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Development of ryegrass allele-specific (GRASP) markers for sustainable grassland improvement

The purpose of this project is to fill the current gap that exists between knowledge on plant gene functions and their commercial application. Bridging this gap, by using the allelic selection tools to be developed within this project, will lead to efficient plant breeding of highly improved, non-GMO crop plants. Once developed, the SNP markers will be made available to commercial breeding companies and public institutions on a European-wide basis for 'direct selection' strategies at the DNA level, to explore germplasm collections, and to describe varieties for intellectual property right protection. Established techniques and approaches can be easily expanded to additional candidate genes. Strategies for routine and cost-effective monitoring will be provided, and for the exploitation of allelic diversity for improved breeding strategies as an element of sustainable agricultural practices.

The overall objective of this project is to establish and validate a framework for the development of allele-specific, gene-derived single nucleotide polymorphism (SNP) markers, associated with agronomically important traits in Lolium, which are relevant to environmental and nutritive value needs.
A set of 20 Lolium genotypes, highly differing for the traits of interest, has already been assembled. The genetic distance among them will be determined using molecular markers, and candidate genes for forage quality and for traits improving environmental sustainability in Lolium will be isolated. Two approaches will be used to determine which candidate genes will be studied:
1) sequence homology
2) differential gene expression.
In parallel, a BAC library will be produced, which will be used for full-length gene isolation. In the second year, a standard protocol will be employed to characterise the 100 candidate genes isolated within this project, including genetic mapping. Gene-specific sequence variation will be studied on the set of 20 Lolium genotypes and all molecular (sequence, map) data will be made available through a database for development of SNP markers used to study allelic variation.
In the following phase, trait specific selections will be carried out over at least two generations using the set of 20 genotypes as starting materials. Changes in allele frequencies associated with divergent selection will be used for validation of candidate genes. The last part of the project is dedicated to the technology transfer including:
1) the development of SNP marker tool kits for allele discrimination in grass breeding
2) making BAC and cDNA / ESTs libraries available
3) selecting breeding populations for specific traits and evaluating them at the level of several relevant genes
4) the development of new breeding strategies implementing allele specific SNP markers developed in this project.

Progress to Date
Within the second year of the project GRASP the major focus was on the establishment of plant materials, molecular resources, and the isolation of L. perenne genes. Relevant resources are now available such as a BAC library, a cDNA microarray for expression profiling, and populations for selection experiments. More than 100 candidate gene sequences for the traits of interest have been isolated, and the most relevant for GRASP are currently selected based on, e.g., expression profiling experiments. An integrated Lolium gene database with access system has been implemented. Mapping and full-length gene isolation using different mapping populations and the BAC library is ongoing, allele sequencing has been initiated. Furthermore, the first selection experiments have been completed and the respective next generations are currently produced for the next cycle of selection.


Scientist responsible for the project

Research Centre Flakkebjerg, Forsoegsvej 1
4200 Slagelse
Denmark - DK

Phone: +45 58113484
Fax: +45 58113301


Project ID QLRT-2001-00862
Area 5.1.1
Start date 01 October 2002
Duration (months) 48
Total cost 5 926 508 €
Total EC contribution   3 237 1745 €
Status Ongoing
Web address of the project

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