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Pure Lining Breeder Seed for Molecular Variety ID in Oat AOWC, July 23, 2006, Fargo

This article discusses the use of molecular characterization for identifying oat varieties and its relevance in different aspects such as PBR, variety registration, pedigreed seed, and commercial grain. It also explores the benefits and limitations of molecular purification compared to botanical characterization.

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Pure Lining Breeder Seed for Molecular Variety ID in Oat AOWC, July 23, 2006, Fargo

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  1. Pure Lining Breeder Seed for Molecular Variety ID in OatAOWC, July 23, 2006, Fargo Brian Rossnagel, Peter Eckstein, Tom Zatorski & Graham Scoles Crop Development Centre, University of Saskatchewan Crop Development Centre - Plant Sciences Department

  2. Different ID Objectives • PBR • Variety registration • Pedigreed Seed • Commercial Grain CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  3. PBR • Simple question - “Are you growing my variety?” • If not - obvious question of “What is it?” is NOT relevant • Rarely needed • DUS issues - later CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  4. Variety Registration - Pedigreed Seed • Must have botanical phenotype (description) for Pedigreed Seed • Makes sense to have same for Variety Registration • DUS - not at issue - OK for 70 years • Molecular characterization could be useful, but has not been necessary to date & should remain optional • CSGA system critical in Canada CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  5. Commercial Grain • Different ball game • Need to know what it is • Mixtures - what & quantified • Botanical phenotype useless • IP systems effective • Molecular characterization helpful - not ready yet CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  6. CDC Breeder Seed Purification • Bulk Inc. & 200 Panicle selection - 2004 • 200 Breeder Hills – 2005 – discard phenotypic outliers – visual, disease, quality, molecular • Breeder Long Rows/Breeder Seed - 2006 • Each Row traces to single panicle CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  7. CDC Weaver Breeder Long Rows CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  8. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  9. Molecular Purification • No different than botanical - lab not field but OBJECTIVE! • Less expensive & risky • No environmental variability • DUS - yes, yes & yes CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  10. DUS • D - Canadian Hulless Barley Pilot project • U - Breeder’s molecular purification • superior to botanical • S - Non issue • extremely low probability of change in band pattern in so few generations from Breeder Seed CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  11. CDC Weaver - Oat • 200 Breeder lines • 20 botanical discards • 12 molecular discards • 6 in common • Two had 4 & three had 2 non-conforming bands & were most different by visual phenotype CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  12. CDC Weaver Long Rows 1 2 3 4 5 6 1 2 3 4 5 6 7 Long Row 51 - Discarded 8 9 CDC WeaverLong Rows Bulked DNA Samples 1 AC Assiniboia 2 CDC Boyer 3 CDC Pacer 4 CDC Orrin 5 CDC Dancer 6 CDC Weaver CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  13. * 1 * 2 * 3 * CDC Weaver Bulk DNA * 4 * 5 6 * Segregates 50/50 in population 7 7 Long Row 51 - Discarded * 8 9 CDC WeaverLong Rows CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  14. 150 172 154 174 20 Botanical Discards 12 Molecular Discards 6 in common, including 150, 154, 174 CDC WeaverLong Rows CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  15. Canadian Hulless Barley Pilot Project • 23 currently registered varieties • 4 breeding programs • 2 row & 6 row • AFLP • 3 primer combinations tried - each able to separate all 23 CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  16. 1 2 3 4 5 6 7 8 9 * * * 10 * 11 12 * 13 bands (minimum) required to distinguish 23 varieties * * * 13 * Additional polymorphic bands * * HullessbarleyE3M3 23 Varieties CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  17. 12 bands (minimum) required to distinguish 23 varieties 1 2 3 HullessbarleyE5M8 4 5 6 7 23 Varieties 8 9 10 11 12 CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  18. AC Bacon AC Hawkeye CDC Dawn HB803 Falcon CDC Alamo CDC Fibar CDC Silky CDC Gainer CDC McGwire Jaeger Merlin CDC Freedom Alberte Tercel CDC Speedy Phoenix Peregrine Tyto CDC Candle CDC Rattan HB805 Condor Hulless barley E5M8 CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  19. Canadian Hulless Barley Pilot Project • every primer set won’t work as well • multitudes of primer sets to try • microsats = alternate choice CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  20. Botanical Vs Molecular Characterization for PBR • Subjective vs Objective • Don’t hold to different standards • Don’t hold to higher standard • Not for all crops • Not for all breeders • OPTION NOT Addition CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  21. Mol. Vs Bot. Changes • Changes to Breeder Purification • Breeder’s call = botanical description and variety name • Keeper of the Key - CFIA • Option NOT Addition • Costs CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

  22. Do we need a centralized approach? • If a breeder feels that a particular technique is reproducible and can establish DUS of their cultivar satisfactorily and is willing to take that risk – why not – who else loses? • Let’s keep in mind how often this information may be needed. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT

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