2. SABBI Barley Program ●SMART breeding (Selection with Markers and Advanced Reproductive Technologies) ●Conventional breeding ●Doubled haploid production ●Marker assisted selection

3. Program Goal Develop SAB brand aligned barley varieties with superior: ●Yield potential ●Stability ●Agronomic characteristics ●Disease resistance ●Malting quality ●Brewing quality for the Southern Cape dry land area and the Northern Cape irrigation areas

4. Southern Cape: Dry land ± 150 000 tons Northern Cape: Irrigation ± 70 000 tons Barley producing areas of South Africa

5. Research facility near Caledon in Southern Cape

6. Trial and F-generation material under irrigation

7. Phase 1: Crossing Collect germplasm Making crosses Phase 2: Selection in F-generations Phase 3: Trial evaluation Phase 4: Commercial evaluation Phase 5: Seed production and maintenance Conventional breeding Program

8. Crosses F1 F2 F3 F4 F5 Elite 1 Elite 2 Elite 3 LE 1 LE 2 LE 3 LE 4 LE 5 Comm Yr 1 Comm Yr 2 Comm Yr 3 Comm Yr 4 Seed multiplication Commercial production 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Limited quality evaluation Micro malting Experimental release Final release 4000 lines SABBI Barley Program

9. Collect germ plasm Germ plasm are collected from all over the world to address the deficiencies in the program Conventional breeding Program Phase1: Crossing

10. Most important characteristics Agronomic characteristics Brewing quality Malt quality Yield Plumpness Kernel nitrogen Maturity Straw length Straw strength Disease resistance Fermentability Brewing yield Starch gelatinisation temp Filterability Haze potential Taste stability Extract Kolbach Index FAN Diastatic activity Viscosity AAL Beta glucan

11. Making crosses Conventional breeding Program Phase1: Crossing

12. Plant breeding parents

13. Cross breeding parents

14. Phase 2: Selection in F-generations Conventional breeding Program

15. Phase 2: Selection in F-generations

16. Crosses F1 F2 F3 F4 F5 Elite 1 Elite 2 Elite 3 LE 1 LE 2 LE 3 LE 4 LE 5 Comm Yr 1 Comm Yr 2 Comm Yr 3 Comm Yr 4 Seed multiplication Commercial production 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Limited quality evaluation Micro malting Experimental release Final release 4000 lines 5000 lines 4000 lines 3000 lines SABBI Barley Program

17. Single plant selection in wide rows from F2 to F4 generation for characteristics like tillering, straw strength and height, maturity and disease resistance; single row selection in F5 on same characteristics to be proceeded to Evaluation phase

18. Conventional breeding Program Phase 3: Trial evaluation

19. Crosses F1 F2 F3 F4 F5 Elite 1 Elite 2 Elite 3 LE 1 LE 2 LE 3 LE 4 LE 5 Comm Yr 1 Comm Yr 2 Comm Yr 3 Comm Yr 4 Seed multiplication Commercial production 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Experimental release Final release 1000 lines 1 location 180 lines2 locations 46 lines2 locations 25 lines12 locations Micro malting SABBI Barley Program

20. Plant blocks at different localities and evaluate for yield, plumpness, kernel nitrogen, germination energy, disease resistance and malting quality (Extract, KI, Viscosity, AAL, beta-glucans, FAN, DP)

21. Conventional breeding Program Phase 4: Commercial evaluation

22. Crosses F1 F2 F3 F4 F5 Elite 1 Elite 2 Elite 3 LE 1 LE 2 LE 3 LE 4 LE 5 Comm Yr 1 Comm Yr 2 Comm Yr 3 Comm Yr 4 Seed multiplication Commercial production 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Experimental release Final release SABBI Barley Program

23. After experimental release varieties are planted by commercial producers for SABM and SAB to evaluate for malting and brewing quality for three years: Year 1, 350t Year 2, 1000t Year 3, 3000t

24. Conventional breeding Program Phase 5: Seed production

25. Crosses F1 F2 F3 F4 F5 Elite 1 Elite 2 Elite 3 LE 1 LE 2 LE 3 LE 4 LE 5 Comm Yr 1 Comm Yr 2 Comm Yr 3 Comm Yr 4 Seed multiplication Commercial production 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Experimental release Final release SABBI Barley Program

26. In the Southern Cape Breeder and Foundation seed are produced. Foundation seed are sold to Agricultural businesses who produce Certified seed to be sold to the producers. Under irrigation we are involved in the whole seed chain.

27. Doubled Haploid Production Marker Assisted Selection Extension in program SABBI Barley Program

28. Doubled Haploid production Marker Assisted Selection SABBI Barley Program Extension in program

29. Crosses F1 F2 F3 F4 F5 Elite 1 Elite 2 Elite 3 LE 1 LE 2 LE 3 LE 4 LE 5 Comm Yr 1 Comm Yr 2 Comm Yr 3 Comm Yr 4 Seed multiplication Commercial production 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Limited quality evaluation Micro malting Experimental release Final release Doubled Haploids Marker assisted selection SABBI Barley Program

30. Doubled haploid production ●The production of true breeding lines within one year to shorten the release time of new cultivars with three to four years ●F1 seed (and sometimes F2 seed) from selected crosses are used to produce Doubled haploid plants

31. Double haploid production Microspore method

32. Double haploid production Anthers plated on mediumCalli develop from anthersCalli transplanted onto regeneration medium and doubled haploid plants develop Anther culture method

33. Marker assisted selection ●The early identification of specific characteristics in the progeny of a specific cross. ●Especially important for characteristics which is difficult to identify or measure in a conventional way ●Indirect selection process through the ID of a unique DNA sequence occurring in proximity to a desired gene, by molecular techniques ●Contracted to CENGEN

34. Marker Assisted Selection TraitChromosomeOrigin Grain brightness3HFranklin Grain brightness2HHarrington Malt extract, diastatic power, α- amylase, free α-amino nitrogen, β-glucan 1HAlexis α-amylase5HLHarrington Diastatic power4HLHarrington Diastatic power5HLHarrington Wort content (sucrose)4HLDerkado Example of markers developed for specific traits