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The European Wolbachia project: towards novel biotechnological approaches for control of arthropod pests and modification of beneficial arthropod species by endosymbiotic bacteria

   
Project

QLK3-2000-01079

Cell factory area

3.3.4

EU Contribution

2 231 171 Euro

Duration

36 months

Type

Research project

Starting date

01-09-2000

Keywords
arthropod pests
Drosophila melanogaster
European Wolbachia
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ABSTRACT
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Wolbachia may be the most abundant and widespread intracellular symbiont as yet described. This unculturable, maternally inherited bacterium is able to invade and maintain itself in numerous arthropod host species by inducing a variety of reproductive alterations such as induction of parthenogenetic development in certain parasitic wasps, overriding chromosomal sex determination to convert infected genetic males into functional females in crustacea species (feminisation), and most commonly induction of cytoplasmic incompatibility (CI), a form of embryonic lethality in crosses between males and females of different Wolbachia infection status. This proposal aims to identify the bacterial and host genes involved in the induction of these reproductive phenotypes through an integrated genomics, proteomics and post-genomics approach. Identification of these genes will be a major breakthrough towards the goal of using them for applied purposes, for example management of arthropod agricultural pests and disease vectors or improvement of beneficial arthropods.

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OBJECTIVES
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The objectives of the project are:

  • To identify Wolbachia genes that are involved in cytoplasmic incompatibility, parthenogenesis and feminisation, using an integrated genomics, proteomics and post-genomics approach.
  • To identify host genes that are involved in host-Wolbachia interaction, using a post-genomics approach in Wolbachia-infected Drosophila melanogaster.
  • To develop a robust genetic transformation system for Wolbachia, a technology that will enable further functional studies and genetic manipulation of the bacterium for applied purposes.

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DESCRIPTION OF THE WORK
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Candidate Wolbachia genes responsible for the induction of cytoplasmic incompatibility, parthenogenesis and feminisation will be identified initially through comparative genomics and proteomics analysis. For comparative genomics, the genome sequence of three Wolbachia strains, each inducing one of the phenotypes, will be determined through a shot-gun sequencing approach. For proteomics analysis, Wolbachia infected strains and their tetracycline-treated derivatives will be used as sources for protein extracts. Further identification of candidate genes responsible for the induction of cytoplasmic incompatibility will be achieved through comparative microarray-based expression profiling of Wolbachia strains which differ genetically in their ability to induce this phenotype.

Candidate host genes involved in the host-Wolbachia interaction will be identified by a combination of comparative microarray-based expression profiling and proteomics using infected and uninfected Drosophila strains.

An efficient genetic transformation system for Wolbachia will be developed using either homologous recombination or transposition based on Tc1/mariner transposable element vectors.

Involvement of the candidate Wolbachia genes in inducing the phenotypes will be tested by a gene knock-out approach. Candidate genes for the induction of cytoplasmic incompatibility will also be tested by gene complementation of Wolbachia mutants. In the possible absence of a Wolbachia transformation system, Wolbachia cytoplasmic incompatibility genes will be tested by using available germ-line transformation technology for Drosophila melanogaster to express these genes in Drosophila melanogaster tissues.

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DELIVERABLES
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  • The complete and annotated genome sequence of three Wolbachia strains, each responsible for the induction of cytoplasmic incompatibility, parthenogenesis and feminisation.
  • A genetic transformation system for Wolbachia.
  • Identified and characterised Wolbachia genes involved in the induction of the three phenotypes.
  • Identified and characterised host (Drosophila melanogaster) genes.

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CONSORTIUM
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COORDINATOR
  Dr. Kostas Bourtzis
IMMB-FORTH
PO Box 1527
Heraklion 71110, Crete, Greece
Tel: +30 81394541
Fax: +30 81391101
bourtzis@imbb.forth.gr

Prof. Charalambos Savakis
IMMB-FORTH
PO Box 1527
Heraklion 71110, Crete, Greece
Tel: +30 81391114
Fax: +30 81391101
savakis@imbb.forth.gr

Dr. Christos Ouzounis
IMMB-FORTH
PO Box 1527
Heraklion 71110, Crete, Greece
Tel: +30 81391100
Fax: +30 81391101
ouzounis@imbb.forth.gr

PARTNERS
  Assoc.Prof. Siv Andersson
Department of Molecular Evolution
Uppsala University
BMC, BOX 590, Husargatan 3
75124 Uppsala, Sweden
Tel: +46 184714379
Fax: +46 18557723
Siv.Andersson@molbio.uu.se

Prof. Charles Kurland
Department of Molecular Evolution
Uppsala University
BMC, BOX 590, Husargatan 3
75124 Uppsala, Sweden
Tel: +46 184714379
Fax: +46 18557723
chuck@alpha2.bmc.uu.se

Prof. Roger Garrett
Institute of Molecular Biology
University of Copenhagen
Soelvgade 83H
1307K Copenhagen, Denmark
Tel: +45 35322010
Fax: +45 35324220
garrett@mermaid.molbio.ku.dk

Dr. Henk Braig
School of Biological Sciences
University of Wales Bangor
Deiniol Road
LL57 2UW Bangor, United Kingdom
Tel: +44 1248382354
Fax: +44 1248370731
h.braig@bangor.ac.uk

Prof. Michael Ashburner
Department of Genetics
University of Cambridge
Downing Street
CB2 3EH Cambridge, United Kingdom
Tel: +44 1223333969
Fax: +44 1248333992
ma11@gen.cam.ac.uk

Dr. Richard Stouthamer
Department of Plant Sciences
Wageningen University
PO Box 8031, Binnenhaven 7
6700EH Wageningen, The Netherlands
Tel: +31 317482244
Fax: +31 317484821
Richard.Stouthamer@users.ento.wau.nl

Dr. Gilbert Martin
UMR CNRS 6556
Universite de Poitiers
40 Avenue du Recteur Pineau
86022 Poitiers, France
Tel: +33 549454822
Fax: +33 549453050
Gilbert.martin@campus.univ-poitiers.fr

Ms Artemis-Georgia Hatzigeorgiou
Synaptic Ltd
Science and Technological Park of Crete
PO Box 1447
Voutes, Heraklion 711 10, Crete, Greece
Tel: +30 81391867
Fax: +30 81391906
artemis@stepc.gr
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