Genetic improvement of ash ( Fraxinus excelsior ) a European project. RAP : Realising Ash s Potential. Dr. Gerry Douglas

Transcription

1 Genetic improvement of ash ( Fraxinus excelsior ) a European project RAP : Realising Ash s Potential Dr. Gerry Douglas Teagasc, Agriculture and Food Development Authority Kinsealy Research Centre, Malahide Rd. Dublin 17, Ireland

2 3 Workpackages Biodiversity assays Propagation technology Utilisation strategies

3 OVERALL OBJECTIVES: 1. To characterise the genetic diversity on regional & European scale 2. To accelerate the utilisation of scarce selections by vegetative propagation and induced flowering 3. To identify effective ways to exploit & use results & materials

4 Contracting partners & contacts Dr. Gerry Douglas Teagasc, IRELAND Dr. Jean Dufour INRA Dr. A. Meier Dinkel NFVA Dr. D. Thompson Coillte Teo., Dr. J. Hubert Forest Research, UK Dr. N.Frascaria-Lacoste, ENGREF, Dr. B. Heinze FFRC, AUSTRIA,... Dr J.F. Hausman CRP-GL, LUXEMBOURG... Mr. L. Somer Vitroform, Dr. M. Capuana IMGPF CNR, ITALY... Mr. B. De Cuyper INBO, Ms. K. Russell EMR, Dr. P. Mertens MRW BELGIUM. Dr. Andreas Ottitsch EFI, FINLAND..

5 Biodiversity assays WP 1: Biodiversity assays Growth & adaptation analyses Populations Provenance trials (10-15 yr old) Families Molecular analyses Genetic diversity Gene flow Hybridisation frequency New International Ash provenance trial Evaluation of existing provenance trials of ash Fraxinus excelsior

6 EVALUATION OF 52 ASH PROVENANCES (D Jacques, Gembloux B.) Genotypic Heritability Character Significance Site 1 Site 2 Height & Circumference ** Stem & Crown Form Forking ** NS Frost Damage ** Game Damage NS - -

7 Global analysis of provenance trials ( 3 countries 50 provenances each in common France Belgium Germany) Coefficient of variation as a % of the provenances, site and interaction effects ( P. Mertens Gembloux) Coef. Varn. % Stem girth Stem height Stem form Provenance Site Interaction Residual Site and Interaction effects > provenance effect. 2. Stem form has a relatively higher provenance effect relative to site and interaction effects. ** Efficiency of provenance selection is not yet demonstrated. 3. Progeny selection is not more useful than Provenance selection

8 Summary results Heritability values for ash: Very high for----susceptibility to frost -- bud flushing date (At the level of country, provenance and progeny) High for Stem straightness Height and circumference Global analysis of existing trials ----Very strong site effects and interaction effects --- stemform a good selection criterion Much intra population variation clonal selection

9 Biodiversity assays New European Provenance Trial 600 Seedlots collected & germinated from 44 EU provenances Testing 30 provenances core collection in 6 countries -- Material already planted in France. Germany Belgium, Italy, Ireland & Denmark

10 Ash provenences in the new European trial

11 Nursery flushing of 40 provenances on seven European sites, (F, D, IRL, UK, IT, B) first results ( B. De Cuyper) Provenance Country source Flushing Pattern North and Western IRL, UK, DK, LT Late (Stable) South + East I, CZ Early (Stable) Central B, F, D Intermediate LT + F Lt (Kaisiadorys) F (Sila Grande) Early Late

12 Biodiversity assays Chloroplast DNA markers ( maternal inheritance) ( sequenced variable regions ) atpb-rbc Mat-K Measuring genetic variation of ash Molecular Nuclear DNA markers Microsatellites ( five variable loci) FEMSATL 11, 9, 4, 16, M2-30 AFLPs Analysis of provenances, populations, clonal material

13 Biodiversity : Molecular analyses : Some results ( N. Frascaria Lacoste, B. Heinze, J. Fernandez, G. Douglas) Genetic Diversity estimates High for intra population diversity heterozygote deficiency Gene flow estimates are % of pollen from outside 12ha stand Gene flow ( seed & pollen flow within stands is restricted )

14 Genetic variation of Ash in Ireland M. Harbourne T. Hodkinson G. Douglas Natural populations 14 Provenances Seed orchard trees

