Diversity in European Chestnut (Castanea sativa Mill.

Emily Buck

Talk Overview
Introduce European Chestnut Biology History

Economic Value
Concerns for the Species Future CASCADE - EU project CASCADE at East Malling Preliminary Results Conclusions and Discussion

Biology - Castanea sativa Mill.

Classification Family: Fagacea Genus: Castanea 13 Castanea species C. sativa, indigenous to Europe

Monoecious
Wind (insects) pollinated Age and height 30-35m, 150+ years old Tree of 100 Horses 2000-4000 yrs

History
Spread of C. sativa pre - glaciation

Centre of origin

History
Spread of C. sativa post - glaciation

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
Global Average Temperature 14.4

seasonal variation
disease spread Phytophthora (ink disease) Cryphonectria (chestnut blight) Human intervention selection management practice Implication of habitat loss

13.5 1860

Year

2000

CASCADE - Partners
Dr Villani (co-ordinator), CNR-IAS Dr Vannini, University of Tuscia Dr Botta, University of Torino Dr Romane, CNRS-CEFE Dr Robin, INRA Dr Fernandez-Lopez, CIFL-Lourizan

Dr Eriksson, University of Agricultural Science

Dr Aravanoupoulos, Aristotele University of Thessaloniki Dr Diamandis, NAGREF Miss Russell, HRI East Malling Dr Garrod, University of Newcastle

CASCADE

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

CASCADE
Main areas of research Distribution, ecology and management plant surveys, site history questionnaire

Gene dispersal and genetic make-up application of molecular markers East Malling involved Variation in adaptive traits QTL mapping drought, frost resistance field performance East Malling involved

CASCADE
Main areas of research

4 Variation in disease resistance

incidence of Phytophthora sp. resistance; inoculate seedlings, shoots

5 Socio-economic & environmental impact

questionnaires growers and consumers

6 Strategy for integrated conservation and

utilisation integration research areas 1-5

CASCADE
Populations: Natural stand Coppice Orchard Extreme sites Other sites

CASCADE 3 Population Types

Natural stand

Coppice

Orchard

Simple Sequence Repeats (SSRs) Microsatellites

..AGAGAGAGAGAGAG..
33P

labelled primers autoradiography

Fluorescent primers ABI 3100 prism

Italy and East Malling - SSR Enriched libraries C. sativa

Results - SSR primer Development

13 East Malling Castanea sativa (EMCs) SSR primers
Primer EMCs2 EMCs4 EMCs10 EMCs11 EMCs13 EMCs14 EMCs15 EMCs17 EMCs22 EMCs25 EMCs32 EMCs38 EMCs42 Repeat motif Number of putative alleles 3 3 3 2 3 1 5 4 6 5 8 12 3

[CGG]7 [GGC]7 [CA]8 [GGA]7[GGC]6 [GCA]8 [GAG]7 [CAC]9 [AGC]4[CCAA]5 [GA]19 [GA]12 [AG]18 [AG]31 [CA]11

Results - CASCADE SSR Mapping

1 2 3 4

10

11

12

UK Italian

12 linkage groups (chromosomes), 829 cM 458 markers: RAPDs, ISSR, Isoenzymes, AFLP, STS, SSRs

Results - SSRs for Diversity Screen

1 2 3 4

10

11

12

UK Italian

Not mapped

Screening diversity: 8 mapped SSRs, 3 not mapped SSRs

CASCADE - Diversity Screen

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

Results East Malling Diversity Screen

6 polymorphic SSR loci
Linkage Group 7 9 4 1 Number of Alleles At present Initially (20) 5 3 6 5 12 5 25 12 11 6 22 8

Primer EMCs4 EMCs15 EMCs25 EMCs38 CsCAT4 CsCAT6

Total of 81 alleles found so far

Results
Preliminary relationships between individuals:
Natural populations UK populations

Spain UK France Italy Greece

Results
Spain UK France Italy Greece Natural populations & UK

Central Greece

Greece populations discrete groups

Individuals from Central and Northern Greece in separate groups Northern Greece

Results
Spain UK France Italy Greece Natural populations & UK Spain

Northern and Southern Spain Group Southern Italy group, from Sicily
UK and French generally dispersed throughout

Southern Italy

Conclusions and Discussion

Natural
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?

Conclusions and Discussion

Natural
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

Conclusions and Discussion

Orchard and Coppice Diversity within Orchards and Coppice populations varies region dependant may be result of different selection pressure and history of given population? Diversity in UK populations unusual diverse for marginal population multiple imports from different areas at different times? But very susceptible to Phytophthora (Ink disease) Additional data from other markers

Acknowledgements
Karen Russell David Blakesley Christine Cahalan Dave Moodley, Suad AlMarrooei, Bruce Sutherland, Dan Sargent, Bobbie Maxted, everyone in the lab PhD advisor: PhD supervisors:

CASCADE Project co-ordinator: Fiorella Villani Italy: R. Botta, D. Marioni, M. Casasoli Greece: P. Aravanoupoulos, S. Diamandis Spain: J. Fernandez-Lopez France: F. Romane, C. Robin, A. Grossmann, T. Barreneche

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