Objective Priority Species Breeding Strategies Research ... · Objective Priority Species Breeding...

SelectionSelection and and BreedingBreeding ofofValuableValuable BroadleavedBroadleaved Tree SpeciesTree Species

ObjectiveObjective

PriorityPriority SpeciesSpecies

BreedingBreeding StrategiesStrategies

Research Research ToolsTools

SelectionSelection CriteriaCriteria


ObjectiveObjective

PriorityPriorityPriority SpeciesSpeciesSpecies

BreedingBreedingBreeding StrategiesStrategiesStrategies

Research Research Research ToolsToolsTools

SelectionSelectionSelection CriteriaCriteriaCriteria


ObjectiveObjectiveProducingProducing high high qualityquality forestforest reproductivereproductive materialmaterial throughthrough creationcreation and and / / oror selectionselection of of geneticallygenetically superior superior basicbasic materialmaterial withwith respect respect forfor the the maintenancemaintenance of a of a sufficientlysufficiently broadbroad geneticgenetic basisbasis

OrOr…….is .is therethere anyany otherother objectiveobjective forfor a a forestforest tree tree breederbreeder ??????


ObjectiveObjectiveObjective

PriorityPriority SpeciesSpecies





PriorityPriority tree speciestree speciesCriteriaCriteria

EconomicEconomic importanceimportance ((timbertimber productionproduction))ImplementationImplementation of of EUEU--directievdirectiev 1999/105/EC1999/105/ECDEMAND FRM >>SUPPLY FRMDEMAND FRM >>SUPPLY FRM

And And …… doingdoing ““a bit of a bit of everythingeverything”” unevitablyunevitably leadsleads toto““doingdoing somethingsomething prettypretty close close toto nothingnothing””

Wild Cherry (Prunus avium L.)

Common Ash (Fraxinus excelsior L.)


PriorityPriority tree speciestree species

Wild Cherry (Prunus avium L.)

Common Ash (Fraxinus excelsior L.)


PriorityPriority tree speciestree species

Very high prices (standing timber)

Aesthetically highly appreciated wood furniture industry

− Heavy assortments (DBH ~ 60 cm) 250 € / m³− Top quality 600 € / m³

SelectionSelection and and BreedingBreeding ofofWild Wild CherryCherry

Economic importance

Demand Supplymanagement plans of state and private forestsrequests for afforrestation grants for prtivate forests96.000 plants in 2004 = 50 kg seeds

Plants (x 1000) used forafforestation and stand-conversion in 2004

253

95

82413521

319 Inland oak

Wild cherry

Common ash

Beech

Black alder

Sycamore

Other

Plants used for afforestation and stand-conversion from 2001 till 2004

0102030405060708090

2001 2002 2003 2004

Plan

ts (x

100

0)

State forest Public forest Private forest Total


Demand Supply

Until 1 January 2003: “optional” tree species− Certificate of provenance non - compulsory− Certificate required for granting subsidies for afforestation− Sufficient supply of FRM, yet … quality ??

Since 1 January 2003: implementation of 1999/105/EC− Wild cherry provenances to be certified− High genetic quality and silvicultural value of FRM has to be

warranted


SUPPLY

Indigenous basic materialSeed orchard “Mommedeel”


Indigenous basic materialSeed orchard “Mommedeel”


− Forest of Meerdaal (Bierbeek)

− ° 1988− 0.85 ha− 65 clones originating from 2 native populations

(Solre-Saint-Géry & Anor)

− certfied in 2002 (B1502 AG)

−− unsufficientunsufficient productiveproductive capacitycapacity (~ 20 kg)(~ 20 kg)

−− sporadicsporadic seedseed yearsyears mastjaren mastjaren (1999 (1999 –– 2003)2003)

−− recente recente diedie--backback phenomenaphenomena ((incompatibilityincompatibility rootstock rootstock –– graftgraft))

Seed Stand “Vrebos”

− “high” sales price purchase of foreign provenances− introgression from sweet cherry cultivars suspected

− Kortenberg− 0.28 ha− certified in 2002 (B1005 AG)

− unsufficient productive capacity (~ 15 kg)

− sporadic seed years (2002)

− not cost-effective: cost price = 4 x sales price

Indigenous basic material

− genetic diversity ??


Seed Stand “Vrebos” – Genetic diversity ??

