One of the major obstacles in present plant gene cloning is the lack of integrated genetic/physical maps.
The main objective of this project is to achieve the molecular dissection of the tomato genome into genetically defined sectors of which the physical size is determined. The function of the genes present in these
genetically/physically integrated sectors will be established using saturation mutagenesis and map-based cloning approaches.
In this project leading European tomato groups (7) and companies (2) will combine their expertises in complementary approaches that unite established and novel techniques for tomato genome analysis. More rapid methods for screening and cloning genes of economic relevance will result. This precompetitive genome project will enable future extrapolation of knowledge acquired from the Arabidopsis genome project to important European crop plants. Collaboration between laboratories working on transposon tagging (Jones, Theres, Taylor and van Haaren) and laboratories skilled in map-based cloning techniques (Keygene, Jones, Zabel, Zeneca) will be complemented by laboratories working-on novel strategies for genome analysis based on site-specific recombination and rare cutting restriction enzymes (Gidoni, Murray, van Haaren).
The fundamental of this project is a unique multi-functional T-DNA construct that we have recently constructed. After introduction into the tomato genome this will serve as genetically identifiable landmarks (500), which we will locate on average every 2 Megabases throughout the genome. Specific in-vitro DNA cleavage on these genetic loci enables the construction of physical maps between landmarks, followed by the rapid isolation of molecular markers and the preparation of YAC contigs for each section of the genome. These conventional map-based cloning techniques are combined with novel strategies for screening a genome for new genes based on site-specific recombination reactions, allowing the efficient isolation or conditional knockout of the genes responsible for these traits.
The resulting integrated molecular and physical map of the tomato genome will form the basis for detailed analysis of specific sectors of the tomato genome, which contain genetically or synteny mapped genes of agricultural and industrial importance.
Knowledge and materials produced during this project will be of great importance for many future projects not only in tomato, but also in other important crop plant particularly members of the Solanaceae. The technology developed for tomato during this project can be transferred to other crop plants with even larger genomes that are important for the European Community. Specific aims of this project are the identification and cloning of traits for fungal and nematode resistance (Jones, van Haaren, Zabel and Keygene) and commercially important plant developmental processes (Theres, Taylor, Murray, Gidoni, Zeneca).
Close collaboration between laboratories specialised in tomato genetics and a broad participation of industry will ensure that the scientific knowledge produced during this project will be tested for potential exploitation on the market place.
Funding SchemeCSC - Cost-sharing contracts
6700 AE Wageningen
50250 Bet Dagan
NR4 7UH Norwich
CB2 1QT Cambridge
LE12 5RD Loughborough
6703 HA Wageningen
RG12 6EY Bracknell