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

2. ‘Series 1’ CCLONES Schedule Breeding1996-1997 Sow seedMarch 2000 Top seedlingsJune 2000 Transplant seedling hedgesSept 2000 Randomize hedges on padApril 2001 Stick cuttings for rooting assessment1 May 2001 Stick cuttings for rooting assessment2July 2001 Stick cuttings for Study B field testsJan & May 2002 Screen 1400 clones for rust and PC2002 Plant 915 clones on six locationsDecember 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) 123456…2930 1XX 2XX 3XX 4XX 5X 6 … 29X 30XX

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

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 seedMarch 2000 Top seedlingsJune 2000 Transplant seedling hedgesSept 2000 Randomize hedges on padApril 2001 Stick cuttings for rooting assessment1 May 2001 Stick cuttings for rooting assessment2July 2001 Stick cuttings for Study B field testsJan & May 2002 Screen 1400 clones for rust and PC2002 Plant 915 clones on six locationsDecember 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 Assessment Experimental 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. 64% rooting81% rooting48% rooting Differences in Shoot Morphology

27. Clonal Differences

28. Differences in Root Morphology

29. ‘Series 1’ CCLONES Schedule Breeding1996-1997 Sow seedMarch 2000 Top seedlingsJune 2000 Transplant seedling hedgesSept 2000 Randomize hedges on padApril 2001 Stick cuttings for rooting assessment1 May 2001 Stick cuttings for rooting assessment2July 2001 Stick cuttings for Study B field testsJan & May 2002 Screen 1400 clones for rust and PC2002 Plant 915 clones on six locationsDecember 2002 Measure phenotypes 2003

30. ‘Series 1’ CCLONES Schedule Breeding1996-1997 Sow seedMarch 2000 Top seedlingsJune 2000 Transplant seedling hedgesSept 2000 Randomize hedges on padApril 2001 Stick cuttings for rooting assessment1 May 2001 Stick cuttings for rooting assessment2July 2001 Stick cuttings for Study B field testsJan & May 2002 Screen 1400 clones for rust and PC2002 Plant 915 clones on six locationsDecember 2002 Measure phenotypes 2003

31. Locations of 2002 Series 1 loblolly tests Rust Hazard CooperatorLowHigh IP Santa Rosa Co., FL RAY West Nassau Co., FL WEYAmericus, GA PCPalatka, FL MEADCuthbert, GA UGA (with help) B.F. Grant Forest, GA

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 1 st 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 FamilyClonePhenotypic Data from HIGHPhenotypic Data from LOWSNP 11y111y211y311y411y511y611y711y811S1S1 12S1S2 114S2S2 115S1S2 21 22 214 215 591 2 14 5915 601 2 14 6015