Indigenous basic material


− diversity assessed with 13 SSR

− 321 individuals− 189 different genotypes− 28 clonal clusters consisting of 49.9 % of the individuals− trees per cluster: 2 42 − area of cluster: 0.10 302.35 m²− distance between tree: 0.10 36.98 m− heterozygosity


0 50 100 150 200x

-100

-50

0

50

y

0 50 100 150 200x

-100

-50

0

50

y


Seed Stand “Vrebos”Indigenous basic material


− to be investigated• age structure of stand• parenthood analysis• cross-compatibility of genotypes

phenological compatibility (flowering period)gametophytic (in)compatibility (SI-system)




Breeding Strategies




BreedingBreeding StrategyStrategyBreedingBreeding isisDoingDoing youryour bestbestSelectionSelection the bestthe bestAndAnd……HopingHoping forfor the bestthe best

However

Provenance trials List of Recommended Provenances

Selection of Seed Stands

Creation of Seed Orchards





Research Tools



ResearhResearh ToolsTools

Field trials (clonal - progeny - provenance trails)

Quantitative genetics relevance of selection criteria

Fytopathological research (resistance breeding)

Molecular support






Selection Criteria


BREEDING = SELECTINGBREEDING = SELECTING

SELECTING = CRITERIASELECTING = CRITERIA

Vigour

Morphology

Phenology

Wood Quality

Disease resistance

Genetic diversity


StrategyStrategy 1 1 –– Provenance Researchestablishment of 1 provenance trial in 2002

collaboration INBO & CRNBF (B) – LOBF (D) – Alterra (NL)

small-scale: 11 provenances (B:4 – NL:3 – D:4)

identification of provenances at the seed stand and seed orchard level

COST E42: forum for exchange of FRM (cf. RAP) ??


StrategyStrategy 2 2 –– Seed Stands

few perspectives for selection of aditional “natural” seedstands

alternative strategy: “creation” of seed stands

Certified Basic Material Selected plus trees

Seed harvest

Selection seedlingsin nursery

Non-certifief BM

Experimentalplantation

Seed harvest

Selectionplus treesSeed orchard


Mommedeel

Creation of a new generation clonal seed orchard− higher productive capacity− higher genetic quality of off-spring (FRM)

Three major research topics− selection of mother trees− spatial design− spatial isolation

Practical realisation

StrategyStrategy 3 3 –– Seed Orchards


A A newnew seedseed orchardorchard –– Selection of mother treesSource = basic collection of van 159 genotypes

114 genotypes selected 11 populations (1976 – 1986)

Selection matureplus trees

Vegetativepropagation

Selection seedlingsin nursery

Seed harvestplus trees

Experimentalplantations

Selection juvenileclones

6 multi clonal plantations

Location Planting Nbr. Areaclones (ha)

Herne 1983 38 0.27 Grimminge I 1979 7 0.12 Grimminge II 1980 4 0.42 Grimminge III 1986 98 0.19 Moustier 1984 48 0.55 Mommedeel 1988 65 0.85

45 genotypes selected in 16 populations (1998 – 1999)


A A newnew seedseed orchardorchard –– SelectionSelection of of mothermother treestreesSelectie strategy (1/3)

Identification of identical genotypes (AFLP & 13 SSR)168 selected plus trees 159 different genotypes

Criteria:− vigour (tree height)

− form (stem straightness, branching habit, apical dominance)

− phenologie (flushing, bud set, flowering)

− disease resistance (bacterial canker, anthracnosis)

Clonal trials− in vitro propagation of all genotypes− establishment of 8 experimantal plots ( 5.4 ha )− estamition of heritability h²G of selection criteria


Afstammingstesten− seed collection in multiclonal plantations half-sib progeny− establishment of 13 experimental plantations ( 11.3 ha )− estimation of heritability h²A of selection criteria− assessment of General Combining Ability (GCA) for each trait

for each genotype

n m i n m iGCA i = ( Σ Σ Tij ) / ( Σ m i ) - ( Σ Ti j ) / m ii=1 j=1 i=1 j=1

with m i = nbr. of seedlings within the hal-sib progeny of genotype in = number of genotypes

Selectie strategie (2/3)A A newnew seedseed orchardorchard –– SelectionSelection of of mothermother treestrees


Construction of a selection-index Isn

Is = Σ ( h²A * GCA ) ii = 1

with n = number of traits assessed

Selection of genotypes with Is – value > median

Selectie strategie (3/3)

A A newnew seedseed orchardorchard –– SelectionSelection of of mothermother treestrees


Patch-like pollination pattern in wild cherry populations ?test-case: seed orchard Mommedeelparenthood analysis− selection of 16 equally distributed mother trees− ad random selection of 30 seedlings in the half-sib offspring of

each mother tree− identification of fathers using 13 SSRs and the SI-genotype

leave a scent mark on flowers they visitforage on small clusters of trees

Wild cherry = entomophilous

Pollen vector = bumble bees (Bombus spp.)


