Objective
Genes conferring pathogenicity and avirulence in the fungal pathogen Cladosporium fulvum, the causal agent of tomato leaf mould, will be cloned. The function and regulation of those genes will be studied. The final objective is to clone the resistance genes in the tomato plant complementary to the fungal avirulence genes. Both resistance genes and avirulence genes can be exploited to obtain broad spectrum resistance to plant pathogens.
The objective of the project is the unraveling of the molecular basis of the plant-fungus interaction tomato-Cladosporium fulvum. Molecular characterization of gene-for-gene systems is in progress. Of the many bacterial genes and one fungal avirulence gene cloned to date none showed homology with other characterized genes. It is likely that the primary function of avirulence genes varies considerably. In some cases an avirulence gene may be so important for the pathogen that loss of it would be detrimental. A property all avirulence genes have in common is the antigenicity of their direct or indirect products. Through these products the pathogens are recognized by the host which responds by inducing a hypersensitive response (HR). The primary function of resistance genes is still a matter of speculation, as none have yet been cloned. Without knowing the primary function of either avirulence and resistance genes their secondary function (the induction ofHR by their interacting products) could perhaps be exploited to engineer transgenic disease-resistant plant.
Results of research to date include:
The first fungal avirulence gene, avr9, of C fulvum that fits a gene for gene relationship has been cloned. Cloning of another avirulence gene avr4 is within reach;
gene disruption has been successfully performed in C fulvum;
Transformation of C fulvum with the vector pAN8-1 has resulted in a number of mutants with reduced pathogenicity;
Two telomere sequences of C fulvum with the sequence TTAGGG have been isolated;
Tomato plants transgenic for chimeric beta 1,3-glucanase and chitinase genes have been obtained;
Apoplastic fluids of C fulvum-infected tomato leaves contain enzymes that release oligosaccharides from plant and fungal cell walls;
The cloning of a polygalacturonase inhibitor protein (PGIP) from Phaseolus vulgaris is completed.
Adopting an integrated approach involving biochemical, genetic and molecular techniques We shall investigate the structure, function and regulation of genes and gene products controlling the pathogenicity and race-specificity of C.fulvum and the resistance of tomato (lycopersicon esculentum). Two approaches will be employed. Firstly, the proteins isolated from apoplastic fluid of infected tomato leaves will be characterized. Their genes (whether fungal or plant) will be cloned and their functions determined by transformation of tomato or C.fulvum as appropriate. The model for this approach is the successful purification of the A9 fungal avirulence peptide and the cloning of its encoding gene. The role of apoplastic plant proteins such as 1,3-B-glucanases, chitinases and proteinaceous inhibitors of endopolygalacturonases as Well as oligosaccharides released by some of these enzymes will be studied. Also the structure and function of these oligosaccharides in inducing and enhancing defence responses will be objects of study. The second approach is an unbiassed search for fungal gene products controlling pathogenicity and avirulence. This Will involve characterization of non-pathogenic mutants created by integrative transformation and the use of promoter-probe vectors based on the B- glucuronidase reporter gene.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences microbiology mycology
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Coordinator
6709 PD WAGENINGEN
Netherlands
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