1. Pure Lining Breeder Seed for Molecular Variety ID in Oat AOWC, 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. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT Different ID Objectives  PBR  Variety registration  Pedigreed Seed  Commercial Grain

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

4. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

5. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

6. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

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

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

10. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

11. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

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

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

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

15. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

16. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT Hulless barley E3M3 * 1 2 3 4 5 6 7 8 9 11 12 10 13 * * * * * * * * * 23 Varieties 13 bands (minimum) required to distinguish 23 varieties Additional polymorphic bands *

17. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT Hulless barley E5M8 1 2 3 4 5 6 7 8 9 10 11 12 23 Varieties 12 bands (minimum) required to distinguish 23 varieties

18. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT Hulless barley E5M8 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

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

20. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

21. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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

22. CROP DEVELOPMENT CENTRE - PLANT SCIENCES DEPARTMENT 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.