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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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy wild wild cherrycherry ??
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
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
and… “doing a bit of everything is the best excuse fordoing something pretty close to nothing”
3
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy selectionselection and and breedingbreeding ??
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
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy selectionselection and and breedingbreeding ??
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
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
4
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy selectionselection and and breedingbreeding ??
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
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
0 50 100 150 200x
-100
-50
0
50
y
0 50 100 150 200x
-100
-50
0
50
y
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy selectionselection and and breedingbreeding ??
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
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
• 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 Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy selectionselection and and breedingbreeding ??
Seed orchard• “Mommedeel” (° 1988 - 0.82 ha)
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
• 65 genotypes selected in two populations• Moderate yield ~ 20 kg in good crop years• Recent severe dieback
- unsuitable site- rejection of grafts
6
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
WhyWhy seedseed orchardsorchards ??
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
and and ……. . howhow ??
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Selection of constituents
Spatial design
Spatial isolation
Management
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
and and ……. . howhow ??
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Selection of constituents
SpatialSpatialSpatial designdesigndesign
SpatialSpatialSpatial isolationisolationisolation
ManagementManagementManagement
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SelectionSelection of of clonesclones
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SelectionSelection of of clonesclones
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SelectionSelection of of clonesclones
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SelectionSelection of of clonesclones
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Selection criteria• Disease resistance
- anthracnosis Blumeriella jaapii (observation of natural infections)
- bacterial canker PseudomonasPseudomonas syringaesyringae pvpv. . syringaesyringae / / morsprunorummorsprunorum / / aviiavii (artificial infections)
10
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
and and ……. . howhow ??
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
SelectionSelectionSelection of of of constituentsconstituentsconstituents
Spatial design
SpatialSpatialSpatial isolationisolationisolation
ManagementManagementManagement
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Neighbouring trees in future seed orchards should becross-compatible• phenologically: overlap of flowering period• gametophytically: different SI-genotype
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
(%)
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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)
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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 %
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Phenological compatibility• furthermore…
- no influence of exposure or location within the plantation
N
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Gametophytic compatibility – Molecular assessment• sizing of introns
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Gametophytic compatibility – Molecular assessment• sizing of introns• allele-specific PCR
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Gametophytic compatibility – Confirmation• controlled crosses
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial designdesign
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
and and ……. . howhow ??
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
SelectionSelectionSelection of of of constituentsconstituentsconstituents
SpatialSpatialSpatial designdesigndesign
Spatial isolation
ManagementManagementManagement
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
SpatialSpatial isolationisolation
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
Minimizing genetic pollution
21 % of the pollen is coming from outside the plantation
4%17%79%
400 m
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
and and ……. . howhow ??
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
SelectionSelectionSelection of of of constituentsconstituentsconstituents
SpatialSpatialSpatial designdesigndesign
SpatialSpatialSpatial isolationisolationisolation
Management
Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
Establishment & ManagementEstablishment & Management
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
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Seed Orchard Conference, 26 – 28 September 2007, Umeå, Sweden
StaffStaff
ClonalClonal SeedSeed OrchardOrchard of Wild of Wild CherryCherry –– ConstitutionConstitution and Designand Design
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
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