SEED ORCHARD CONFERENCE - uni-freiburg.deSeed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden Spatial design Clonal Seed Orchard of Wild Cherry – Constitution and Design

1

Research Institute for Nature and ForestResearch Institute for Nature and ForestGeraardsbergen, Belgium

www.inbo.be

SSEEDEED OORCHARDRCHARD CCONFERENCEONFERENCE

A New Generation of A New Generation of ClonalClonal Seed Orchards of Seed Orchards of

Wild CherryWild Cherry

Bart De Cuyper

Selection of Clones and Selection of Clones and Spatial DesignSpatial Design

Research Institute for Nature and ForestResearch Institute for Nature and ForestGeraardsbergen, Belgium

www.inbo.be

SSEEDEED OORCHARDRCHARD CCONFERENCEONFERENCE

A New Generation of A New Generation of ClonalClonal Seed Orchards of Seed Orchards of

Wild CherryWild Cherry

Bart De Cuyper

About picky trees and stubborn bumble bees

2

Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden

WhyWhy wild wild cherrycherry ??

WhyWhy selectionselection and and breedingbreeding ??

WhyWhy seedseed orchardsorchards ??

and and ……. . howhow ??

ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design



High-grade timberPrices standing timber

DBH ~ 50 cm : 200 € / m³DBH ~ 80 cm : 500 € / m³

Potential alternative for poplar for afforestation of set-aside or abandoned farmland

Silvicultural importanceValuable component of mixed hardwood stands


and… “doing a bit of everything is the best excuse fordoing something pretty close to nothing”

3



Annual demand for FRM ~ 30 kg of seeds

Before the Council Directive 1999/105/EC• Certification of FRM non-compulsory• Demand for FRM amply met by supply• Quality of FRM often unknown and mostly very bad

October 23th, 2003: implementation of Council Directive in Flemish forest law• Certification of FRM becomes compulsory• Genetic quality of FRM has to be warranted• Additional subvention for (re)forestation is confined to

“recommended” (= indigenous) basic material




Demand

Indigenous basic material• FRM of category “source identified” not allowed for

forestry purposes• Two seed stands• One seed orchard

Supply

Seed stands (1/2)• “Vrebos” (0.28 ha) & “Rattenberg” (0.37 ha)• Low yield ~ 10 kg in good crop years• Harvest is not cost-effective: cost price = 4 x selling price


4



Seed stands (2/2)• “Vrebos” is likely to be deleted from the National Register• Low genetic diversity (16 SSR’s)

- root suckering: 402 individuals 71 genotypes




0 50 100 150 200x

-100

-50

0

50

y

0 50 100 150 200x

-100

-50

0

50

y

5



Seed stands (2/2)• “Vrebos” is likely to be deleted from the National Register• Low genetic diversity (13 SSR’s)

- root suckering: 402 individuals 64 genotypes- genetic distance < 0.23


• Cross-compatibility- Gametophytically: 20 different S-genotypes- Phenologically: flowering period of clusters to be assessed

• Indication of introgression of sweet cherry varieties(to be confirmed)



Seed orchard• “Mommedeel” (° 1988 - 0.82 ha)


• 65 genotypes selected in two populations• Moderate yield ~ 20 kg in good crop years• Recent severe dieback

- unsuitable site- rejection of grafts

6










No perspectives for selection of additional seed stands

Seed orchards• enhanced yield and ditto genetic quality of offspring (IF well

conceived)• possibility for intensive management• harvest less labour-intensive (i.e. low-cost)

7




Selection of constituents

Spatial design

Spatial isolation

Management




Selection of constituents

SpatialSpatialSpatial designdesigndesign

SpatialSpatialSpatial isolationisolationisolation

ManagementManagementManagement

8


SelectionSelection of of clonesclones


Source = basic collection of 157 plus trees selected in 27 populations in the early 80’s

Vegetative replica of plus trees planted in 6 multiclonalplantations

Seed harvest in multiclonal plantations

Establishment of 13 half-sib progeny tests (Σ = 15.3 ha)

Assessment of h²A & General Combining Ability

Construction of selection indexn

Is = Σ ( h²A * GCA )ii = 1

with n = number of traits assessed




SourceSourceSource === basicbasicbasic collectioncollectioncollection of 157 plus trees of 157 plus trees of 157 plus trees selectedselectedselected in in in 27 27 27 populationspopulationspopulations

VegetativeVegetativeVegetative replica of plus trees replica of plus trees replica of plus trees plantedplantedplanted in 6 in 6 in 6 multiclonalmulticlonalmulticlonalplantationsplantationsplantations

SeedSeedSeed harvestharvestharvest in in in multiclonalmulticlonalmulticlonal plantationsplantationsplantations

Establishment of 13 Establishment of 13 Establishment of 13 halfhalfhalf---sibsibsib progenyprogenyprogeny tests (tests (tests (ΣΣΣ = 15.3 ha)= 15.3 ha)= 15.3 ha)

AssessmentAssessmentAssessment of of of hhh²²²AAA & General & General & General CombiningCombiningCombining AbilityAbilityAbility

Construction of Construction of Construction of selectionselectionselection indexindexindexnnn

IIIsss = = = ΣΣΣ ( ( ( hhh²²²AAA * GCA )* GCA )* GCA )iiii = 1i = 1i = 1

withwithwith n = n = n = numbernumbernumber ofofof traitstraits assessedassessed

9




Selection criteria• Vigour (height growth)• Morphology

- stem straightness- branching habit (number, tickness, angle of branches)- apical dominance

