Diversity in European Chestnut (Castanea sativa Mill.)

Emily Buck

Talk Overview

• Introduce European Chestnut – Biology

– History

– Economic Value

– Concerns for the Species Future

• Conclusions and Discussion

• Preliminary Results

• CASCADE - EU project

• CASCADE at East Malling

Biology - Castanea sativa Mill.

• Classification – Family: Fagacea

– Genus: Castanea– 13 Castanea species– C. sativa, indigenous to Europe

• Age and height

– 30-35m, 150+ years old

– ‘Tree of 100 Horses’ - 2000-4000 yrs

• Monoecious

• Wind (insects) pollinated

History

Centre of

origin

Spread of C. sativa pre - glaciation

Spread of C. sativa post - glaciation

History

Economic Value

• Castanea sp: Nuts

– World production: 510,702 t/yr

– Asian 64%, Europe 30%

– C.sativa & hybrids Turkey: 70,000 t/yr

• Castanea sativa: Wood

– Mainland Europe:

structural and agricultural

– UK: agricultural use; fences

– 18, 788 ha in UK; 42% coppice

– Coppice: stems cut 3-25 year rotation

Concerns for the Species Future

• Climate change

– water and temperature

– seasonal variation

– disease spread Phytophthora (ink disease)Cryphonectria (chestnut blight)

– selection– management practice

Global Average Temperature

ºC

14.4

13.51860 2000

Year

• Human intervention

• Implication of habitat loss

Dr Romane, CNRS-CEFEDr Robin, INRA

Dr Villani (co-ordinator), CNR-IASDr Vannini, University of TusciaDr Botta, University of Torino

Miss Russell, HRI East MallingDr Garrod, University of Newcastle

Dr Eriksson, University of Agricultural Science

Dr Fernandez-Lopez, CIFL-Lourizan

Dr Aravanoupoulos, Aristotele University of Thessaloniki

Dr Diamandis, NAGREF

CASCADE - Partners

Securing Gene Conservation, Adaptive, Breeding Potential and Utilization of a Model Multipurpose tree species (Castanea sativa Mill.) in a Dynamic Environment

CASCADE

• 5th EU Framework Programme• Work programme 2: Energy, Environment and Sustainable

Development.• Key Action: Global Change, Climate and Biodiversity. • Thematic Priority: Assessing and Preserving Biodiversity.

http://soi.cnr.it/~chestnut/

1 Distribution, ecology and management– plant surveys, site history questionnaire

2 Gene dispersal and genetic make-up– application of molecular markers– East Malling involved

3 Variation in adaptive traits – QTL mapping – drought, frost resistance– field performance – East Malling involved

Main areas of research

CASCADE

5 Socio-economic & environmental impact– questionnaires growers and consumers

6 Strategy for integrated conservation and utilisation– integration research areas 1-5

Main areas of research

CASCADE

4 Variation in disease resistance– incidence of Phytophthora sp.– resistance; inoculate seedlings, shoots

CASCADE

Populations: Natural stand Coppice Orchard

Extreme sites

Other sites

CASCADE - 3 Population Types

Natural stand

Coppice Orchard

Fluorescent primers ABI 3100 prism

Simple Sequence Repeats (SSRs) - Microsatellites

33P labelled primers autoradiography

………..AGAGAGAGAGAGAG………..

• Italy and East Malling - SSR Enriched libraries C. sativa

Primer Repeatmotif

Number ofputative alleles

EMCs2 [CGG]7 3

EMCs4 [GGC]7 3

EMCs10 [CA]8 3

EMCs11 [GGA]7[GGC]6 2

EMCs13 [GCA]8 3

EMCs14 [GAG]7 1

EMCs15 [CAC]9 5

EMCs17 [AGC]4[CCAA]5 4

EMCs22 [GA]19 6

EMCs25 [GA]12 5

EMCs32 [AG]18 8

EMCs38 [AG]31 12

EMCs42 [CA]11 3

Results - SSR primer Development 13 East Malling Castanea sativa (EMCs) SSR primers

1 32 54 6 87 109 1211

Results - CASCADE SSR Mapping

• 12 linkage groups (chromosomes), 829 cM

• 458 markers: RAPDs, ISSR, Isoenzymes, AFLP, STS, SSRs

UK Italian

1 32 54 6 87 109 1211

Results - SSRs for Diversity Screen

• Screening diversity: 8 mapped SSRs, 3 not mapped SSRs

Not mappedUK Italian

• Application of polymorphic SSRs

– Total of 11: 6 East Malling, UK

5 University of Torino, Italy

• Screen CASCADE environmentally extreme sites

– Natural, coppice and orchard populations

– 8 trees and 20 seeds from each population

– Total of 3,864 samples

CASCADE - Diversity Screen

Results - East Malling Diversity Screen

Number of AllelesPrimer

LinkageGroup Initially (20) At present

EMCs4 7 3 5EMCs15 9 5 6EMCs25 - 5 12EMCs38 4 12 25CsCAT4 - 6 11

CsCAT6 1 8 22

• 6 polymorphic SSR loci

• Total of 81 alleles found so far

Results

Preliminary relationships between individuals:

– Natural populations

– UK populations

Spain

Greece

FranceUK

Italy

Central Greece

Results Natural populations & UK

Northern Greece

• Greece populations discrete groups

• Individuals from Central and Northern Greece in separate groups

Spain

Greece

FranceUK

Italy

Southern Italy

Results Spain

Greece

FranceUK

Italy

• Southern Italy group, from Sicily

• UK and French generally dispersed throughout

Spain • Northern and Southern Spain Group

Natural populations & UK

Conclusions and Discussion

• Two distinct groups from Northern and Central Greece

– diversity within Greek populations low

– variation in adaptability experiments:

Greek samples low adaptability

– unique group?, susceptible to climate change?

• Distinct group from Southern Italy, dispersed individuals from Northern Italy

– diversity within Southern Italy population low

– diversity within Northern Italy population higher similarities samples in Spain, France and UK

– Romans spread from this region?

Natural

Conclusions and Discussion

• Group of Spanish individuals

– diversity within Spain populations low?

– some samples dispersed, spread from refugia?

• French samples dispersed

– natural spread / introduced from diverse Italian site, or

Spanish refugia

Natural

Conclusions and Discussion

• Diversity within Orchards and Coppice populations varies– region dependant – may be result of different selection pressure and

history of given population?

Orchard and Coppice

Additional data from other markers

• Diversity in UK populations– unusual diverse for marginal population – multiple imports from different areas at different times?

But very susceptible to Phytophthora (Ink disease)

Acknowledgements

CASCADEProject co-ordinator: Fiorella VillaniItaly: R. Botta, D. Marioni, M. CasasoliGreece: P. Aravanoupoulos, S. DiamandisSpain: J. Fernandez-LopezFrance: F. Romane, C. Robin, A. Grossmann,

T. Barreneche

http://soi.cnr.it/~chestnut/

PhD advisor: Karen RussellPhD supervisors: David Blakesley

Christine CahalanDave Moodley, Suad AlMarrooei, Bruce Sutherland,

Dan Sargent, Bobbie Maxted, everyone in the lab