TECHNOLOGY IMPLEMENTATION PLAN

1. Main results of the project

Table 1 summarizes the original aims outlined in the technical annexe as well as the delivered results obtained by the end of the With only one exception, deliverables were provided for all the outputs that were set out at the start of the project. Furthermore, deliverables reached levels well beyond the original expectations. A major achievement of the project was the construction of the synthetic maps at a continental scale. The original intention to monitor cpDNA polymorphism in 1187 populations (see technical annexe), was far exceeded and  2673 populations, representing a total number of 12714 trees was eventually analysed. The greater efforts resulted in an increase in the density of populations per country as well as  an increase of the number of countries sampled. At the beginning of the project 12 countries were participants, whereas the final inventory is based on a systematic sampling in 22 countries. Many partners associated the financial support of the current project to other resources coming mainly from their own country. In some cases they found support for bilateral cooperation with eastern countries, and contributed to the project by extending the sampling of populations in those countries. An additional partner from Hungary joined the project during the last year. Finally other countries not yet covered by the inventory have started with their own means the monitoring of cpDNA polymorphism

Furthermore, additional issues were addressed during the course of the project that led to important results. Surprisingly, it became obvious that the oak species identification  was based on different criteria in different countries. There was clearly a need to  develop a standardized protocol for assessing leaf morphological traits that could be used  to assign species to trees on a continental scale. This protocol resulted in a simple procedure to identify Q. petraea and Q. robur based on a few morphological assessments and that has been tested on 3025 trees from nine different stands growing on contrasting ecogeographical sites. 

Table 1. Results expected, and deliverables .

2. Benefits of the project

            The main aim of the project was to synthesize the information on oak genetic variation on a European scale by combining traditional methods (provenance tests) with new molecular techniques (cpDNA and microsatellite analysis). As shown on Table 1, these results were achieved beyond expectations. There are at least three types of benefits that can be gained from these results.

2.1. Scientific benefits

            The systematic fine scale mapping of cpDNA polymorphism represents an unprecedented achievement in population genetics of plant species. This project also represents the first attempt to associate genetic and palinological information for reconstructing the postglacial history of a colonising species. The benefits from this association to the scientific community of foresters, ecologists, paleobotanists and geneticists are numerous and are not yet fully identified. We will mostly concentrate on those that may have further implications in other disciplines and which have  initiated new EU supported  projects.

Genomic organisation, evolution of chloroplast DNA polymorphism, and population genetics of organellar genomes

            The chloroplast genome was generally thought to be extremely conserved  but the  systematic analysis of cpDNA polymorphism in 6 white oak species has permitted the identification of genomic regions which exhibit high levels of polymorphism . This information can be further used for designing molecular screening techniques in other species. 

Geographic distribution of diversity and conservation issues

The fine scale map affords basic data on where genetic diversity is concentrated, or buildt up. This is solid, scientific information which can be used by forestry or environmental agencies in connection with adaptive traits as a basis on which to develop their conservation policy. Clearly the southern European regions (Iberian and Italian peninsula, Balkan regions) were confirmed as hot spots of diversity. There were, in addition other European zones, where different maternal lineages merged  which also exhibited high levels of diversity.

Refugial zones, recolonisation routes, and future oak migrations

            The geographic distribution of the cpDNA polymorphism together with maps of fossil pollen deposits have allowed the post glacial recolonisation routes to be traced retrospectively. Because oaks are carrier species in European forest ecosystem, the identification of their postglacial recolonization routes may have further implication on the distribution and diversity of  other plant or animal species. Furthermore the recolonisation dynamics that were identified in this project can be considered as indicators of future migration movements of oaks as a result of climatic changes. Certainly the large data set that was accumulated in the course on this project could be used to validate simulations of migration dynamics under various climatic changes.

Establishment of ISPs and gene flow investigations

Nine ISPs (Intensive studied plots) were established during the course of this project. An ISP is a population in which all trees are fingerprinted and mapped. This represents an outstanding experimental investments for analyzing pollen or seed gene flow at a later date.

2.2. Technical benefits

            The synthetic maps that have been constructed in this project are based on populations of native origin. Although the autochthonous origin of populations could not be guaranteed when the sampling and collection of material was done, the distribution of the cpDNA diversity clearly shows strong geographic patterns. It is therefore likely that the maps that were constructed depict the natural migration routes, rather than human interferences. Hence the maps can be considered as reference maps to which any population of « unknown » origin can be compared. These results open broad perspectives for traceability and identification applications.

