1. Cryogenic methods used to biodiversity conservation of common ash and three elm species in the Kostrzyca Forest Gene Bank Paulina Hrydziuszko Małgorzata Pałucka Czesław Kozioł 18th Cold Hardiness Seminar in Poland Institute of Dendrology, Polish Academy of Sciences Kórnik 14-15 May 2013

2. 2 "Conservation of biodiversity, through the conservation of ex situ common ash and three elm species in Lower Silesia in Natura 2000 areas"  Why did we choose ash and elm trees to protect biodiversity?  Ash dieback in Europe.  Elms dieback in Europe.  Examples of ash and elm protection in Poland.  Assumptions of our project.  Expected results. 18th Cold Hardiness Seminar Poland

3. 3 The Natura 2000 ares Forest Districts with habitats of elms Nature reserves of elms and ashes Forest Districts with habitats of common ash Range of the project covers an area of ​​ Lower Silesia. Species covered by the project have their positions on the surface protected areas (Natura 2000 sites, nature reserves). The location of common ash and three elm species in Lower Silesia in Natura 2000 areas.

4. 4 "Conservation of biodiversity, through the conservation of ex situ common ash and three elm species in Lower Silesia in Natura 2000 areas"  Dramatically rapid rate of extinction of the common ash from the 90’s of the XX th century and elms from the 20's and then in the 70's of XX th century, caused high concern to the survival of these species in many European countries.  The reasons of ash and elm trees dying were carefully diagnosed. In accordance to changes in the natural environment, such as river valleys conversion, riparian drainage, flood barriers constructions preventing flooding of the natural habitats, but the main cause are fungal pathogens.  Loss of biodiversity of elm-ash habitats was an inspiration for the Kostrzyca Forest Gene Bank to protect these species from extinction by implementation of the project entitled "Conservation of biodiversity, through the conservation of ex situ common ash and three elm species in Lower Silesia in Natura 2000 areas".  The main objective of the project is ex situ conservation of three species of elm and common ash through the cryogenic storage of seeds and shoots with dormant buds. The first part of the project will cover the terrain of Lower Silesia. 18th Cold Hardiness Seminar Poland

5. 5 Ash dieback in Europe - short history  1992 - first signs of severe dieback of ash in north-eastern Poland (Stocki 2001, Sierota et al. 1993)  After 1992 - the disease has been observed in other parts of Poland (Kowalski 2001, Przybył 2002) over an area of ​​ 10,000 hectares (Gil et al. 2006)  Ash dieback in Europe has been observed in Norway, Denmark, Lithuania, Austria, Belgium, Czech Republic, Finland, France, Germany, Hungary, Italy, Netherlands, Slovenia, Sweden, Estonia, Latvia and Switzerland  In 2012, the first signs of dieback are found in the British Isles (Coghlan 2012)  The speed of disease spreading according to Euforgen is 20-30 km per year (Euforgen, News, 14.12.2012). 18th Cold Hardiness Seminar Poland

6. 6  Austria - symptoms of F.excelsior L., F.angustifolia Vahl., F.ornus L. disease  Slovenia, Romania, Czech Republic, Germany - symptoms of F.excelsior L., F. angustifolia Vahl. disease [Schumacher et al. 2007, Jankovsky, Holdenrieder 2009]  Estonia - symptoms of F.excelsior L. F.mandshurica Rupr., F.nigra, F. pennsylvanica and F. americana L.  By the end of 2012, there was no ash dieback caused by Chalara fraxinea on any other continent outside Europe. Ash dieback in Europe - short history 18th Cold Hardiness Seminar Poland

7. 7 Subsequent years Changes in forest area of common ash as the dominant species [ha] in the years 2001-2011 based on data from the General Directorate of State Forests in Poland 18th Cold Hardiness Seminar Poland

8. 8 Discoloration of wood in the form of sectors and discoloration in the shape of the letter "T " Symptoms of ash dieback Cortical necrosis, wood cancers, local hypertrophy The fruiting bodies of the pathogen Hymenoscyphus pseudoalbidus (anamorphic stage-Chalara fraxinea (Kowalski 2006) Wilting, discoloration and local or full necrosis of leaves and their premature precipitation Dieback of whole branches or their peaks, irregular leaves of crown 18th Cold Hardiness Seminar Poland

9. 9 Symptoms of ash dieback 18th Cold Hardiness Seminar Poland

10. 10 Hymenoscyphus pseudoalbidus- Teleomorph stage of the pathogen. Pathogen's fruiting bodies form (VII-IX) at last year's ash leaf nerves, produce spores spread by wind. The spores infect the tree, starting the process of dying. Chalafa fraxinea- Pathogen's vegetative stage (anamorphic). Grows on infected fallen leaves. It produces a form of survival of the pathogen, which in the summer of next year will produce fruiting bodies Hymenoscyphus pseudoalbidus. 18th Cold Hardiness Seminar Poland

