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KEYNOTE LECTURE 2 SESSION 4: GENETICS OF HOSTS – PLANT RESISTANCE TO FUSARIUM, VARIETY DEVELOPMENT Metabolo-proteomics approach to identify candidate genes for wheat resistance to fusarium head blight A. C. Kushalappa McGill University, Ste.-Anne-de-Bellevue, QC, H9X3V9, Canada E-mail: ajjamada.kushalappa@mcgill.ca The resistance in wheat and barley to fusarium head blight, caused by Fusarium graminearum, is quantitatively inherited. Hundreds of quantitative trait loci (QTLs) for resistance have been identified but these contain several genes, including undesirable ones due to linkage drag. Near isogenic lines, with contrasting alleles at QTL-Fhb1, were pathogen or mock-inoculated, metabolites and proteins were analyzed using high resolution mass spectrometry. Mass spectral outputs were processed using MZmine for metabolites and MASCOT for proteins. The abundances were used to identify resistance-related metabolites and proteins, and mapped to metabolic pathways. Metabolites of the shunt phenylpropanoid pathway such as hydroxycinnamic acid amides, phenolic glucosides and flavonoids were significantly induced in the resistant NIL. Concurrently, the enzymes of phenylpropanoid biosynthesis such as cinnamyl alcohol dehydrogenase, caffeoyl-CoA O-methyltransferase, caffeic acid Omethyltransferase, flavonoid O-methyltransferase, agmatine coumaroyltransferase and peroxidase were also up-regulated. A protein coding gene (GENBANK No: CBH32656.1) near the Fhb1 locus was putatively annotated as hydroxycinnamoyl transferase that catalyzes the conjugation of hydroxycinnamic acid amides, whose high expression in resistant NIL was confirmed by quantitative RT-PCR using the sequence of wheat agmatine coumaroyltransferase. This demonstrates the potential of metabolo-proteomics approach to identify biotic stress resistance candidate genes. This gene can be used in plant improvement following further validation. Similar analyses of NILs with QTL-5A also significantly induced hydroxycinnamic acid amides, especially p-coumaroyl putrescene, caffeoylputrescine and feruloyl tyramine, and related metabolites. Keywords: OMICs, metabolomics, quantitative resistance, Fusarium head blight 69
SESSION 4: GENETICS OF HOSTS – PLANT RESISTANCE TO FUSARIUM, VARIETY DEVELOPMENT Semi-dwarf ‘uzu’ barley carries enhanced resistance to a range of pathogens including Fusarium culmorum F. Doohan, L. Gunupuru, S. Ali E-mail: fiona.doohan@ucd.ie Brassinosteroids (BRs) are hormones that influence plant growth, development and defence responses. The BR receptor protein Brassinosteroid Insensitive 1 (BRI1) has been characterised in several plant species. Semi-dwarf barley ‘uzu’ varieties have a mutation in the kinase domain of Bri1. In studies conducted using barley genotypes Akashinriki and Bowman and their ‘uzu’ derivatives, we show ‘uzu’ is more resistant to Fusarium head blight (FHB) disease. Resistance was characterised as the uzu plants having > 38% lower disease symptoms and > 25% higher yield as compared to parent lines Akashinriki and Bowman. Seedling experiments showed that the ‘uzu’ derivative of Akashinriki was also 87% more resistant to Fusarium seedling blight as compared to the parent line. Virusinduced gene silencing of Bri1 lead to enhanced susceptibility of detached leaves to Fusarium culmorum (>2 times more disease than plants treated with the empty VIGS vector). Thus the Fusarium resistance of ‘uzu’ is not a consequence of reduced BRI1 activity. Microarray analysis identified genes differentially regulated between the mutant and wild type during disease development. We are currently determining the effect of the uzu mutation on downstream BR signalling and the functionality of the kinase domain of the uzu Bri1. Furthermore, studies on net blotch disease and Barley Stripe Mosaic Virus show that ‘uzu’ confers broadspectrum resistance. We now begin a four-year project in order to determine the factors underpinning the ‘uzu’ associated resistance. We want to determine why ‘uzu’ has enhanced resistance to Fusarium and if any of the genes identified via microarray of ‘uzu’/Akashinriki contribute to the Fusarium resistance inherent in ‘uzu’? Preliminary results show that the gene silencing of a membrane receptor up-regulated in ‘uzu’ versus Akashinriki in response to Fusarium results in enhanced Fusarium disease in detached leaf assays. Our second objective is to determine if genes identified in other studies as contributing to Fusarium resistance play a role in brassinosteroid signaling. Acknowledgements: Dr. Sato, Barley Germplasm Centre, Kurashiki, Japan for providing the Uzu and Akashinriki cultivar lines. Dr.Paul nicholson, JIC, Norwich for providing Bowman and Bowman-Uzu cultivar lines. This work has emanated from research conducted with the financial support of the Irish Department of Agriculture, Food and the Marine and Science Foundation Ireland. Keywords: Brassinosteroid, Fusarium, resistance 70
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Bordeaux EFS12 12 th European Fusar
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TABLE OF CONTENTS Welcome from the
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4 SPONSORSHIP The 12 th European Fu
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8 13:00 - 14:00 Lunch 14:00 - 15:00
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10 Session 4: Genetics of Hosts - P
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12 Thursday, May 16 th Session 6: F
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Evidence for birth-and-death evolut
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Future challenges of Fusarium and m
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de Hoog G. S., 89, 114, 236 de Kler
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BOKU-University of Natural Resource
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touatisihem03@yahoo.fr He Xinyao In
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steve.oliver@bioc.cam.ac. uk Ortega
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Subramaniam Gopal Agriculture and A
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