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

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

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

CCLONES - ADEPT(Comparing Clonal Lines On Experimental Sites)

Forest Biology Research Cooperative

University of Florida

Page 2: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

‘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

Page 3: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 4: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 5: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 6: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Hedged FBRC Study B seedlings 6 weeks after hedging.

Page 7: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Close-up of individual hedgesix weeks after hedging.

Page 8: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Close-up of individual hedge twelve weeks after hedging.

Page 9: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 10: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 11: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Clonal hedges were randomized in April 2001.

Page 12: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

‘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

Page 13: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

May 7, 2001-61 weeks after sowing

-46 weeks after initial topping of seedling

-11 weeks after last hedging

Page 14: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

July

Page 15: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Shoot Collection

Page 16: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Preparing Cuttings To Set

Page 17: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida
Page 18: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

30 Clones Per Tray

Page 19: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Typical Rooted Cutting(9 Weeks from setting)

Page 20: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 21: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Summary of Rooting

Trial # of families # of clones# plots per

clone# cuttings per plot

Mean rooting %

Range of fam.

means

May 70 2194 4 4 54% 27-76%

July 70 2185 5 4 38% 18-69%

Page 22: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Variation Within Family for Rooting

0%10%20%30%40%50%60%70%80%90%

100%

Clone #

July

Roo

ting

%

Page 23: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Heritability Estimates For Root Number and Rooting %

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

Her

itab

ility

Root # May RT % July RT %

Trait Measured

DominanceAdditive

D/A = 0.16

D/A = 0.05

D/A = 0.14

Page 24: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

64% rooting 81% rooting48% rooting

Differences in Shoot Morphology

Do these differences have an effect on rooting?

Page 25: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Differences in Root Morphology

Page 26: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

‘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

Page 27: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 28: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Resistance Screening at USFS - RSC

Page 29: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

‘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

Page 30: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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

Page 31: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

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????

Page 32: CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites) Forest Biology Research Cooperative University of Florida

Acknowledgements

Forest Biology Research Cooperative

Special thanks to International Paper