Mycotoxin Prevention in Cereal Crops by Enhanced Host Plant Resistance an update October2008
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1 Mycotoxin Prevention in Cereal Crops by Enhanced Host Plant Resistance an update October2008 UMB; Åsmund Bjørnstad, Helge Skinnes, et al BIOFORSK: Sonja Klemsdal, Erik Lysøe et al. UMN: Jim Anderson, Howard Rines, Ruth Dill-Macky, Brian Steffenson
2 Norwegian team/projects on Fusarium: UMB: Helge Skinnes, Åsmund Bjørnstad, post docs, PhDs, MSc NFR project Safe grains ( , Marker-assisted selection ( , EEA Hungary, Ethiopia Nordic proposal on oat genomics BIOFORSK: Sonja Klemsdal, Guro Brodal, post docs, PhDs NFR projects: Reduced risk of Fusarium and mycotoxin contamination Mycotoxin contamination in oats Genes involved in the infection process and biosynthesis of mycotoxins
3 NORWEGIAN UNIVERSITY OF LIFE SCIENCES University of Minnesota Wheat & Barley Scab Researchers Department of Agronomy & Plant Genetics Jim Anderson, Wheat Breeding Kevin Smith, Barley Breeding Gary Muehlbauer, Wheat & Barley Mol. Genetics Dave Garvin, USDA-ARS Wheat Genetics Department of Plant Pathology Ruth Dill-Macky, Small grains Brian Steffenson, cereals & wild relatives Yanhong Dong, Mycotoxin Analyses Corby Kistler, USDA-ARS Yue Jin, USDA-ARS Char Hollingsworth, Crookston Funding: National Science Foundation USDA-NRI Plant Genome Program U.S. Wheat and Barley Scab Initiative, via USDAARS Minnesota Small Grains Initiative (Minnesota Agricultural Experiment Station) U.S. Wheat & Barley Scab Initiative
4 NORWEGIAN UNIVERSITY OF LIFE SCIENCES Fusarium Head Blight (Scab) Evaluat Disease Severity Grain Soundness
5 NORWEGIAN UNIVERSITY OF LIFE SCIENCES DNA Marker Screening Tissue Harvest Grinding PCR prep Gel Loading
6 NORWEGIAN UNIVERSITY OF LIFE SCIENCES Dissecting a Genetically Complex Trait Resistant Susceptible % FHB Severity Homozygous Recombinants
7 Marker Assisted Evaluation of FHB Resistance : good diagnostic markers for the gene Fhb1 Xgwm389 alleles Sumai 3 One diagnostic allele! No. f o Crop Sci. 43: (2003)
8 DNA Marker Mapping cm Association of DNA markers (and linked genes) with scab resistance 14.2 LOD 3.0 No Associatio n 0.0 bcd907r gwm389 bcd907z BARC75 SUN2 gwm533a gwm493 BARC87 BARC102 gwm533b fba91 BARC101C BARC73 gwm274d
9 Narrow Closing down the in QTL on region Cloning a Scab Resistance Gene Marker Xgwm533 Marker Xgwm cm Qfhs.ndsu-3BS Qfhs.ndsu-3BS 0.3 cm 0.08 cm 7 candidate genes Xgwm493 Xgwm493 Fall 2003 Fall 2004 Fall 2005 Fall 2008
10 NORWEGIAN UNIVERSITY OF LIFE SCIENCES UMN10: a perfect diagnostic marker now in use in Norwegian wheat breeding
11 NORWEGIAN UNIVERSITY OF LIFE SCIENCES UMB: Avoidance of Fusarium through anther extrusion Line 20055: susceptible Line 20054: resistant Parent Arina: moderate Parent NK: moderate
12 NORWEGIAN UNIVERSITY OF LIFE SCIENCES If anthers are thrown out, low DON levels! Regression of DON in 2002 on anther extrution for Arina x NK93604 y = 29,777e -0,2427x R 2 = 0,2823 DON Arina 10 0 NK AE
13 Fusarium in oats NORWEGIAN UNIVERSITY OF LIFE SCIENCES
14 NORWEGIAN UNIVERSITY OF LIFE SCIENCES Infected seeds in the cv. Hurdal inoculated with F. graminearum: Discoloration and shrivelling
15 NORWEGIAN UNIVERSITY OF LIFE SCIENCES Seed infection in oats: left susceptible cultivar, right resistant but do DON levels correspond?
