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 –“Good” for growth and rust, but variation among parents 70 full-sib families –Partial diallel with approx 4 to 5 crosses per parent –Intent is to go to the field with 60 FS fams

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. Shoot Collection

16. Preparing Cuttings To Set

18. 30 Clones Per Tray

19. Typical Rooted Cutting ( 9 Weeks from setting)

20. 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

21. Summary of Rooting Trial# of families# of clones # plots per clone # cuttings per plot Mean rooting % Range of fam. means May 7021944454%27-76% July 7021855438%18-69%

22. Variation Within Family for Rooting

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

24. 64% rooting81% rooting48% rooting Differences in Shoot Morphology Do these differences have an effect on rooting?

25. Differences in Root Morphology

26. ‘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

27. Disease Screening 1400 clones from May and July setting sent to RSC –22,000 rooted cuttings –5 to 20 ramets per clones –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 broad inoculum –Group 4: PC with narrow inoculum Measure phenotypes (disease symptoms): –1400 clones –Two very different pathosytems

28. Resistance Screening at USFS - RSC

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. Field Locations Six Locations in FL and GA Design at each location: –2 silvicultural treatments (HI and LO) –4 complete blocks per treatment –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 Two settings: Jan and April –Jan setting used for 3 sites –April setting used for three sites Field planting in Dec 2002

31. 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????

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