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Activities on forest gene concervation and tree breeding in Lithuania

...Conservation of genetic diversity is the key element in conservation of biodiversity ! .......

Present status of gene conservation and tree breeding

Genetic diversity, which characterises the inter- and intra-population diversity within species, aside the ecosystem diversity and species diversity is an important level of bio-diversity. Therfore the conservation of genetic diversity is considered as the key element in conservation of biodiversity and sustainable forestry. However, in Lithuania, the Law of Protected Territories, the Law of Forest, and the Law of Wild Plants, etc., for a long period have not recognised the importance of gene conservation. There were no specialised laws for protection and conservation of forest genetic resources. Botanical reserves and other types of reserves were aimed at preserving endangered species and their biotopes, but are not concerned with conserving genetic diversity as such. In Lithuania, units for conservation of forest genetic resources were not included into register of the territories protected at state level. Only in 2002, the new law on Plant national genetic resources and updated Forest law and law on Protected areas recognized the importance of conservation forest genetic resources.

Conservation of forest genetic resources is co-ordinated and financed by the Department of Forests under the Ministry of Environment of the Lithuanian Republic. The Conception Program for Forest Regeneration that was approved in 1994 includes the prospective development of a basis for conservation of genetic diversity. The Convention for Protection of Bio-diversity signed in June 1992 in Rio de Janeiro was ratified in 1995. The State Program on Lithuanian Forestry and Timber Industry Development (approved in 1996 for period up to 2003) and the Strategy and Plan for Action for Protection of Bio-diversity in Lithuanian Republic (approved in 1997) consider some aspects of forest gene conservation. However, there were no special national program concerned with the development of applied gene conservation and breeding. Just recently (in 2003) the Program for conservation of forest genetic resources was adopted for the 10 year period (2004 to 2014).

During the past 40 years a wide network of units for conventional gene conservation and tree breeding has been created in Lithuania. At present (2003 01 01), the network of forest gene conservation in situ and ex situ, and tree breeding units consists of:

194 genetic reserves (4623 ha),
126 seed reserves (1062.1 ha),
1080 plus trees,
7 archives of clones (714 clones, 38.3 ha),
130 progeny test plantations (203 ha)
94 seed orchards (657.3 ha).

Genetic reserves are designated to preserve genetic diversity of a species over environmentally heterogeneous sites on comparatively small areas (less than 30 ha) by restriction of silvicultural activities. Strict genetic reserves that were established two decade ago to preserve sustainable development of resources in communities of species in environment that is as natural as possible over a large forest area (more than 200 ha) have fail to achieve their destiny therefore recently they were transformed into genetic reserves. The main efforts were placed on Scots pine and Norway spruce. Past work in establishing conventional gene conservation network that safeguarded important populations from cutting should be considered as the first step in creating a functional gene conservation system. Now we need to proceed further in order to secure sustainable gene conservation.

		Unfavourable environmental conditions in recent years showed drawbacks in the conventional in-situ conservation system. Over half of the coniferous genetic resources and a portion of the broad-leaved genetic resources were lost due to combined action of droughts, insect outbreaks, and windfalls, or due to natural succession of tree species. In the period 2001 to 2002 huge areas of ash stands were efected by dieback all over the country. It indicates that we have already experiencing negative consequences of global climatic changes. Based on experience and on new understanding of general gene conservation objectives and principles, the shortcomings of the conventional static forest gene conservation system are outlined below.
Photos by Romas Barauskas

1. In the past, the non-interference approach and management restrictions were based on preservationist conservation objectives were not adequate to the current rapidly changing environment. Therefore the:

sustainability of gene conservation populations is low;
regeneration of conservation population is not guaranteed;
existing genetic constitution is ‘frozen’; and
use of regeneration material from the best genetic resources in forestry is very limited.

2. Natural genetic diversity was insufficiently represented from geographical, population, ecological, and generation viewpoints. In the past:

principles, criteria and methods used for selecting and sampling of genetic resources were based mostly on the economic utility of tree breeding objectives;
geographical patterns of tree species variation were neglected in selecting, sampling, and conservation;
geographical eco-climatic patterns were neglected; and
only the oldest generation is represented in gene conservation areas.

3. Legislation, financing, and publicity were insufficient to accomplish gene conservation objectives:

insufficient legislation for gene conservation of forest tree species;
inadequate financing to conserve a large number of gene conservation populations;
insufficient education of foresters on gene conservation; and
insufficient public awareness.

These shortcomings call for the urgent development of a new system for sustainable and secure gene conservation that would be flexible enough to cope with rapid environment changes and expansion of human activities into forestlands.

In 1994, Lithuanian Forest Research Institute in co-operation with other Lithuanian agricultural institutes and universities have started research programe "Plant genetic resources", Lithuania joined to the EUFORGEN program, Nordic gene Bank has started program for support of conservation of genetic resources in Baltic states, in 1998, the long term state research program GENOFUND have been launched. These programs have promoted the re-activating and initiating of new activities regarding forest gene conservation and tree breeding in Lithuania. One of many positive issues of these programs was that they convinced and motivated national authorities to support preparation of new legislation on gene conservation, tree breeding, seed farming, reforestation, etc.