A A newnew seedseed orchardorchard –– SpatialSpatial designdesign



Locus No. Alleles Size range HO HE DP PceGA34 13 129 - 165 0.89 0.86 0.95 Ps09F08 6 139 - 159 0.67 0.69 0.88 EMPA014 8 216 - 230 0.59 0.55 0.77 EMPaS01 4 222 - 238 0.75 0.63 0.72 EMPaS10 6 151 - 183 0.94 0.74 0.83 EMPaS11 4 73 - 106 0.69 0.54 0.63 EMPaS12 5 122 - 148 0.75 0.70 0.83 EMPaS14 3 196 - 210 0.75 0.66 0.75 EMPaS02 5 133 - 145 0.81 0.75 0.87 EMPaS06 5 200 - 218 0.69 0.71 0.81 EMPA004 4 180 - 188 0.79 0.71 0.79 EMPA005 4 237 - 253 0.50 0.52 0.71 EMPA015 7 216 - 239 0.71 0.76 0.88 EMPA018 5 94 - 106 0.57 0.60 0.79 SI 19 1,00 0,96

Multiplex AMultiplex B

Internal pollenflux38 % of father within the seed orchard

75 % of pollen donors within a distance of 10 m frommother tree (= 2 x planting distance)

0

5

10

15

20

25

30

35

40

45

Con

trib

utio

n to

pol

linis

atio

n (%

)

5 10 15 20 25 30 35 40

Distance from mother tree (m)



Spatial arrangement of mother trees in futur seed orchard: neighbouring trees should

be cross-compatible

• phenologically: overlap of flowering period• gametophytically: different SI-genotype

produce an offspring superior compared to any otherrandom father / mother combination (high SCA)



Phenological compatibility

observation of flowering period

• crown divided in 4 strata



• observation of share of fully opened flowers per stratum everytwo days from 02/04/2005 10/052005

• 16560 observations !


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

(%)


however…• fully opened flowers only receptive for 38 hours• viability of pollen in pollen bags of bumble bees = 12 hrs



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 since f irst o bservat io n (ho urs)


no significant difference between ramets of one genotype

observations of 2002 and 2004 are concordant

no influence of exposure and location of tree withinthe seed orchard



N





1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1721 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 4142 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 6162 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 8283 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103

104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393

Sx Sy

S-locus

Gametophytic self-incompatibilitygenetic basis: S – allelesS – locus = polymorph: 13 S - alleles in cv’ s + 6 in wild cherry



S1S2 S1S2 S1S2

S3 S4 S3 S1 S1 S2

Compatible Semi-Compatible Incompatible


Gametophytic self-incompatibilitygenetic basis: S – allelesS – locus = polymorph: 13 S - alleles in cv’ s + 6 in wild cherrygenotypes with the same SI-genotype are cross-incompatible



Gametophytic self-incompatibility

molecular identification of S-alleles

sizing of introns


2nd Intron 1st Intron



molecular identification of S-alleles

sizing of introns

allel-specific PCR




control through controlled crosses




sequencing of new alleles



Specific Combining Ability


Genotype G1 G2 G3 G4 G5 G6 G7 … Gn-1 Gn

G1 T12 T13 T14 T15 T16 T17 T1(n-1) T1n

G2 T23 T24 T25 T26 T27 T2(n-1) T2n

G3 T34 T35 T36 T37 T3(n-1) T3n

G4 T45 T46 T47 T4(n-1) T4n

G5 T56 T57 T5(n-1) T5n

G6 T67 T6(n-1) T6n

G7 T7(n-1) T7n…

Gn-1 T(n-1)(n)

Gn

DI-ALLEL CROSSES

Minimizing genetic pollution

Mommedeel: 62 % of the pollen is coming from outside the orchard

From where….?

A A newnew seedseed orchardorchard –– SpatialSpatial isolationisolation




Minimizing genetic pollution

Mommedeel: 62 % of the pollen is coming from outside the orchard

From where….?



38%14%44%4%

A A newnew seedseed orchardorchard –– PrakticalPraktical realisationrealisation

Vegetative propagation of selected mothertreesgrafting on dwarfing rootstock“Gisela 5”: compromis between dwarfing / compatibility

Planting


A A newnew seedseed orchardorchard –– Practical Practical realisationrealisation

Vegetative propagation of selected mothertreesgrafting on dwarfing rootstock“Gisela 5”: compromis between dwarfing / compatibility

Planting


A A newnew seedseed orchardorchard –– Practical Practical realisationrealisation

Managementpruningcontrol of herbaceous vegetationbird netsbumble bee hives


ThankThank youyou forfor youryourkind kind attentionattention

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