11. 11 Hymenoscyphus pseudoalbidus 18th Cold Hardiness Seminar Poland

12. 12 Genetic resources of Fraxinus excelsior L. in the Kostrzyca Forest Gene Bank Genetic resources of Fraxinus excelsior L. in the Kostrzyca Forest Gene Bank (01.03.2013 r.) SpeciesForest District Type and number of the seed base Mass [kg] Conditions of storage Fraxinus excelsior L. Szczecinek plus tree 3143 0,5600-10°C plus tree 3135 0,7508-10°C plus tree 3139 0,1578-10°C plus tree 4784 0,6662-10°C Szczecinekplus seed stand MP/2/31570/05 2,3100-10°C Mircze plus seed stand MP/2/31254/05 8,1360-10°C Opole Seed stand MP/1/5343/05 9,8900-196°C Summary22,4708 18th Cold Hardiness Seminar Poland

13. 13 DUTCH ELM DISEASE (DED)  Dutch elm disease (DED) is caused by a member of the sac fungi (Ascomycota) affecting elm trees and is spread by the elm bark beetle.  Although believed to be originally native to Asia, the disease has been accidentally introduced into America and Europe, where it has devastated native populations of elms, which had not had the opportunity to evolve resistance to the disease.  The factors impacts of DED are ascomycete microfungi. Three species are now recognized:  Ophiostoma ulmi, which afflicted Europe in 1910, reaching to North America by imported wood in 1928,  Ophiostoma himal-ulmi, a species endemic to the western Himalaya,  Ophiostoma novo-ulmi, an extremely virulent species which has been first described in Europe and North America in the 1940s and which devastated elms in both areas since the late 1960s. 18th Cold Hardiness Seminar Poland

14. 14 Symptoms of Dutch Elm Disease  DED symptoms are the result of the infection in the vascular system of the tree (system conducting water).  Infection causes clogging of vascular tissues and obstructing regular flow of water to the crown of the tree.  Symptoms are visible on the leaves, branches and stems. Final outcome of the disease is the death of the whole tree.  Symptoms are often observed in early summer, but they may be present at any time of the growing season. Early symptoms - still green but wilting leaves Summer - brown, wilted leaves Dieback of tree 18th Cold Hardiness Seminar Poland

15. 15 Changes in elm forest area Unfortunately, the Kostrzyca Forest Gene Bank does not have sufficient data on changes in the area occupied by different species of elms. In the State Forests National Forest Holding elms are treated only as a admixed species, not as the main tree species. 18th Cold Hardiness Seminar Poland

16. 16 Conservation of the ash and elms in Poland Protection recommendations (Chief Inspector for Environmental Protection) on the type of habitat: Riparian forests of oak and elm-ash (Code habitats 91F0) after monitoring research in the years 2009-2011 (monitoring of species and habitats with particular emphasis on Special Areas of Conservation Nature 2000)  Improving the hydrological regime of riparian forests (e.g. reconstruction of levees system-restore the effect of inundation)  Breeding activities, such as artificial replenishment of species which natural renewing is difficult (e.g. oaks along the Bug River)  Proposed modification procedure in stands areas where ash dominates and dieback is observed 18th Cold Hardiness Seminar Poland

17. 17 Conservation of the ash and elms in Poland To maintain habitat 91F0 it is necessary to:  Limit removal of ash trees, even at the cost of leaving some of them to the natural decay of the forest,  Extend the species composition in habitats  In areas where riparian forests have uniform-age oaks as dominant species, it is suggested to exclude some of them from forest management … What about the harvest of the seed?... 18th Cold Hardiness Seminar Poland

18. 18 Assumptions for the project which will be implemented in the Kostrzyca FGB 1.Verification of the number of individuals and their health at selected positions. 2.Collection of seeds and stems with buds ash resting for cryogenic storage. 3.Cryotherapy of common ash seeds to inactivate the pathogen. 4.Thermotherapy of common ash seeds to inactivate the pathogen. 5.Dissemination of the project results in the form of publications and brochures. 6.DNA isolation to prepare DNA data bank for the future molecular research. 18th Cold Hardiness Seminar Poland

19. 19  The project will be co-financed by the Regional Fund for Environmental Protection and Water Management in Wroclaw  Duration of the project covers the period: 2013 - 2016 Assumptions for the project which will be implemented in the Kostrzyca Forest Gene Bank 18th Cold Hardiness Seminar Poland

20. 20 The Kostrzyca Forest Gene Bank Miłków 300 58-535 Miłków 75 7131048 lbg@ lbg.lasy.gov.pl Thank you for your attention