15 Allele number and overall gene diversity in Irish ash using nuclear microsatellites Provenances 14 n=6 / provenance Natural populations 3 n=50 / pop Seed orchard Ms- Locus No. Alleles Gene diversity H T No. Alleles Gene diversity H T No. Alleles Gene diversity H T M FEMSATL FEMSATL FEMSATL FEMSATL intra population diversity --- high --genetic differentiation between strands-- low. -- significant heterozygote deficiency Gene diversity France : Bulgaria

16 Allele number in chloroplast microsatellite regions (haplotypes) in Fraxinus excelsior from Ireland & Europe. Ireland Europe Code SSr-type Size range N H T N H T CPFRAX2 T 9-10 AC 2 T CPFRAX5 T 4 ACTATCT CPFRAX6 T 4 ACTATCT 9-11 & GCGC 3-4 A Multilocus Haplotype N = number of alleles / haplotypes detected H T = is the gene diversity (Nei, 1973). Most variation within provenances ( 79-63%) 14 Irish provenances; Europe from B, Cz. Rep., Dk,UK (S), UK, F, D, L., Poland, Spain.

17 Distribution map of most common haplotypes of Irish ash from provenance trial with new cpdna markers

18 Analysis of provenance relationships Neighbour joining tree of European ash from AFLP data

19 Distribution map of Common haplotypes in European ash

20 Propagation technology Sources of material Selected trees Micropropagation Somatic embryogenesis Cryopreservation Cutting propagation

21 Micropropagation Culture initiation : 2003 Buds from 55 selected clones 18 clones viable (@ 4 subcultures) Medium: (mg/l) M9 = MS : BA 5.5; TDZ 0.55, IBA 0.2 QRC = WPM*with 3.0 g/l charcoal. Rooting spontaneously or induced with auxin

22 Culture establishment is difficult multiplication rate clone 14/63 14/68 14/66 Influence of TDZ on the initiation of shoo cultures WPM with: 14/63: 4mg/l BAP mg/l IBA, 14/68: 4mg/l BAP mg/l IBA + 0.2mg/l TDZ, 14/66: 0.15mg/l IBA + 1mg/l TDZ 4 3,5 Propagation rate 3 2,5 2 1,5 1 0,5 JK49 T70 F Subcultures Micropropagation over time

23 Somatic embryogenesis ( M. Capuana ) MS medium containing 2,4-D (0.1 mg l-1 ) & BA (1.0 mg l-1 ) % germination % conversion

24 Cryopreservation of clones of Fraxinus excelsior clone no. of cryopreserved shoot tips regeneratio n rate [%] juvenile 1- week ( 1-2 mm) weeks adult weeks (A. Meier Dinkel)

25 Cutting propagation ( G. Douglas ) Types of cuttings Rooting cuttings of Ash cuttings from seedlings and micropropagated plants: Apical = Apex + 2 nodes Subapical= 2 nodes

26 Rooting in cuttings from micropropagated ash 2 wks. Rooting: warm bench with plastic 3 months

27 Hedges of stock plants Cuttings Cuttings collected from the same hedge April ( dormant ) May June July

28 Rooting in cuttings from micropropagated hedged plants in May June & July May June July Rooting % F Clone

29 Hedges from micropropagation. Rooted cuttings (85-100%) Four crops of cuttings per year 1-2 yr growth Field planting 400 rooted / m 2 : 4 cutting crops / yr = 1600 rooted / yr / m 2 glasshouse of 200 m 2 = 320, 000 rooted ash plants / yr.

30 Utilisation strategies (A. Ottitsch, J. Frawley) Improved broadleaves Field User Factors of influence?? Planting broadleaves: Farmers survey Focus group discussions

31 Flow of information & materials from research institutes to forestry entrepreneurs model

32 Afforestation environment in Ireland ha / yr. Broadleaf %-- target 30% Species Ash Oak Alder Sycamore Beech Other % Grants and Premia Species Oak Beech Other Total / ha grant Euro Euro p.a. /ha Premium (20 yrs)

33 Farmer survey : Factors which Influence the planting of broadleaf species Main influence Broadleaf owners Broadleaf + conifers owners % % No * All % Higher subsidies Aesthetic/amenity Land suitability Environmental effects Other Total

34 Landholders survey views: planting improved material ( when available ). View on advantages % Advantage definitely Yes, probably No, advantage probably No, definitely Not sure All

35 Education level of landowners by plantation type. Highest education level obtained Broadleaf owner Broadleaf & conifer owner Conifer owner None All % % % % % Primary only Secondary level Third level All

36 Revised model -- Strong role for opinion leaders

37 Thanks to the RAP team & Thank you for your attentio

38