Bowman, Alan S.

Budge, Giles E.

Campbell, Ewan M.

Parasites & Vectors

English

PubMed

Extent: 10p.Background: The parasitic mite Varroa destructor is considered the major pest of the European honey bee (Apis mellifera) and responsible for declines in honey bee populations worldwide. Exploiting the full potential of gene sequences becoming available for V. destructor requires adaptation of modern molecular biology approaches to this non-model organism. Using a mu-class glutathione S-transferase (VdGST-mu1) as a candidate gene we investigated the feasibility of gene knockdown in V. destructor by double-stranded RNA-interference (dsRNAi). Results: Intra-haemocoelic injection of dsRNA-VdGST-mu1 resulted in 97% reduction in VdGST-mu1 transcript levels 48 h post-injection compared to mites injected with a bolus of irrelevant dsRNA (LacZ). This gene suppression was maintained to, at least, 72 h. Total GST catalytic activity was reduced by 54% in VdGST-mu1 gene knockdown mites demonstrating the knockdown was effective at the translation step as well as the transcription steps. Although near total gene knockdown was achieved by intra-haemocoelic injection, only half of such treated mites survived this traumatic method of dsRNA administration and less invasive methods were assessed. V. destructor immersed overnight in 0.9% NaCl solution containing dsRNA exhibited excellent reduction in VdGST-mu1 transcript levels (87% compared to mites immersed in dsRNA-LacZ). Importantly, mites undergoing the immersion approach had greatly improved survival (75-80%) over 72 h, approaching that of mites not undergoing any treatment. Conclusions: Our findings on V. destructor are the first report of gene knockdown in any mite species and demonstrate that the small size of such organisms is not a major impediment to applying gene knockdown approaches to the study of such parasitic pests. The immersion in dsRNA solution method provides an easy, inexpensive, relatively high throughput method of gene silencing suitable for studies in V. destructor, other small mites and immature stages of ticks.Ewan M Campbell, Giles E Budge, Alan S Bowma

Campbell, E.

Budge, G.

Bowman, A.

Adelaide Research & Scholarship

Gene-knockdown in the honey bee mite Varroa destructor by a non-invasive approach: studies on a glutathione S-transferase

Aberdeen University Research

Acaricide (pyrethroid) resistance in Varroa destructor.

AE: A water-specific aquaporin involved in aphid osmoregulation. Insect Biochem Mol Biol

AS: RNA-interference methods for geneknockdown in the sea louse, Lepeophtheirus salmonis: Studies on a putative prostaglandin E synthase. Parasitology

Biochemical markers linked to abamectin resistance in Tetranychus urticae (Acari: Tetranychidae). Pestic Biochem Physiol

Bowman AS: Role of an aquaporin in the sheep tick Ixodes ricinus: Assessment as a potential control target.

CA: Structure, function and evolution of glutathione transferases: Implications for classification of non-mammalian members of an ancient enzyme superfamily.

EF: RNA interference for the study and genetic manipulation of ticks. Trends Parasitol

Fluorometric microplate assay to measure glutathione S-transferase activity in insects and mites using monochlorobimane. Anal Biochem

GJ: Jalview Version 2-A multiple sequence alignment editor and analysis workbench. Bioinformatics

Higgins DG: Clustal W and Clustal X version 2.0. Bioinformatics

Ingested double-stranded RNAs can act as species-specific insecticides. Insect Biochem Mol Biol

JD: Genomic survey of the ectoparasitic mite Varroa destructor, a major pest of the honey bee Apis mellifera.

JS: Increased transcription of glutathione S-transferases in acaricide exposed scabies mites. Parasites and Vectors

Krüppel homolog 1, an early juvenile hormone-response gene downstream of Methoprene-tolerant, mediates its anti-metamorphic action in the red flour beetle Tribolium castaneum. Dev Biol

LR: Prolonged gene knockdown in the tsetse fly Glossina by feeding double stranded RNA. Insect Mol Biol

MatheyPrevot B: Evidence of off-target effects associated with long dsRNAs in Drosophila melanogaster cell-based assays. Nature Methods

Mattsson JG: Effect of acaricides on the activity of glutathione transferases from the parasitic mite Sarcoptes scabiei. Parasitology

MD: A historical review of managed honey bee populations in Europe and the United States and the factors that may affect them.

Parasitic mites of honey bees: Life history, implications, and impact. Annu Rev Entomol.

Propetamphos resistance in Rhipicephalus bursa (Acari, Ixodidae). Vet Parasitol

Pyrethroid insecticide resistance in Rhipicephalus bursa (Acari, Ixodidae). Pestic Biochem Physiol

S: MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol

The effect of insecticide synergists on the response of scabies mites to pyrethroid acaricides. PLoS Neglected Tropical Diseases 2009, 3(1):e354, Epub

The honey bee colony: Life history. The hive and the honey bee Hamilton Illinois:

The role of varroa and viral pathogens in the collapse of honey bee colonies: A modelling approach.

