CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites)

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

Breeding

• 30 top loblolly parents – Half from coastal plain; half from Florida

• 70 full-sib families (go to field with 60 FS fams)

1 2 3 4 5 6 … 29 30

1 X X

2 X X

3 X X

4 X X

5 X

6

…

29 X

30 X X

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

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

Hedged FBRC Study B seedlings 6 weeks after hedging.

Close-up of individual hedgesix weeks after hedging.

Close-up of individual hedge twelve weeks after hedging.

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

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

Clonal hedges were randomized in April 2001.


• 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

May 7, 2001-61 weeks after sowing

-46 weeks after initial topping of seedling

-11 weeks after last hedging

July

Hedge Production

0

20

40

60

80

100

120

May-01

Jul-01

Jan-02

May-02

Setting

Ave

rage

Sho

ots

per

Hed

ge

May 2002

Shoot Collection

Preparing Cuttings To Set

30 Clones Per Tray

Attack of the Clones:Importance of Labeling and Organization

Typical Rooted Cutting(9 Weeks from setting)

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

Rooting

0

10

20

30

40

50

60

May-01 Jul-01 Jan-02 May-02

Setting

Roo

ting

%

• Averaging about 50%

• Heritable

• Tremendous within family variation

Variation Within Family for Rooting

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

100%

Clone #

July

Roo

ting

%

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

64% rooting 81% rooting48% rooting

Differences in Shoot Morphology

Clonal Differences

Differences in Root Morphology







Locations of 2002 Series 1 loblolly tests

Rust Hazard

Cooperator Low High

IPSanta Rosa Co.,

FL

RAYWest Nassau

Co., FL

WEY Americus, GA

PC Palatka, FL

MEAD Cuthbert, GA

UGA (with help)B.F. Grant Forest, GA

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

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????

Acknowledgements

Forest Biology Research Cooperative

Special thanks to International Paper

Physiology of Clones

Pre-Dawn Water Potential Measurements

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

Resistance Screening at USFS - RSC

Data Structure

Family Clone Phenotypic Data from HIGH Phenotypic Data from LOW SNP

1 1 y111 y211 y311 y411 y511 y611 y711 y811 S1S1

1 2 S1S2

1 14 S2S2

1 15 S1S2

2 1

2 2

2 14

2 15

59 1

59 2

59 14

59 15

60 1

60 2

60 14

60 15

Documents

CCLONES - ADEPT ( Comparing Clonal Lines On Experimental Sites)