Transcriptome analysis of the synganglion from the honey bee mite, Varroa destructor and RNAi knockdown of neural peptide targets

Ewan M. Campbell, Giles E. Budge, Max Watkins, Alan S. Bowman

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Abstract

Varroa mites (Varroa destructor) and the viruses that they transmit are one of the major contributing factors to the global honey bee crisis. Gene products within the nervous system are the targets of all the insecticides currently used to control Varroa but there is a paucity of transcriptomic data available for Varroa neural tissues. A cDNA library from the synganglia (“brains”) of adult female Varroa was constructed and 600 ESTs sequenced and analysed revealing several current and potential druggable targets. Contigs coding for the deformed wing virus (DWV) variants V.destructor virus-1 (VDV-1) and the recombinant (VDV-1DVD) were present in the synganglion library. Negative-sense RNA-specific PCR indicated that VDV-1 replicates in the Varroa synganglion and all other tissues tested, but we could not detect DWV replicating in any Varroa tissue. Two neuropeptides were identified in the synganlion EST library: a B-type allatostatin and a member of the crustacean hyperglycaemic hormone (CHH) superfamily. Knockdown of the allatostatin or the CHH-like gene by double-stranded RNA-interference (dsRNAi) resulted in 85% and 55% mortality, respectively, of Varroa. Here, we present the first transcriptomic survey in Varroa and demonstrate that neural genes can be targeted by dsRNAi either for genetic validation of putative targets during drug discovery programmes or as a potential control measure in itself.
Original languageEnglish
Pages (from-to)116-126
Number of pages11
JournalInsect Biochemistry and Molecular Biology
Volume70
Early online date23 Dec 2015
DOIs
Publication statusPublished - Mar 2016

Fingerprint

Varroidae
Acarapis woodi
Varroa destructor
Varroa
Honey
Mites
Bees
Gene Expression Profiling
RNA Interference
transcriptomics
Viruses
peptides
Peptides
Double-Stranded RNA
Genes
Expressed Sequence Tags
Deformed wing virus
Tissue
allatostatin
double-stranded RNA

Keywords

  • Varroa destructor
  • Apis mellifera
  • synganglion
  • deformed wing virus
  • allatostatin
  • crustacean hyperglaecemic hormone
  • gene knockdown

Cite this

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title = "Transcriptome analysis of the synganglion from the honey bee mite, Varroa destructor and RNAi knockdown of neural peptide targets",
abstract = "Varroa mites (Varroa destructor) and the viruses that they transmit are one of the major contributing factors to the global honey bee crisis. Gene products within the nervous system are the targets of all the insecticides currently used to control Varroa but there is a paucity of transcriptomic data available for Varroa neural tissues. A cDNA library from the synganglia (“brains”) of adult female Varroa was constructed and 600 ESTs sequenced and analysed revealing several current and potential druggable targets. Contigs coding for the deformed wing virus (DWV) variants V.destructor virus-1 (VDV-1) and the recombinant (VDV-1DVD) were present in the synganglion library. Negative-sense RNA-specific PCR indicated that VDV-1 replicates in the Varroa synganglion and all other tissues tested, but we could not detect DWV replicating in any Varroa tissue. Two neuropeptides were identified in the synganlion EST library: a B-type allatostatin and a member of the crustacean hyperglycaemic hormone (CHH) superfamily. Knockdown of the allatostatin or the CHH-like gene by double-stranded RNA-interference (dsRNAi) resulted in 85{\%} and 55{\%} mortality, respectively, of Varroa. Here, we present the first transcriptomic survey in Varroa and demonstrate that neural genes can be targeted by dsRNAi either for genetic validation of putative targets during drug discovery programmes or as a potential control measure in itself.",
keywords = "Varroa destructor, Apis mellifera, synganglion, deformed wing virus, allatostatin, crustacean hyperglaecemic hormone, gene knockdown",
author = "Campbell, {Ewan M.} and Budge, {Giles E.} and Max Watkins and Bowman, {Alan S.}",
note = "Acknowledgements This work was funded by BBSRC-LINK grant # BB/J01009X/1 and Vita Europe Ltd. We are grateful to the Scottish Beekeepers Association, especially Mr Phil McAnespie in supporting this work at its inception. We acknowledge partial funding from a Genesis Faraday SPARK Award, part of a Scottish Government SEEKIT project for the early part of this work. We are grateful to Prof David Evans for his advice on Varroa destructor viruses.",
year = "2016",
month = "3",
doi = "10.1016/j.ibmb.2015.12.007",
language = "English",
volume = "70",
pages = "116--126",
journal = "Insect Biochemistry and Molecular Biology",
issn = "0965-1748",
publisher = "Elsevier Limited",

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T1 - Transcriptome analysis of the synganglion from the honey bee mite, Varroa destructor and RNAi knockdown of neural peptide targets

AU - Campbell, Ewan M.

