CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites)

1. CCLONES - ADEPT(Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

2. ‘Series 1’ CCLONES Schedule • Breeding 1996-1997 • Sow seed March 2000 • Top seedlings June 2000 • Transplant seedling hedges Sept 2000 • Randomize hedges on pad April 2001 • Stick cuttings for rooting assessment1 May 2001 • Stick cuttings for rooting assessment2 July 2001 • Stick cuttings for Study B field tests Jan & May 2002 • Screen 1400 clones for rust and PC 2002 • Plant 915 clones on six locations December 2002 • Measure phenotypes 2003

3. Breeding • 30 top loblolly parents • Half from coastal plain; half from Florida • 70 full-sib families (go to field with 60 FS fams)

4. 32 elite parents crossed in partial diallel to create ~2200 clones from 70 full-sib families Seed stratified: 1/24/00 Seed sown: 3/3/00

5. Study B seedlings after hedging (left) and prior to hedging (right) Seedlings were hedged in June 2000.

6. Hedged FBRC Study B seedlings 6 weeks after hedging.

7. Close-up of individual hedge six weeks after hedging.

8. Close-up of individual hedge twelve weeks after hedging.

9. Hedges moved to 20,000 sq ft hedge-pad after transplanting

10. Experimental Design • Randomization • Clonal hedges were completely randomized on the hedge pad prior to setting • Fixed-tray system (135 cells) • Trays could then be randomized within each rep

11. Clonal hedges were randomized in April 2001.

12. ‘Series 1’ CCLONES Schedule • Sow seed March 2000 • Top seedlings June 2000 • Transplant seedling hedges Sept 2000 • Randomize hedges on pad April 2001 • Stick cuttings for rooting assessment1 May 2001 • Stick cuttings for rooting assessment2 July 2001 • Stick cuttings for Study B field tests Jan & May 2002 • Screen 1400 clones for rust and PC 2002 • Plant 915 clones on six locations December 2002 • Measure phenotypes 2003

13. May 7, 2001 -61 weeks after sowing -46 weeks after initial topping of seedling -11 weeks after last hedging

14. July

15. Hedge Production May 2002

16. Shoot Collection

17. Preparing Cuttings To Set

19. 30 Clones Per Tray

20. Attack of the Clones:Importance of Labeling and Organization

21. Typical Rooted Cutting(9 Weeks from setting)

22. Root AssessmentExperimental Design • May 2001 setting • Set ~2200 clones, 4 replications with 4-ramet row plots • Assessed rooting 9 weeks after setting • Counted # newly emerging roots from plug 9 weeks after setting • Shoot dry weights obtained from 1 ramet per clone per rep (3 reps) • Variance components estimated with ASREML • July 2001 setting • Set ~2200 clones, 5 replications with 4-ramet row plots • Assessed rooting 9 weeks after setting • Measured cutting diameter and height 9 weeks after setting (3 reps) • Variance components estimated with ASREML • January 2002 setting • May 2002 setting

23. Rooting • Averaging about 50% • Heritable • Tremendous within family variation

24. Variation Within Family for Rooting

25. Heritability Estimates For Root Number and Rooting % D/A = 0.16 D/A = 0.05 D/A = 0.14

26. Differences in Shoot Morphology 64% rooting 48% rooting 81% rooting

27. Clonal Differences

28. Differences in Root Morphology

29. ‘Series 1’ CCLONES Schedule • Breeding 1996-1997 • Sow seed March 2000 • Top seedlings June 2000 • Transplant seedling hedges Sept 2000 • Randomize hedges on pad April 2001 • Stick cuttings for rooting assessment1 May 2001 • Stick cuttings for rooting assessment2 July 2001 • Stick cuttings for Study B field tests Jan & May 2002 • Screen 1400 clones for rust and PC 2002 • Plant 915 clones on six locations December 2002 • Measure phenotypes 2003

30. ‘Series 1’ CCLONES Schedule • Breeding 1996-1997 • Sow seed March 2000 • Top seedlings June 2000 • Transplant seedling hedges Sept 2000 • Randomize hedges on pad April 2001 • Stick cuttings for rooting assessment1 May 2001 • Stick cuttings for rooting assessment2 July 2001 • Stick cuttings for Study B field tests Jan & May 2002 • Screen 1400 clones for rust and PC 2002 • Plant 915 clones on six locations December 2002 • Measure phenotypes 2003

31. Locations of 2002 Series 1 loblolly tests

32. Field Locations • Six Locations in FL and GA • Design at each location: • 2 silvicultural treatments (HI and LO) • 4 complete blocks per treatment; alpha-lattice • Total of 8 ramets per clone (2 x 4) per site • 915 clones from 60 FS families • Total size approx 14 acres • Total trees: 6 sites x 2 trts x 4 blocks x 915 clones = 44,000 • Three settings: Jan, May and July • Jan setting used for 0 - 1 site • May setting used for 3 sites • July setting used for 1-2 sites • Field planting in Dec 2002

33. Phenotyping of Association Pop’n • Rooting • May and July 2001 • Jan and April 2002 • Disease symptoms • Rust and PC in RSC • Rust in HI and LO treatments in field • Standard growth:1, 2, 3 height in HI and LO • Water deficit symptoms: 2 of 6 sites; 600 clones • Stable carbon isotopes at end of 1st season • Specific leaf area • Relative water content – two dry periods • Water potential – two dry periods • Other????

34. Acknowledgements Forest Biology Research Cooperative Special thanks to International Paper

35. Physiology of Clones

36. Pre-Dawn Water Potential Measurements

37. Disease Screening • 1400 clones from May and July setting sent to RSC • 22,000 rooted cuttings • 5 to 20 ramets per clone • Good rooting clones with approx equal numbers per family • Four groups (with 5 or less ramets per clones) • Group 1: Rust with broad inoculum • Group 2: Rust with narrow inoculum • Group 3: PC with RSC protocols • Group 4: PC with UF protocols • Each group in 5 blocks with 1 ramet per clone; varying #’s of clones • Measure phenotypes (disease symptoms): • 1400 clones • Two very different pathosystems

38. Resistance Screening at USFS - RSC

39. Data Structure