Genetics and Population structure on an English yew gene conservation forest at foothills of the Eastern Alpine Mountains

1. 17/04/10 GENETICS AND POPULATION STRUCTURE OF AN ENGLISH YEW GENE CONSERVATION FOREST AT FOOTHILLS OF THE EASTERN ALPINE MOUNTAINS Raphael KLUMPP, AMALESH DHAR, Bernhard AIGNER, Herwig RUPRECHT, Harald VACIK

4. World Distribution 17/04/10 The circumpolar distribution of Taxus baccata and other Taxus species from Hultén & Fries (1986). + indicates interglacial records; indicates isolated occurrences; hatched areas indicate regions of common or fairly common occurrence. Broken lines outline the distribution of Taxus species other than T. baccata . Thomas P.A., Polwart A. 2003. Taxus baccata L. biological flora of the British Isles 229. Journal Ecology 91: 489–524 T.baccata T.brevifolia T.canadensis T.floridana T.globosa T.cuspidata T.chinensis T. wallichiana

5. Taxus baccata - Introduction 17/04/10 Introduction - Yew is one of the most primeval tree species in Europe, originated 2 million years ago in the late Tertiary period - It is a slow-growing, long-living, shade loving evergreen conifer tree species in temperate forests - At present it has gained considerable attention to the researchers as a source of anti-cancer drug Texon - Yew is also a rare endangered and prone of extinction tree species in Austria

7. Research Objectives 17/04/10 The main objectives of this study : 2. To estimate the level of genetic variation and diversity of selected Austrian yew population 1. To characterize ecological parameters for the growth and development of an Yew population in Austria 3. To propose the possible conservation measures

8. Materials and Method 17/04/10 Materials and Method

11. Materials and method 17/04/10 Classification scheme for the vitality of Taxus baccata 1 = Universally strong crown, 2 = weakly developed, constricted crown, 3 = undeveloped, most unilaterally, clamped crown, 4 = almost dying crown (for detail Dhar et al 2006) Vitality class Attributes Percentages of Living crown Foliage density Crown formations A (Very vital) > 70 % > 90 % 1 or 2 B (Vital) 50 - 70 % 75 - 90 % 1 or 2 C (Less Vital) 30 - 50 % < 75 % 1, 2 or 3 D (The least vital) < 30 % < 75 % 1, 2 ,3 or 4

12. Materials and Methods – Structural Indices 17/04/10 Quantifying the Structural Diversity Three neighbouring trees of each individual male and female yew at each sample plot has been used to calculate following structural indices ( Füldner1995) 35cm, 22m 20cm, 18m 22cm, 11m 2 nd Neighbour 3,5m 60cm, 32m 3 Neighbour Target tree yew 1 st Neighbour 1,5m 4,7m Attributes of stand Structure Calculated formula Interpretation Mingling Distance to Neighbour DBH differentiation Height Differentiation

13. Materials and Method- Genetics 17/04/10 Genetic Structure: Sample: trees buds Sample size: 95 Biochemical marker: Isozyme Enzyme system: 7 laboratory procedure according to: Hertel 1996 Data evaluation: GSED (Gillet 1998) Following genetic parameters were assessed Polymorphic Loci (95%): (0.95 criterion: a locus is considered polymorphic if the frequency of the most common allele does not exceed 0.95, P >95 % ) A/L - average number of alleles per locus He - average expected heterozygosity= the estimated fraction of all individuals who would be heterozygous for any randomly chosen locus Ho- observed heterozygosity = No.of Heterozygotes at a locus / total no of investigated individuals hypothetical gametic diversity: the potential of a population for producing genetically diverse gametes

14. Materials and Method - Genetics 17/04/10 Enzyme system s used for electrophoretic analysis of the yew population Enzyme Gene locus Allele no E. C. number Aspartate-aminotransferase AAT- A AAT- B 2 3 2.6.1.1 Isocitrate-dehydrogenase IDH-A IDH-B 3 4 1.1.1.42 Leucine-aminopeptidase LAP-A LAP-B 4 5 3.4.11.1 Phophoglucose isomerase PGI-B 5 5.3.1.9 Phosphoglucomutase PGM - A 4 2.7.5.1 6-Phosphogluconate-dehydrogenase 6-PGDH- A 2 1.1.1.44 Shikimate-dehydrogenase SKDH-A 3 1.1.1.25

15. Results and Discussion 17/04/10 Results and Discussion

16. Results and Discussion- Population Structure 17/04/10 Stand characteristics of T. baccata population Species composition of Stiwoll gene conservation forest Fagus sylvatica Abies alba Taxus baccata Picea abies and other broad leaf species Stand Area [ha] Other trees species DBH ≥ 5.0 cm [n ha -1 ] Tree basal area [m 2 ha -1 ] Tree volume [m 3 ha -1 ] No of Yew DBH ≥ 5.0 cm [n ha -1 ] Study site 2.6 661 (+/- 20 %) 36.7 384 97

17. Results and Discussion - Population Structure 17/04/10 Number fo individuals of T. baccata in the netural regeneration according to height classes Vitality classes of T. baccata according to diameter distribution Vitality class distribution of T. baccata population

18. Results and Discussion: Structural Diversity 17/04/10 Mingling distribution (DMi) for English yew in Mondseee [0,00 - all 3 yews, 0,33 - two yews, 0,67 - one yew, 1,00 - no yew as surrounding neighbour to the reference tree]

19. Results and Discussion: Structural Diversity 17/04/10 Dristibution of distance to nearest neighbours (Dj) for English yew in Mond See

20. Results and Discussion: Structural Diversity 17/04/10 Diameter differentiation with English yew and it neighbours negative differentiation Positive differentiation Larg middle small middle Large Very larg small Very Large

21. Results and Discussion: Structural Diversity 17/04/10 Positive differentiation Height differentiation with English yew and its neighbours Larg middle small middle Large Very larg small negative differentiation

22. Results and Discussion 17/04/10 Genetic structure

23. Results and Discussion - Genetics 17/04/10 Comparing genetic parameters for different studies on Taxus baccata in Europe Species Sample size No of gene loci Parameters Hypo. gametic diversity Different studies P 95 (%) A/L Ne Ho He Taxus baccata 95 9 78 2.44 1.43 0.257 0.304 42.60 This study 30 18 61.11 2.83 1.37 0.286 0.279 -- Lewandowski et al. (1995) 9-58 6 80.6 2.62 1.48 0.340 0.316 -- Cao et at. (2003) 50-80 5 -- -- 1.4 0.302 0.308 7.84 Tröber et al. (2004)

26. Acknowledgements 17/04/10 Acknowledgements For financial support Landesregierung Steiermark ÖAD for North South Dialogue Scholarship ÖOG for One world Scholarship

27. 17/04/10 A malesh DHAR Institut of Silviculture Department for Forest-and Soil Sciences University of Natural Resources ans Applied Life Sciences Peter Jordan-Str. 82, A-1190 Wien Tel.: +43 1 47654-4075, Fax: +43 1 47654-4092 Email: amalesh.dhar@boku.ac.at ,Web: www.boku.ac.at Thanks for kind attention