AU - Budge, Giles E.

AU - Watkins, Max

AU - Bowman, Alan S.

N1 - Acknowledgements This work was funded by BBSRC-LINK grant # BB/J01009X/1 and Vita Europe Ltd. We are grateful to the Scottish Beekeepers Association, especially Mr Phil McAnespie in supporting this work at its inception. We acknowledge partial funding from a Genesis Faraday SPARK Award, part of a Scottish Government SEEKIT project for the early part of this work. We are grateful to Prof David Evans for his advice on Varroa destructor viruses.

PY - 2016/3

Y1 - 2016/3

N2 - Varroa mites (Varroa destructor) and the viruses that they transmit are one of the major contributing factors to the global honey bee crisis. Gene products within the nervous system are the targets of all the insecticides currently used to control Varroa but there is a paucity of transcriptomic data available for Varroa neural tissues. A cDNA library from the synganglia (“brains”) of adult female Varroa was constructed and 600 ESTs sequenced and analysed revealing several current and potential druggable targets. Contigs coding for the deformed wing virus (DWV) variants V.destructor virus-1 (VDV-1) and the recombinant (VDV-1DVD) were present in the synganglion library. Negative-sense RNA-specific PCR indicated that VDV-1 replicates in the Varroa synganglion and all other tissues tested, but we could not detect DWV replicating in any Varroa tissue. Two neuropeptides were identified in the synganlion EST library: a B-type allatostatin and a member of the crustacean hyperglycaemic hormone (CHH) superfamily. Knockdown of the allatostatin or the CHH-like gene by double-stranded RNA-interference (dsRNAi) resulted in 85% and 55% mortality, respectively, of Varroa. Here, we present the first transcriptomic survey in Varroa and demonstrate that neural genes can be targeted by dsRNAi either for genetic validation of putative targets during drug discovery programmes or as a potential control measure in itself.

AB - Varroa mites (Varroa destructor) and the viruses that they transmit are one of the major contributing factors to the global honey bee crisis. Gene products within the nervous system are the targets of all the insecticides currently used to control Varroa but there is a paucity of transcriptomic data available for Varroa neural tissues. A cDNA library from the synganglia (“brains”) of adult female Varroa was constructed and 600 ESTs sequenced and analysed revealing several current and potential druggable targets. Contigs coding for the deformed wing virus (DWV) variants V.destructor virus-1 (VDV-1) and the recombinant (VDV-1DVD) were present in the synganglion library. Negative-sense RNA-specific PCR indicated that VDV-1 replicates in the Varroa synganglion and all other tissues tested, but we could not detect DWV replicating in any Varroa tissue. Two neuropeptides were identified in the synganlion EST library: a B-type allatostatin and a member of the crustacean hyperglycaemic hormone (CHH) superfamily. Knockdown of the allatostatin or the CHH-like gene by double-stranded RNA-interference (dsRNAi) resulted in 85% and 55% mortality, respectively, of Varroa. Here, we present the first transcriptomic survey in Varroa and demonstrate that neural genes can be targeted by dsRNAi either for genetic validation of putative targets during drug discovery programmes or as a potential control measure in itself.

KW - Varroa destructor

KW - Apis mellifera

KW - synganglion

KW - deformed wing virus

KW - allatostatin

KW - crustacean hyperglaecemic hormone

KW - gene knockdown

U2 - 10.1016/j.ibmb.2015.12.007

DO - 10.1016/j.ibmb.2015.12.007

M3 - Article

VL - 70

SP - 116

EP - 126

JO - Insect Biochemistry and Molecular Biology

JF - Insect Biochemistry and Molecular Biology

SN - 0965-1748

ER -