Work Package 3 (ESR 4)

Nasonia MUTAGENESIS

General details

NetherlandsUKLocation
University of Leicester, UK
University of Groningen, NL

Supervisors
Dr. E. Tauber
Prof L. Beukeboom
Dr L. van de Zande

ESR 4
Marcela Buřičová

Secondments

The ESR will be funded half the time in Leicester and half the time in Groningen. The
major secondment is between these two institutions and is inbuilt into this arrangement.

Objectives

  • Identify genes underlying photoperiodic timing in Nasonia using chemical mutagenesis.
  • Characterise the expression and function of photoperiodic genes identified above.

Methods

  •  Isolate photoperiodic mutants induced by EMS mutagenesis.
  •  Identify the DNA sequence changes underlying the mutations.
  •  Characterise the expression of photoperiodic genes and carry functional assays.

Description

Mutagenesis screen for photoperiodic timing genes in Nasonia
Seasonal timing is a key process for the successful survival and reproduction of many organisms, especially in temperate regions. In most cases of seasonal timing, light is the primary cue and the seasonal change in day-length is monitored by a so-called photoperiodic clock. In insects, this clock regulates entry into a specialised state of dormancy, termed diapause. Diapause is an overwintering strategy used by virtually all insects, and is critical in regulating their phenology, distribution and abundance. Despite intensive study of the photoperiodic clock at the phenotypic level over the last 80 years there still remains something of a ‘black box’ in terms of the underlying molecular mechanisms. Here, we going use Nasonia vitripennis, an emerging model organism (whose genome has been recently sequenced) that shows a strong photoperiodic diapause response: short photoperiods experienced by females induce larval diapause.
We will use chemical mutagenesis to identify genetic variants that affect circadian and photoperiodic timing, by mating chemically treated males to genetically marked females. F1 females will be kept in short days, and the progeny of each female will be tested for reduced levels of diapause. Separately, F1 males will be tested for their circadian behaviour, and putative mutants with aberrant rhythmicity will be isolated (males are haploid, so recessive mutations are immediately apparent). The second phase of the project, using linkage analysis to map the mutations, will be carried out in Groningen. By taking advantage of the strains that have been recently sequenced in Groningen, and recent SNP mapping protocols developed for Drosophila, mapping can be completed within ~6 months.  The third phase of the project will be in Leicester, and will characterise the various mutations by dissecting their role in circadian and photoperiodic control, studying the expression of the corresponding  genes by in-situ hybridisation and their functional analysis by dsRNAi knockdown.

Results and milestones

  1.  Submit a first year report by month 12.
  2.  Complete mutant mapping by month 30.
  3.  Submit a publication by month 48

Synergies, Risks & Exploitation

University of Leicester takes advantage of the strains that have been recently sequenced and are available in Groningen, together with novel SNP methodology developed for Drosophila. The genes identified can serve as targets for developing genetically modified insects for pest control or by generating superior parasitic wasps.

See more Work Packages in Research Area A