ACE: an efficient and sensitive tool to detect insecticide resistance-associated mutations in insect acetylcholinesterase from RNA-Seq data

Abstract

BackgroundInsecticide resistance is a substantial problem in controlling agricultural and medical pests. Detecting target site mutations is crucial to manage insecticide resistance. Though PCR-based methods have been widely used in this field, they are time-consuming and inefficient, and typically have a high false positive rate. Acetylcholinesterases (Ace) is the neural target of the widely used organophosphate (OP) and carbamate insecticides. However, there is not any software available to detect insecticide resistance associated mutations in RNA-Seq data at present.ResultsA computational pipeline ACE was developed to detect resistance mutations of ace in insect RNA-Seq data. Known ace resistance mutations were collected and used as a reference. We constructed a Web server for ACE, and the standalone software in both Linux and Windows versions is available for download. ACE was used to analyse 971 RNA-Seq data from 136 studies in 7 insect pests. The mutation frequency of each RNA-Seq dataset was calculated. The results indicated that the resistance frequency was 30%–44% in an eastern Ugandan Anopheles population, thus suggesting this resistance-conferring mutation has reached high frequency in these mosquitoes in Uganda. Analyses of RNA-Seq data from the diamondback moth Plutella xylostella indicated that the G227A mutation was positively related with resistance levels to organophosphate or carbamate insecticides. The wasp Nasonia vitripennis had a low frequency of resistant reads (<5%), but the agricultural pests Chilo suppressalis and Bemisia tabaci had a high resistance frequency. All ace reads in the 30 B. tabaci RNA-Seq data were resistant reads, suggesting that insecticide resistance has spread to very high frequency in B. tabaci. ConclusionsTo the best of our knowledge, the ACE pipeline is the first tool to detect resistance mutations from RNA-Seq data, and it facilitates the full utilization of large-scale genetic data obtained by using next-generation sequencing.Electronic supplementary materialThe online version of this article (doi:10.1186/s12859-017-1741-6) contains supplementary material, which is available to authorized users.

Development of molecular assays to detect target-site mechanisms associated with insecticide resistance in malaria vectors from Latin America

“…darlingi as compared with An. gambiae; these substitutions have not been previously reported before in any insect or associated with insecticide resistance [31].…”

Laboratory and field evaluation of the aphidicidal activity of moso bamboo (Phyllostachys pubescens) leaf extract and identification of the active components

“…These compounds were isolated from the petroleum ether component of the bamboo leaf extract and identified with NMR and LC-MS. These flavonoids were used to assay the mustard aphid, and the LC 50 (Table 2), and the isoorientin showed the best aphidicidal activity which was greater than that of the positive control in 24 h, matrine, which was 376.06 and 190.51 mg L −1 , respectively. Then, the flavonoids were identified as the main active aphidicidal components in moso bamboo, and isoorientin was determined to be the most active compound.…”

“…The study of the insecticidal mechanism is an important part of the utilization of a botanical insecticide, and the variety of protective enzymes in insects have become priority targets of botanical insecticide . Acetylcholinesterase (AChE) is an important hydrolase in the insect nervous system that can stop nerve impulses by catalysing the hydrolysis of neurotransmitter choline . Superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) are important antioxidant enzymes in insects because these enzymes can remove free superoxide radicals and maintain a state of dynamic equilibrium, with the free radicals being maintained at a low level to prevent free radical poisoning .…”

“…48,49 Acetylcholinesterase (AChE) is an important hydrolase in the insect nervous system that can stop nerve impulses by catalysing the hydrolysis of neurotransmitter choline. 50,51 Superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) are important antioxidant enzymes in insects because these enzymes can remove free superoxide radicals and maintain a state of dynamic equilibrium, with the free radicals being maintained at a low level to prevent free radical poisoning. [52][53][54] When this balance is destroyed, the insect may be damaged.…”

Caligus rogercresseyi acetylcholinesterase types and variants: a potential marker for organophosphate resistance

“…Mutations in the gene coding for the enzyme acetylcholinesterase (AChE) have been reported as the main cause of resistance towards organophosphates in arthropods, followed by metabolic and behavioural mechanisms (discussed in [ 21 , 24 – 28 ]). AChE is present in the cholinergic synapses of the nervous system of arthropods and in the neuromuscular junctions of vertebrates [ 29 ].…”