THE METHODS DEVELOPED HERE WILL PROVIDE INFORMATION FOR FUTURE ANALYSIS OF THE UNKNOWN PROCESSES BRIDGING THE GAP BETWEEN CELL SURFACE/MEMBRANE EVENTS AND INDUCTION/TRANSLATION EVENTS AT THE GENOMIC LEVEL.
FURTHERMORE THE PLANT DISEASES CAUSED BY ATTACK OF MICROORGANISMS ARE INITIATED BY AN INTERACTION BETWEEN THE PLANT CELL WALL/MEMBRANE AND THE MICROORGANISM (RECOGNITION) SO THAT IT CAN BE EXPECTED THAT RESISTANCE OR SUSCEPTIBILITY IS A MATTER OF PLANT CELL WALL/MEMBRANE BEHAVIOUR.
FROM THIS IT IS LIKELY TO ANTICIPATE THAT TWO OF THE MOST URGENT PROBLEMS TO BE SOLVED - REGENERATION FROM ISOLATED CELLS AND RESISTANCE TO PATHOGENS - ARE BOTH RELATED TO AND CAN BE ANALYZED THROUGH EVENTS TAKING PLACE IN THE CELL WALL OR MEMBRANE.
THE MAIN CONTRACTOR (DDS) HAS A MAIN BREEDING STATION (MARIBO) IN DANMARK WITH BRANCHES IN ENGLAND, FRANCE, ITALY, SPAIN AND AUSTRIA WHICH WILL BE ABLE TO USE THE NEW METHODS IMMEDIATELY. THE CLOSE RELATION TO THESE STATIONS ALSO MAKES POSSIBLE THE USE OF RELEVANT VARIETIES SO THAT BENEFITS MAY BE SEEN ALSO ON A SHORTER TERM.
The kit contains affinity chromatography columns based on monoclonal antibodies for the purification of the plant hormone zeatin. The hormone is then quantitatively assayed through an enzyme-linked-immunosorbent-assay, based on the principle of competitive binding. Femtomole amounts of zeatin could be detected in plant extracts using this kit.
To apply molecular biology and transgenic plants to plant breeding and industrial scale production, and efficient regeneration of entire plants from cell and tissue culture is needed. To do so, the decisive steps in those processes of plant cell differentiation leading to regeneration of plants, somatic embryogenesis must be analysed. This developmental pathway was studied in a model (carrot) and a crop (sugar beet) species to provide new understanding and molecular tools for use in nonresponding species.
A high frequency plant regeneration system from sugar beet suspensions was established and compared to the carrot model system; the carrot system is much better and does not seem to involve somatic embryogenesis regularly. Regenerative capacity was correlated with biochemical markers (eg, isoperoxidases), endogenous cytokinins) and the level of deoxyribonucleic acid (DNA) methylation. A novel technology for ring coupling between plant hormone molecules and large carrier proteins (eg bovine serum albumin (BSA) resulted in stable conjugates and full antigenicity of the hapten antigen molecule. A commercial kit for cytokinin measurements was developed and new antiauxin (indole-3-acetic acid (IAA)) monoclonal antibodies (Mab) were produced.
A specific Mab recognizing an arabinogalactan protein eptiope in the plasma membrane of higher plants was developed and characterized chemically. This antibody appears to be a specific marker of the sporophytic phase of plant development but is excluded from all germ tissue. By coimmunization with sugar beet protoplasts and carrot antiserum, a new family of cell surface specific antibodies was subsequently developed, some of which may be able to recognize discrete steps in plant cell differentiation.
THE PROJECT WILL FOCUS ON BASIC AND APPLIED RESEARCH DIRECTED TOWARDS THE PLANT CELL SURFACE IN A MODEL SYSTEM (CARROT) AND A SOCIO-ECONOMIC CROP (SUGAR BEET). THE PRIMARY AIM WILL BE TO ESTABLISH ANALYTICAL METHODS TO BE USED IN THE CONTROL OF DIFFERENTIATION AND REGENERATION OF CELLS INTO WHOLE PLANTS, AS WELL AS IN CELL-CELL INTERACTIONS. THE WORK BE DISTRIBUTED BETWEEN LABORATORY A (DDS), LABORATORY B (NORWICH) AND LABORATORY C (COPENHAGEN).
IN LABORATORY A BASIC AS WELL AS EXPERIMENTAL IN VITRO SYSTEMS WILL BE FURTHER ELABORATED WITH MAIN EMPHASIS ON CELL AND TISSUE LEVEL. FURTHER TO THAT, EMPHASIS WILL BE LAID ON IN VITRO ASPECTS OF HOST-PATHOGEN INTERACTIONS (A).GRADUALLY DATA FROM BIOCHEMICAL AND IMMUNOLOGICAL STUDIES IN (B) AND (C) AT THE SUBCELLULAR LEVEL WILL BE INTEGRATED. THE PROPOSED EXTENSIVE ANALYSIS OF CELL SURFACE COMPONENTS (B) WILL DRAW HEAVILY ON TRANSFER OF TECHNOLOGY FROM (A) EMERGING DATA FROM (B) WILL BE INTEGRATED IN THE ATTEMPTS BY (C) TO EXTEND THEIR RESULTS INTO APPLICABLE IMMUNOCYTOCHEMICAL METHODS. IT IS ENVISAGED THAT RESULTS FROM THE CELL-CELL INTERACTION RESEARCH PROPOSED BY (B) WILL BE OF GREAT IMPORTANCE TO A DETAILED RESEARCH/DEVELOPMENT PROGRAMME ON DISEASE AND HERBICIDE RESISTANCE (A).
Funding SchemeCSC - Cost-sharing contracts