• Phenology- flushing- bud set- St. Johns sprouts

• Disease resistance




Selection criteria• Disease resistance

- anthracnosis Blumeriella jaapii (observation of natural infections)

- bacterial canker PseudomonasPseudomonas syringaesyringae pvpv. . syringaesyringae / / morsprunorummorsprunorum / / aviiavii (artificial infections)

10




SelectionSelectionSelection of of of constituentsconstituentsconstituents

Spatial design




SpatialSpatial designdesign


Wild cherry is entomophilous

Patch-like pollination pattern is suspected• Test case: multiclonal plantation (105 genotypes)• Parenthood analysis

- selection of 16 equally distributed mother trees- ad random selection of 60 seedlings in the half-sib offspring of each

mother tree- identification of fathers using

16 SSR’s (DP 0.76 0.95)SI-genotype (DP=0.98)

Pollen vectors = mainly bumble bees• Leave a scent mark on flowers they visit• Tend to forage on small groups of trees

11




Parenthood analysis

0

5

10

15

20

25

30

35

40

45

Shar

e of

pol

len

(%)

5 10 15 20 25 30 35 40

Distance from mother tree (m)

• Internal pollen flux: 75% of the pollen donors within a distance of 10 m from the mother tree (= 2 x planting distance)

• Paternal contribution was influenced by flowering period and S-(in)compatibility but not by flowering abundancy

• Exclusion method• SSR’s: Parent exlusion

probability > 0.99

• 21 % of the fathers are located outside the plantation




Neighbouring trees in future seed orchards should becross-compatible• phenologically: overlap of flowering period• gametophytically: different SI-genotype

12




Phenological compatibility• Observation of flowering period

- per stratum : observation of share of fully opened flowers every 48 hours for more then one month

- crown divided into 4 strata

• Observation in 4 plantations

• Observation in 2002 and 2004




Phenological compatibility• Observation of flowering period

0

10

20

30

40

50

60

70

80

90

100

0 48 120 168 216 312 384 504 720 888

Time since first observation (hours)

Shar

e of

fully

ope

ned

flow

ers

(%)

13




Phenological compatibility• however…

- fully opened flowers only receptive for 38 hours- viability of pollen in pollen bags of bumble bees = 12 hours

0

5

10

15

20

25

30

35

0 48 96 144 192 240 288 336 384 432 480 528 576 624 672 720 768 816 864 912

T ime s ince f irst o bservat io n (ho urs)




Phenological compatibility• however…

- fully opened flowers only receptive for 38 hours- viability of pollen in pollen bags of bumble bees = 12 hours

02468

101214161820

0 48 96 144

192

240

288

336

384

432

480

501

528

576

624

672

720

768

816

864

912

Time of observation (hrs)

Rec

eptiv

e flo

wer

s (%

)

Accession 48Accession 69Overlap

8 % 02468

10121416

0 48 96 144

192

240

288

336

384

432

480

528

576

624

672

720

768

Time of observation (hrs)

Rec

eptiv

e flo

wer

s (%

) Accession 76Accession 5Overlap

80 %

14




Phenological compatibility• furthermore…

- no influence of exposure or location within the plantation

N




Phenological compatibility• furthermore…

- no influence of exposure or location within the plantation- no significant intra-clonal variation- no significant differences between sites- observations of 2002 & 2004 are concordant- and…btw…~ 51,000 observations

15




Gametophytic compatibility• Governed by a multiallelic & highly polymorhic S-locus

(13 S-alleles in sweet cherry cv’s + 11 S-alleles in wild cherry)• Individuals with the same S-genotype are cross-incompatible• Molecular identification of S-alleles




Gametophytic compatibility – Molecular assessment• sizing of introns

16



Intron 1 Intron 2

PaSPcons-F

PaC1cons-R

HEX

PaC5cons-R

C3 C4 C5SP C1 C2 HV

PaC2cons-F

145 368 bp 580 2380 bp




Gametophytic compatibility – Molecular assessment• sizing of introns• allele-specific PCR

17




Gametophytic compatibility – Confirmation• controlled crosses




Major spin-off• parenthood analysis ~ incomplete diallel cross• determination of SCA for 51 combinations of clones• neighbouring trees should not only be cross-compatible but

also have high SCA-values

18






Spatial isolation



SpatialSpatial isolationisolation


Minimizing genetic pollution

21 % of the pollen is coming from outside the plantation

4%17%79%

400 m

19







Management


Establishment & ManagementEstablishment & Management


Vegetative propagation of mother trees• grafting on dwarfing rootstock• “Gisela 5” : compromis between dwarfing / compatibility

Planting in espalier

Bird netsBumble bee nestsInduce early and abundant flowering• pruning• tearing and bending of branches• use of herbicides• cutting of roots

20


StaffStaff


Breeding• Bart De Cuyper – Senior researcher• An Van Breusegem – Lab assistant• Wim Stevens – Field technician• Steven Haelterman – Field technician

Pathology• Marijke Steenackers – Senior researcher• Sabrina Neyrinck – Lab assistant• Kurt Schamp - Field technician

Documents

SEED ORCHARD CONFERENCE - uni-freiburg.deSeed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden Spatial design Clonal Seed Orchard of Wild Cherry – Constitution and Design