Retrospective identification of historical seed transfer in oaks; Genetic test of autochthony

Historically oaks have been transferred over long distances in Europe from eastern and southern countries to western countries and this is still going on today on a minor scale. In most countries, the present oak forest is therefore a mixture of predominantly autochthonous material and allochthonous material. In most cases, the existing archives do not permit the historical origin of the seed source to be identified. The synthetic map will allow any population of « unknown origin » to be assigned to an alien or a local source. This type of information will be highly valuable for oak decline surveys and ecological studies, since sensitivity to dieback or diseases may result from a non native origin of the stand. For the first time, it is possible to differentiate between regional autochthonous and non indigenous origins.

Traceability in the seed-plant chain

European regulation on the use of reproductive material recommend that collections for plantations should be based on stock which is of local origin. Long distance seed transfer may lead to adaptation problems of the introduced material. The verification of the geographic origin of a seed lot (or a seedling bench in a nursery) is based today on official documents. The synthetic map of cpDNA haplotypes can be considered as a tool to check the origin. In most cases, the strict identification of the geographic origin is not possible, but the comparisons of the cpDNA fingerprint of the seed lot to the reference map will certainly allow  the exclusion of certain geographic regions as putative origins.

Traceability in the oak wood chain

            The wine barrel industry in France has expressed strong interest in using genetic fingerprints for systematic screening of oak wood based on the geographic origin. Furthermore, it was recently shown that there was still enough chloroplast DNA in dry wood to allow DNA amplification and fingerprinting. Again the cpDNA maps could be used by the wine barrel industry for verifying the geographic origin of the wood used for barrels.

Routine species identification based on morphological characters

            A simple and operational key for rapid identification of species was constructed during the course of this project. It is based on a few leaf morphological characters and is applicable throughout Europe. This should prove useful to field based foresters.

2.3. Economic benefits

            The project also aimed  to rationalize the use of oak genetic resources for their utilization as source material for plantation. Due to the irregularity of flowering in oaks, natural regeneration is usually supplemented by planting. The choice of the genetically adapted seed source is therefore of crucial importance. The comparative analysis of  all provenance tests of Q. petraea established in western European countries has indicated strong latitudinal clinal patterns for bud burst which is an important adaptive trait.  Northern provenances flush later than southern ones at any given site. These results provide a sound, scientific basis for  the statement that seed transfer from South to North includes more risks.. The comparative analysis across sites also indicated low GE interactions, therefore reinforcing the general conclusion on a European scale.

3. Dissemination of the results

3.1. Construction of a web site

A web site devoted to the project has been constructed, that comprises all the results and deliverables that were obtained: http://www.pierroton.inra.fr/Fairoak/

The Website was so far protected with a password, and will be open after publication of the main results of the project in scientific journals. Besides the general description of the project, the technical annexe and the list of participants, other major items available on the website are described in Table 2

Table 2 Outline of the content of the Website

http://www.pierroton.inra.fr/Fairoak

3.2. Publications

3.2.1 Special Issue of Forest Ecology and Management

            A special issue of Forest Ecology and Management entirely devoted to the results of the project is currently in preparation. It comprises 13 contributions and is subdivided in two sections: general and regional papers.

Table 3. Content of the Special issue of Forest Ecology and Management

General papers

Regional papers

3.2.2. Other publications

Cottrell, J.E., Tabbener, H.E., Munro, R.C., Gillies, A.C.M., 2000 Forrest,G.I., Deans, J.D. & Lowe, A.J. (in preparation) Chloroplast DNA variationin British oak populations. Forest Research Annual Report

Csaikl U., König, A. 1999. Genetische Typen der Eiche in Bayern, Tirol und Oberösterreich. BFH Nachrichten 37. Jahrgang, Heft 4 (Quarterly report, 1 p.).

Dam, B.C. van, en S.M.G. de Vries, 1998. In de voetsporen van de eik, postglaciale herkolonisatie-routes. In: De Levende Natuur, 99 (1), 38-41.

Herrán, A., Espinel, S., Goicoechea, P.G., 1999. Utilización del polimorfismo del ADN de cloroplastos para definir regiones de procedencia materna en los robles blancos de la Península Ibérica. Invest. Agr.: Sist. Recur. For. Vol  8(1), 139-150

Fineschi S., B. Demesure, R. Petit 1995. Genetic differentiation in European oak species (section Robur) inferred from cpDNA polymorphism. Giornale Botanico Italiano 129 (1): 44.

