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511: Functional analysis and expression of sclerotial mycoparasitism genes

Reference	BBS/E/H/00JW0230
Principal Investigator / Supervisor	Professor John Michael Whipps
Co-Investigators / Co-Supervisors
Institution	University of Warwick
Department	Warwick HRI
Funding type	Research
Value (£)	232,700
Status	Completed
Type	Institute Project
Start date	01/04/2005
End date	31/03/2006
Duration	12 months

Abstract

"We are identifying unique pathogenicity genes and analysing their function and expression as the mycoparasite, Coniothyrium minitans, attacks sclerotia of the plant pathogen, Sclerotinia sclerotiorum. A number of putative pathogenicity genes involved in mycoparasitism have been identified by insertional mutagenesis and we need to confirm their function. We will develop high throughput gene silencing technologies for C. minitans involving RNAi and homologous integration (knockout/disruption) approaches. RNAi should prove an effective mechanism of down regulating gene expression and will expedite functional gene analysis. In some cases, down- regulation is insufficient to yield phenotypic change and there remains a need for alternative disruption technologies. Agrobacterium has proved particularly efficient at targeting homologous integration in fungi and this system will be tested in C. minitans along with protoplast mediated approaches. Other pathogenicity genes involved in complete colonisation of sclerotia have been identified through suppressive subtraction hybridisation (SSH). A subtracted cDNA library of 672 clones encoding candidate mycoparasitism genes has been established and putative functional annotation through bioinformatics is being done. However, identification of genes involved in the recognition, spore germination and attachment of the mycoparasite to the host as well as in the early colonisation processes is needed to provide a complete picture of the whole process of sclerotial mycoparasitism. To address this, two approaches will be employed: i) SSH analysis of the early sclerotial colonisation stage will be carried out and a SSH- cDNA library of up to 1000 clones will be established and ii) a cDNA library of C. minitans spores germinating in response to sclerotial interaction will be constructed and cryo-preserved with a view to utilising around 3000 clones for expression analysis. This will lead to microarray expression profiling."

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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