Diversity in European Chestnut ( Castanea sativa Mill.)

1. Diversity in European Chestnut (Castanea sativa Mill.) Emily Buck

2. CASCADE - EU project • CASCADE at East Malling Talk Overview • Introduce European Chestnut • Biology • History • Economic Value • Concerns for the Species Future • Preliminary Results • Conclusions and Discussion

3. 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

4. History Spread of C. sativa pre - glaciation Centre of origin

5. History Spread of C. sativa post - glaciation

6. 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

7. Global Average Temperature 14.4 ºC 13.5 1860 2000 Year • Implication of habitat loss Concerns for the Species Future • water and temperature • seasonal variation • disease spread Phytophthora (ink disease) Cryphonectria (chestnut blight) • Climate change • Human intervention • selection • management practice

8. 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

9. 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 andSustainable Development. • Key Action: Global Change, Climateand Biodiversity. • Thematic Priority: Assessing and PreservingBiodiversity. http://soi.cnr.it/~chestnut/

10. 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

11. CASCADE Main areas of research • Variation in disease resistance • incidence of Phytophthora sp. • resistance; inoculate seedlings, shoots • Socio-economic & environmental impact • questionnaires growers and consumers • Strategy for integrated conservation and utilisation • integration research areas 1-5

12. Populations: Natural stand Coppice Orchard Extreme sites Other sites CASCADE

13. CASCADE - 3 Population Types Natural stand Orchard Coppice

14. Simple Sequence Repeats (SSRs) - Microsatellites ………..AGAGAGAGAGAGAG……….. 33P labelled primers autoradiography Fluorescent primers ABI 3100 prism • Italy and East Malling - SSR Enriched libraries C. sativa

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

16. 1 2 4 7 3 5 6 12 8 9 11 10 UK Italian Results - CASCADE SSR Mapping • 12 linkage groups (chromosomes), 829 cM • 458 markers: RAPDs, ISSR, Isoenzymes, AFLP, STS, SSRs

17. 1 2 4 7 3 5 6 12 8 9 11 10 UK Not mapped Italian Results - SSRs for Diversity Screen • Screening diversity: 8 mapped SSRs, 3 not mapped SSRs

18. 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

19. Results - East Malling Diversity Screen • 6 polymorphic SSR loci • Total of 81 alleles found so far

20. Spain UK France Italy Greece Results Preliminary relationships between individuals: • Natural populations • UK populations

21. Spain UK France Italy Greece Results Natural populations & UK • Greece populations discrete groups • Individuals from Central and Northern Greece in separate groups Central Greece Northern Greece

22. Spain UK France Italy Greece Results Natural populations & UK Spain • Northern and Southern Spain Group • Southern Italy group, from Sicily • UK and French generally dispersed throughout Southern Italy

23. 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?

24. 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

25. 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

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