark brown in the thorax and death eventually were showed after dsRNACOX I, II and III injection.


CONCLUSION
COX I and II were elucidated as the potential targets of AITC and dsRNACOX I, II and III have the potential to be developed into nucleic acid pesticides for their robust lethal effects and are worth pursuing for improving AITC fumigation activity in S. zeamais control. This article is protected by copyright. All rights reserved.