16 Resistance appears highly heritable ( Hurdal x Z615-x cross) (In 2008 both in UMB and UMN) Freezer blotter test after spray inoculation in field 2006 at Aas by Fusarium cumorum mixture. 25 susceptible resistant 5 0 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 Hurdal/Z615-4 HURDAL BESSIN B605X % highly + medium infected seeds NORWEGIAN UNIVERSITY OF LIFE SCIENCES
17 NORWEGIAN UNIVERSITY OF LIFE SCIENCES Progress 2008, Hurdal x Z595-7 cross 184 RIL lines, Mycotoxin content this autumn and 2009 DArT data are available 32 linkage groups were generated, comprising of 330 markers A set of 205 SSR markers screened here, ca in USDA lab in Aberdeen, Idaho (courtesy of prof. H. Rines, UMN) Nordic proposal on genomics in oat submitted
18 Entry pathways of Fusarium into oat florets (MSc thesis UMB/UMN ) Floret mouth is principal entry pathway (Apical infection) Anther colonization has important role at initial stage of infection process MSc student Selamawit Tekle Gobena 3 months internship in UMN 2008, prof. Ruth Dill-Macky visiting in UMB/Bioforsk 2 months this fall
19 Effect of time of Fusarium inoculation Most susceptible to infection around flowering Can be spread out in time, toxins can build up later also, but less kernel damage
20 Project at Bioforsk: Mycotoxin contamination in oat characterization of the infection process by the major T-2 / HT-2 toxin producer Fusarium langsethiae Project leader: Sonja Sletner Klemsdal, Bioforsk Researcher: Hege Hvattum Divon, Bioforsk
21 Infection route? Life cycle? No symptoms on the host or just microscopic? Pathogen or endophyte? Different host preferences? Different climatic preferences? Molecular mechanisms? T-2/HT-2 toxins
22 Main Objective: Enhance our understanding of fundamental processes of F. langsethiae colonization of oat. Reduce the mycotoxin content in oat. Mikkelsen et al., 2003 Generate a fluorescent marker strain Other infection experiments (possible collaboration with Ruth DillMacky 1 Characterization of F. langsethiae growth on the plant 2 The role of T-2/HT-2 toxins in F. langsethiae establishment on the plant Hph Tri5 Trichothecenes; T-2/HT-2 EST library Expression in planta Characterization of fungal genes involved in the colonization process 3 Hph Candidate gene EFFECT ON GROWTH IN PLANTA?
23 Project at Bioforsk: Reduced risk of Fusarium and mycotoxin contamination in Norwegian cereals by the development of a rapid screening system S.S. Klemsdal, O. Elen, I.S. Hofgaard, H.U. Aamot and G. Brodal Bioforsk - Norwegian Institute for Agricultural and Environmental Research, Plant Health and Plant Protection Division Ås, Norway
24 To reduce the risk of mycotoxin contaminated grain entering the food and feed processing chain, a screening system is under development: Prediction model(s), based on information on agronomic/cultivation practice and climatic conditions, will be used to estimate the risk of Fusarium/mycotoxin development in cereal fields 2 Grain from high-risk fields will be analysed for Fusarium and/or mycotoxins by a rapid screening method, before entering the mill 3 Samples from lots close to the defined maximum toxin limits (based on analyses in step 2) will be forwarded to chemical mycotoxin analyses A system to identify and avoid cereal grain lots with mycotoxin levels above the maximum risk limits
25 Project at Bioforsk: The transcription factor FgStuA influences spore development, pathogenicity and secondary metabolism in Fusarium graminearum Post doc: Erik Lysøe, Bioforsk Collaboration with Dr. H. Corby Kistler, Cereal Disease Laboratory, UMN
26 Conidia Ascospores Toxins Pigmentation Pathogenicity Chitin FgStuA Global regulator? Hydrophobicity Cell cycle Nitrogen regulation Oxidative stress
27 Teachers: Partners in the Nordic Research Network New emerging mycotoxins and secondary metabolites in toxigenic fungi Dr. Corby Kistler, Minnesota Dr. Todd Ward, USDA, Illinois
28 NORWEGIAN UNIVERSITY OF LIFE SCIENCES The current project collaboration (1): Marker exchanges in wheat and oats Thousands of UMB samples analyzed for toxins in UMN lab (Dr. Yanhong Dong) Post doc Erik Lysøe to UMN , Corby Kistler s lab Student exchange to UMN: MSc Janne Kvame 2007, MSc student Selamawit Tekle Gobena 2008 Dr. Corby Kistler teaching in UMB 2008 Visiting professorship for prof. Ruth Dill-Macky by UMB grants from October to December 2008 Visiting professorship for prof. Åsmund Bjørnstad by Fulbright and UMB grants Of increasing importance!
29 NORWEGIAN UNIVERSITY OF LIFE SCIENCES The current project collaboration (2): Papers Joint papers planned: Infection process in oats, to be published 2009 Erik Lysøe/Corby Kistler paper submitted by December
30 NORWEGIAN UNIVERSITY OF LIFE SCIENCES Questions and challenges Still: UMN funding for the collaboration! Plant/animal biotechnology course in UMB in 2009? The genomics projects in plants in UMN, how can the links be forged to UMB? To develop more on the health aspects in oats as a food (UMN medical School already has links with University of Oslo)
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