Fineschi S., A. Balijja, A. Diarra, G. Scuderi, D. Taurchini, 1998. Chloroplast DNA polymorphism in oak populations (Quercus, section Robur) from Switzerland (Informatore Botanico Italiano 30: 80-82)

Fineschi S., D. Taurchini, G. Scuderi, 1998. Polimorfismo del DNA cloroplastico in popolazioni di Quercus pubescens Willd. dalla Sicilia. Monti e Boschi, 49: 17-20)

König, A. 1996: Untersuchungen zur genetischen Variation bei Eiche. BFH Nachrichten 34. Jahrgang, Heft 2 (Quarterly report, 1 p.).

König, A.; Groppe K.; Ziegenhagen  B. 1998. First results of chloroplast-DNA investigations in German populations of Quercus petraea and Q. robur. In: Diversity and adaptation in oak species. Proc. of the second meeting of WP 2.08.05, Genetics of Quercus, of the IUFRO. University Park (State College), Pennsylvania, U.S.A., Oct 12-17, 1997. Ed.. K. C. Steiner. The Pennsylvania State University, University Park, PA, 1998, p. 102-108.

Kranenborg, K.G. en S.M.G. de Vries, 1999. Provenance research Quercus robus in De Rips. Wageningen: Instituut voor Bos- en Natuuronderzoek. IBN-researchreport 99/4. ISSN 0928-6896.

Olalde, M., Herrán, A., López de Heredia, U., Espinel, S., Goicoechea, P.G.
Molecular diversity of white oaks from the Iberian Peninsula. Communication
to the conference BIOFOR99, held in Vitoria-Gasteiz from 22 to 25 September
1999

Stephan, B. R. 1999 Progress report on the conservation of genetic resources of beech and oak in Germany in the period 1998 -1999. EUFORGEN-Meeting at Birmensdorf 3.to 6.6.1999.

Ziegenhagen B.; Degen B.; König A. 1999. Analyse der räumlichen genetischen Differenzierung von Eichenbeständen in Nordrhein-Westfalen anhand von Chloroplasten-Genmarkern (internal report for the Research Institute of Nordrhine-Westfalia - LÖBF, 11pp.+ annex)

3.3. Conferences

3.3.1. Joint meeting with EUFORGEN

            The final meeting of the project was held in Birmensdorf (May 31st to June 3^rd) and was followed at the same place by a joint meeting with the EUFORGEN network “Social hardwoods” (June 3^rd-June 6^th). This network comprises national representatives from 24 European countries (Austria, Belgium, Bulgaria, Croatia, Czech Republic, Finland, France, Germany, Hungary, Italy, Ireland, Moldova, Lithuania, Luxembourg, the Netherlands, Norway, Portugal, Slovakia Slovenia, Spain, Sweden, Switzerland, Ukraine and the United Kingdom). The EUFORGEN network “Social hardwoods” is a collaborative programme among European countries which aims to promote the effective conservation and the sustainable utilization of oak and beech  resources. Partners to the Fair project shared their results with the EUFORGEN members and discussed their implications in conservation strategies. Extension of the geographic mapping project eastwards was suggested through a specific R&D project to be submitted to the INCO-COPERNICUS programme or to other agencies (IPGRI).

3.3.2. Other conferences

Bordacs, S.2000 Assessing cpDNA diversity in Hungarian oak populations and its sylvicultural aspects.  To be presented  at the Oak2000 Meeting, Zagreb (May 20^th-25^th)

Csaikl U., König, A. 1999. CpDNA-haplotype mapping of white oak populations north of the Alps and in the Danube Valley. Freising. IUFRO-Conference (Division 2 and 7) «Genetic response of forest systems to changing environmental conditions – analysis and management», in Freising from 12. to 17.9.1999 (*oral presentation14.9.1999 ).

Csaikl  U.; König A. 1999.  CpDNA-haplotype mapping of white oak populations north of the Alps and in the Danube Valley. In: Book of Abstracts of the IUFRO Conference «Genetic Response of Forest Systems to Changing Environmental Conditions - Analysis and Management -». University of Munich, Freising, Germany, September 12-17, 1999, p. 40.

