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Development and use of Arabidopsis Thaliana as a tool for isolating genes of agronomic importance

Objective

This T project provides a framework that links scientists working on methods for gene identification to scientists dedicated to answering fundamental questions in plant biology using genetics and physiology as their disciplines. This linkage will accelerate the dissemination of techniques for gene search, and their applications to the molecular characterization of genes involved in flowering, seed development and embryogenesis.
Some of the major scientific goals met in the Arabidopsis thaliana programme are as follows. Approximately one third of the genome has been mapped, techniques for rapidly screening yeast artificial chromosome (YAC) libraries have been developed, and a database has been established for collating mapping and genetic data. Useful frequencies of homologous recombination in Arabidopsis have been demonstrated using the reconstruction of a hygromycin resistence gene as a selection for these rare events. These constructs are being made available for testing recombination methods and deoxyribonucleic acid (DNA) delivery systems. Important new mutations affecting abscisic acid action have been isolated, and the effect of these on gene expression, lipid and sugar metabolism, and interactions with other mutants are being investigated. An 11 kb piece of DNA has been identified, using chromosome walking, that complement abi3 mutations. With this gene identified, its sequence will help reveal the mode of action of a key growth regulator in plants. T-DNA tagged loci have been identified that affect the tissue specificity of GUS expression. Several of these effect embryo specific patterns of expression. A gene encoding the APETALATA2 product has been tagged using T-DNA and isolated. This gene, which determines floral morphology together with a small group of other homeotic genes, has been further characterised by sequencing. A walk using YACs of about 1 600kb has covered the co locus, which conditions flowering time. Several recombinants have narrowed down the region containing the gene, and complementation experiments are underway to isolate clones containing the gene. Similar walks are progressing towards the fca, fwa and fve loci. In addition, epistatic interactions between the different classes of mutants are being studied. Systems that drive high frequency excision and methods for selecting reintegration events, in transposon tagging have been developed. Large populations of plants are being screened for mutations.
This is a coordinated proposal from 44 participants in 9 countries that aims to develop techniques for gene isolation from Arabidopsis and use these methods for isolating genes involved in different aspects of plant biology relevant to agricultural development. There are 7 industrial contractors who will contribute directly to the proposal. Refinement of Arabidopsis as a genetic tool will be accomplished by groups for physical mapping, transposon tagging, gene replacement and resource distribution. Surrounding this are 5 groups who propose to concentrate their efforts on floral initiation, plant growth and differentiation, metabolic control, disease resistance, and seed development and stress responses. The proposed groups are based on areas of common technical or biological expertise and aims, and have been coordinated to provide a balance between technological and biological development that will, if implemented, lead to the rapid development of Arabidopsis as a tool for isolating genes of interest to agriculture. This framework has been designed to accommodate further biological areas of agricultural interest. Existing international collaborations Will be strengthened by this framework and will provide an efficient approach to the physical mapping and cloning of the Arabidopsis genome. At the end of the proposed work there will be sufficient data to initiate a programme of ordered sequencing of the entire genome.

Coordinator

JOHN INNES CENTRE
Address
Norwich Research Park, Colney
Norwich
United Kingdom

Participants (19)

Centre National de la Recherche Scientifique
France
Address
Avenue De Villeneuve
66860 Perpignan
Centre for Plant Breeding Research
Netherlands
Address
6,Keijenbergseweg
6700 AA Wageningen
Consejo Superior de Investigaciones Cientòficas
Spain
Address
18-26,Jordi Girona
08034 Barcelona
DANISCO A/S
Denmark
Address
1,Langebrogade 1
1001 Koebenhavn K/copenhaegen
INSTITUTO NACIONAL DE INVESTIGACION Y TECNOLOGIA AGRARIA Y ALIMENTARIA
Spain
Address
Km.7,carretera De La Coruça Km.7
28040 Madrid
Institut für Genbiologische Forschung Berlin GmbH
Germany
Address
Ihnestraße 63
14195 Berlin
KEYGENE NV
Netherlands
Address
90, 90 Agro Business Park
6700 AE Wageningen
LEIDEN UNIVERSITY
Netherlands
Address
64,Wassenaarseweg 64
2333 AL Leiden
Max-Planck-Gesellschaft zur Forderungder Wissenschaften e.V.
Germany
Address
Karl-von-linné-weg 10
50829 Köln
Mogen International NV trading as ZENECA MOGEN
Netherlands
Address
97,Einsteinweg
2333 CB Leiden
Plant Genetics Systems NV
Belgium
Address
22,Jozef Plateaustraat
9000 Gent
SG Seeds BV
Netherlands
Address
62,Westeinde
1600 AA Enkhuizen
SWISS FEDERAL INSTITUTE OF TECHNOLOGY ZUERICH
Switzerland
Address
2,Universitaetstrasse 2
8092 Zurich
THE PROVOST, FELLOWS AND SCHOLARS OF THE COLLEGE OF THE HOLY AND UNDIVIDED TRINITY OF QUEEN ELIZABETH NEAR DUBLIN HEREINAFTER TRINITY COLLEGE DUBLIN
Ireland
Address
Smurfit Institute,trinity College
2 Dublin
Universiteit Gent
Belgium
Address
35,K.l. Ledeganckstraat 35
9000 Gent
University of Leicester
United Kingdom
Address
University Road
LE1 7RH Leicester
University of Nottingham
United Kingdom
Address
University Park
NG7 2RD Nottingham
WAGENINGEN UNIVERSITY
Netherlands
Address
4,Arboretumlaan 4
6703 BD Wageningen
Zeneca Group plc
United Kingdom
Address
Fernhurst
GU27 3JE Haslemere