Oilseed rape variety Express 617	Oilseed rape and Arabidopsis are close relatives. For this reason oilseed rape represents a promising species for the transfer of advances in plant genome research with the goal of commercial crop improvement.
	EMS-mutagenised M₁-plants
In order to develop new genetic diversity useful for direct integration into the actual breeding programs, TILLING is a valuable tool. Therefore the winter oilseed rape variety Brassica napus Express 617 is used to establish the TILLING population. About 7500 M₂-plants comprise the current population.
Open pollination and selfing of M₂-plants	Seed mutagenesis, plant cultivation, sampling, selfing, seed harvesting and phenotypic scoring is in the breeders hand, while the technical improvement of DNA-extraction, gene specific primer design, reverse mutant screening on different platforms (Li-Cor- or ABIXL- DNA analyzer) and molecular characterization of the mutants belong to the academics work package.
For the identification of mutantes in order to test gene function we first priorise on the analysis of key target traits that are of great importance for the participating breeders.

Results

The success of seed mutagenesis with EMS is obvious through scoring the morphological diversity of M₂-plants.

Sampling of almost equal amounts of leave tissue lead to uniform DNA amounts

gDNA-extraction in 96-well format

Screening for allel specific primers in an allopolyploid organism

The specificity of target gene amplification depends on the availability of sequence information.

Prove of concept as well as first mutant screenings using different target genes are performed. The identification of mutant pools is performed on to platforms (Li-Cor und ABI) in parallel.

Fragment analysis on the capillary DNA sequencer

Project partners

	Dieter Stelling, Zeljko Micic, Inga Kottmann
	Milena Ouzunova, Frank Breuer, Frank Weber
	Gunhild Leckband, Felix Dreyer, Amine Abbadi
	Bernd Weisshaar, Daniela Holtgräwe, Thomas Rosleff Sörensen, Stefanie Garbe, Prisca Viehöver
	Christian Jung, Hans Harloff, Susanne Lemcke, Gislind Bräcker

Last Updated ( Monday, 16 February 2009 )