Csaikl  U.; König A. 2000. Genetic types in white oak populations north of the Alps and in the Danube valley. Proceedings of the IUFRO-Conference (Division 2 and 7) «Genetic response of forest systems to changing environmental conditions – analysis and management», in Freising from 12. to 17.9.1999, 10pp. (submitted for printing).

Csaikl U. König, A. 2000. The distribution of genetic types in white oak populations in the Danube region of Germany and Austria. Submitted to the IUFRO-Conference of P1.06.00 and S2.08.05 "Oak 2000", Zagreb, Croatia, May 20-25, 2000, 2pp.

Fineschi S., B. Demesure, R. Petit 1995. Genetic differentiation in European oak species (section Robur) inferred from cpDNA polymorphism.   90° Congresso Società Botanica Italiana, Palermo.

Fineschi S., A. Diarra, A. Balijja, G. Scuderi, D. Taurchini, 1997. Polimorfismo del DNA cloroplastico in popolazioni di querce (sezione Robur) in Sicilia: una risorsa genetica da salvaguardare.   I Congresso della Società Italiana di Selvicoltura ed Ecologia Forestale, Padova.

Finkledey, R 2000 Genetische Variation in Eichenbeständen der Schweiz "24. Int. Tagung derArbeitsgemeinschaft für Forstgenetik und Forstpflanzenzüchtung",14.3.-16.3.2000 in Pirna, Germany,

König, A.*; Groppe, K.; Ziegenhagen, B.: First Results of Chloroplast-DNA Investigations in German Populations of Quercus petraea and Q. robur. Meeting on Diversity and Adaptation in Oak Species, in State College, PA/ USA from 12. to 17.10.1997 (*oral presentation: 13.10.1997).

Kremer^, A., R.J. Petit, U. Csaikl, J. L.Beaulieu, S. Bordacs, S. Brewer, K. Burg, R.Cheddadi, K. Burg, E. Coart, J. Cottrell, N. Cundall, D. Deans, S. Fineschi, R. Finkeldey, A. Gillies, P. Goicoechea, J. Gracan, P. Grossoni, C. L. Hallé, J. S. Jensen, J. Kleinschmit, A. König, A. Lowe, S. F. Madsen, G. Matyas ,R. Munro, M. Olalde, I.Oledska, M. H. Pemonge, F. Popescu, M. Reille , D. Slade, S. Steinhoff, H. E. Tabbener, B. Van Dam, S.M.G. de Vries, B. Ziegenhagen 2000

Range wide distribution of the chloroplast DNA and pollen deposits in European  white oaks: inferences about colonisation routes, impact of chloroplastic divergence on phenotypic differentiation. Submitted to the IUFRO-Conference of P1.06.00 and S2.08.05 "Oak 2000", Zagreb, Croatia, May 20-25, 2000, 5pp.

Vries, S.M.G. en B.C. van Dam, 1998. Selection Programma of Oak in The Netherlands. In: K.C. Steiner (ed.). Diversity and Adaptation in Oak Species. Proceedings of the second meeting of working party 2.08.05: Genetics of Quercus, IUFRO. October 12-17 1997. University Park, State College. p.201-208.

4. Exploitation of results

            As outlined in the previous paragraphs, the geographic maps of cpDNA haplotypes (results R1) across Europe can be used as reference map for traceability applications. Commercial applications can therefore be expected in the seed (for seed trade) or wood chain (wine barrel industry for example).

            Results that can lead to commercial exploitation are R1 (Table 1). All partners have contributed to R1 except P12. Because exploitation may not be immediate due to additional R&D activities, intentions to exploit were usually prospective. At this point, three partners (P1, P5, and P11) did not exclude the possibility to exploit the results in the future, Hence results R1 fall under the category A (“Those results that are of commercial interest to the consortium partners who will therefore exploit these results themselves or have them exploited”). P5 and P11 intend to exploit on their own, and P1 intends to exploit the results through a third party. An SME has shown interest in using genetic fingerprints for the wine barrel industry, and will perform additional R&D research on the subject. All partners agreed that the exploitation should be done on a royalty free conditions. Consequently, the partners intending to exploit through a third party, will not request royalties to the third party. Hence the other partners, not intending to exploit the results, will not request royalties to the partners exploiting. The material and information on exploitable results is stored on the website of the project: http://www.pierroton.inra.fr/Fairoak) as maps and protocols and will be made public after the publication of the special issue of Forest Ecology and Management. There will be no intention to protect the results by either a patent or copyright.