Time-activity budgets and space structuring by the different life stages of Varroa jacobsoni in capped brood of the honey bee, Apis mellifera.

Tscharntke T: Importance of pollinators in changing landscapes for world crops.

Vincenta DL: Comparative morphology of the peritremes of Varroa and Euvarroa (Varroidae), parasites of honey bees (Apidae).

Gene-knockdown in the honey bee mite Varroa destructor by a non-invasive approach : studies on a glutathione S-transferase

Ewan M Campbell

Giles E Budge

Alan S Bowman

Springer - Publisher Connector

Abstract Background The parasitic mite Varroa destructor is considered the major pest of the European honey bee (Apis mellifera) and responsible for declines in honey bee populations worldwide. Exploiting the full potential of gene sequences becoming available for V. destructor requires adaptation of modern molecular biology approaches to this non-model organism. Using a mu-class glutathione S-transferase (VdGST-mu1) as a candidate gene we investigated the feasibility of gene knockdown in V. destructor by double-stranded RNA-interference (dsRNAi). Results Intra-haemocoelic injection of dsRNA-VdGST-mu1 resulted in 97% reduction in VdGST-mu1 transcript levels 48 h post-injection compared to mites injected with a bolus of irrelevant dsRNA (LacZ). This gene suppression was maintained to, at least, 72 h. Total GST catalytic activity was reduced by 54% in VdGST-mu1 gene knockdown mites demonstrating the knockdown was effective at the translation step as well as the transcription steps. Although near total gene knockdown was achieved by intra-haemocoelic injection, only half of such treated mites survived this traumatic method of dsRNA administration and less invasive methods were assessed. V. destructor immersed overnight in 0.9% NaCl solution containing dsRNA exhibited excellent reduction in VdGST-mu1 transcript levels (87% compared to mites immersed in dsRNA-LacZ). Importantly, mites undergoing the immersion approach had greatly improved survival (75-80%) over 72 h, approaching that of mites not undergoing any treatment. Conclusions Our findings on V. destructor are the first report of gene knockdown in any mite species and demonstrate that the small size of such organisms is not a major impediment to applying gene knockdown approaches to the study of such parasitic pests. The immersion in dsRNA solution method provides an easy, inexpensive, relatively high throughput method of gene silencing suitable for studies in V. destructor, other small mites and immature stages of ticks.</p

Campbell Ewan M

Budge Giles E

Bowman Alan S

Directory of Open Access Journals

Gene-knockdown in the honey bee mite <it>Varroa destructor </it>by a non-invasive approach: studies on a glutathione S-transferase

Abstract
          
            Background
            The parasitic mite Varroa destructor is considered the major pest of the European honey bee (Apis mellifera) and responsible for declines in honey bee populations worldwide. Exploiting the full potential of gene sequences becoming available for V. destructor requires adaptation of modern molecular biology approaches to this non-model organism. Using a mu-class glutathione S-transferase (Vd GST-mu1) as a candidate gene we investigated the feasibility of gene knockdown in V. destructor by double-stranded RNA-interference (dsRNAi).
          
          
            Results
            Intra-haemocoelic injection of dsRNA-Vd GST-mu1 resulted in 97% reduction in Vd GST-mu1 transcript levels 48 h post-injection compared to mites injected with a bolus of irrelevant dsRNA (LacZ). This gene suppression was maintained to, at least, 72 h. Total GST catalytic activity was reduced by 54% in Vd GST-mu1 gene knockdown mites demonstrating the knockdown was effective at the translation step as well as the transcription steps. Although near total gene knockdown was achieved by intra-haemocoelic injection, only half of such treated mites survived this traumatic method of dsRNA administration and less invasive methods were assessed. V. destructor immersed overnight in 0.9% NaCl solution containing dsRNA exhibited excellent reduction in Vd GST-mu1 transcript levels (87% compared to mites immersed in dsRNA-LacZ). Importantly, mites undergoing the immersion approach had greatly improved survival (75-80%) over 72 h, approaching that of mites not undergoing any treatment.
          
          
            Conclusions
            Our findings on V. destructor are the first report of gene knockdown in any mite species and demonstrate that the small size of such organisms is not a major impediment to applying gene knockdown approaches to the study of such parasitic pests. The immersion in dsRNA solution method provides an easy, inexpensive, relatively high throughput method of gene silencing suitable for studies in V. destructor, other small mites and immature stages of ticks.
          </jats:sec

Crossref

https://aura.abdn.ac.uk/bitstream/2164/859/1/mite%20geneknockdown.pdf

Gene-knockdown in the honey bee mite Varroa destructor by a non-invasive approach : studies on a glutathione S-transferase

Abstract

Similar works

Full text

Available Versions

Adelaide Research & Scholarship

Aberdeen University Research

Springer - Publisher Connector

Directory of Open Access Journals

Crossref