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Structure/function analysis of LRR proteins and their ligands in plant pathogen interactions and engineered resistance

Ziel



This consortium will carry out structure/function analysis of proteins that play a role, or that may play a role, in plant-pathogen interactions. Proteins carrying leucine-rich repeats (LRRs) and their interaction with other proteins, will be of primary interest. Our main scientific objective is to advance our understanding of plant-pathogen interactions at the level of structural biology and protein/protein interaction. In collaboration with an industrial partner (MOGEN), our applied research objective is to exploit Cf-Avr genetic interactions to deliver durable resistance to potato late blight caused by Phytophthora infestans. Longer term, our investigations will open up the useful possibility of engineering synthetic resistance genes that recognize defined pathogen-derived ligands.
We will study three classes of LRR proteins and (where appropriate) their interaction with molecules derived from fungal plant pathogens. (i) Proteins encoded by the tomato resistance genes Cf-4 and Cf-9, which carry 26 or 28 LRRs respectively, and their interaction with the AVR peptides AVR4 and AVR9, produced by the fungal pathogen Cladosporium fulvum. The Cf-gene products enable specific recognition of the AVR products and upon recognition, activate the defence response. We will carry out biochemical and structural analysis of the Cf proteins, and their potential AVR ligands. We will investigate whether the genetic interaction between Cf and Avr genes involves direct physical interaction of their gene products. Analysis of various mutant AVR peptides will help unravel the molecular and physical basis of ligand/receptor interaction.
(ii) Plant polygalacturonase inhibiting proteins (PGIPs), which carry 10 LRRs and inhibit fungal polygalacturonases (PGs). PGs have important functions during fungal pathogenesis, such as tissue maceration and release of nutrients. Inhibition of PG by PGIP might attenuate fungal pathogenicity and lead to the accumulation of oligogalacturonides that can activate plant defences. PGIPs, and PGIP/PG interactions will be studied. Fusarium moniliforme PG will be used. The crystal structure of bean PGIP and F. moniliforme PG will be determined, and we will attempt to define how they physically interact. Data obtained will provide insight into protein/protein recognition. A chimeric Cf-9/PGIP gene, in which the domain of PGIP that might recognize PG replaces the domain of Cf-9 that probably recognizes AVR9, will be tested for whether it can activate plant defences upon recognition of PG.
(iii) A small secreted LRR protein (LRP) of tomato, carrying 4 LRRs, and without a currently known function, which is induced by viroid infection. Its small size should enable the crystal structure of this class of LRR to be rapidly defined. Experiments to assign its function will be conducted. This basic research will underpin an applied research project. A two component system involving Cf and Avr genes, in which the Avr gene is under the control of a pathogeninducible promoter, could deliver broad spectrum disease resistance. Quantitative control of recognition and specificity in the interaction will be crucial to effectively engineering this system. MOGEN NV of the Netherlands will participate in this project by testing various Avr gene alleles developed by PdW to try and reduce this concept to practice. Additionally, by using the Cf-9 gene provided by JJ, they will be able to attempt to use this system to deliver resistance to P. infestans in potato. The consortium will bring together expertise from Holland, Spain, Italy and the UK, in fungal molecular genetics, plant molecular genetics, plant biochemistry, crystallography and NMR to advance understanding of host-pathogen recognition at a molecular level, and to exploit this understanding to engineer broad spectrum disease resistance.

Wissenschaftliches Gebiet (EuroSciVoc)

CORDIS klassifiziert Projekte mit EuroSciVoc, einer mehrsprachigen Taxonomie der Wissenschaftsbereiche, durch einen halbautomatischen Prozess, der auf Verfahren der Verarbeitung natürlicher Sprache beruht. Siehe: Das European Science Vocabulary.

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Koordinator

AMICA SCIENCE EEIG
EU-Beitrag
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Adresse
Norwich Research Park John Innes Centre, Colney
NR4 7UH NORWICH
Vereinigtes Königreich

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Beteiligte (6)

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