Norway spruce seed reserve in Kurtuvenai

Participation in activities of three EUFORGEN networks has helped to increase the understanding of new approaches in forest gene conservation and to adjust to new concepts in the preparation of new national strategies, programs and recommendations. As outlined below, there has been much recent progress in forest gene conservation and tree breeding during last decade in Lithuania.

In 2003, the Program for conservation of forest genetic resources and tree breeding was adopted for the 10 year period (2004-2014) of activities.
In 2003, provenance regions were delineated and aproved for Alnus glutinosa, Acer platanoides, Tilia cordata, and Populus tremula as well as regulations for use and transfer of reproductive material (Order of Minister of Environment No 54, January 30, 2003).
In the period 1998 to 2003, the state research program GENOFUND "Studies and conservation of plants and animals used in Lithuania" is operating.
In 2002, based on EUFORGEN strategies and guidelines for conservation forest genetic resources in Europe, the project of Program for conservation of genetic resources and breeding of broadleaved tree species was prepared.
In 2002, the Law on National Plant Genetic Resources was adopted.
In 2002, new Biotechnology Laboratory was established in Lithuanian Forest Research Institute.
In 2002, the Department of Genetically Modified Organisms and the Committee for Controll of Management of Genetically Modified Organismswas established under the Ministry of Environment.
In 2002, the Lithuanian Plant Gene Bank was established. It will coordinate activities on conservation of plant genetic resources and will create germampasm collections. The Coordinating Centres of the Lithuanian Plant Gene Bank will be estabished in all institutions dealing with conservation of plant genetic resources.
In 2002, provenance regions were delineated and aproved for Betula pendula, Quercus robur, Picea abies, Pinus sylvestris and Fraxinus excelsior as well as regulations for use and transfer of reproductive material (Order of Minister of Environment No 317, June 14, 2002)..
In 2001, new Law of Forests was adopted with integrated issues on forest gene conservation.
In 2001, new Law of Protected Territories was adopted with integrated issues on forest gene conservation.
In 2000, seed zoning of Pinus sylvestris, Picea abies and Quercus robur was adopted
In 2000, periodic inventory of all forest gene reserves, plus trees, seed stands, and seed orchards was acompished.
In 1999, the preparation of new Law on National Plant Genetic Resources was started..
In 1999, based on EUFORGEN recommendations, the national technical guidelines for dynamic in-situ conservation of Norway spruce and non-rare Noble Hardwoods were prepared and adopted.
In 1999, the Regulations on Forest Genetic Reserves were updated and adopted.
In 1999, the set of PC databases on Lithuanian forest genetic resources were joined and a Joint Database on Forest Genetic Resources was created.
The National Register of Forest Genetic Resources is issued annually based on information in the updated databases.
In 1999, a research project titled “Eco-genetic variation and gene conservation of broad-leaved tree species” (1999-2003) was begun.
In 1998, a State Research Program "Plant genetic resources" (1998-2002) was started.
In 1998, extended Internet Site on Lithuanian Forest genetic Resources was established.
In 1997, the National Gene Bank of Plants, that include the forest genetic resources, was established with technical assistance and financial support from Nordic Gene Bank.
In 1997, for developing of Forest gene Conservation Program the startegies and programs-methods were prepared for dynamic gene conservation of Pinus sylvestris, Betula pendula, Alnus glutinosa, and Fraxinus excelsior.
In 1997, it was started the preparation of National Forest Gene Conservation and Breeding Program based on Multiple Population Breeding System (MPBS) concept.
In 1997, a seed zoning of Pinus sylvestris, Picea abies and Quercus robur was prepared.
In 1997, the Catalogue of Lithuanian plant genetic resources that includes the chapter on forest genetic resources was published in the English language.
In 1997, new Seed Regulations were prepared and adopted. These regulations deal with establishment and usage of forest genetic resources, and with documentation of forest regeneration material in accordance with OECD Scheme for the Certification of Forest Reproductive Material Moving in International Trade.
Some concepts of forest gene conservation were integrated into the Law on Preservation of Bred Varieties and Seed Management (1996), the Law of Preservation of Animals, Plants and Mushroom Species and Communities (1997), and the Law of Natural Flora (1998).
In 1996, the Regulations on Forest Genetic Reserves were prepared and adopted
In 1996, the set of PC databases on Lithuanian forest genetic resources was created.
In 1996, a research project titled “Evaluation of adaptation of quality of Norway spruce populations, gene conservation and use" (1996-2000) funded from the State Buget was begun.
In 1995, it was established the Expert Commission for Lithuanian Forest Genetic Resources and Seed Farming.
In 1995, it was established the Commission of National Genetic Resources.
In 1994, a national collaborative project titled "Genetic Resources of Cultivated Plants" (1994-1997) was started

At present, the main emphasis of forest gene conservation and tree breeding in Lithuania includes:

developing legislation on gene conservation and documentation of regeneration material;
developing the strategies, programs and technical guidelines necessary for dynamic gene conservation;
transforming the conventional national systems of gene conservation and tree breeding into a dynamic multiple population joint breeding and gene conservation system;
continuing research on forest genetic resources;
inventorying and complete documenting of forest genetic resources;
controlling status, management, and use of forest gene conservation and tree breeding units,;
documenting and controlling the origin, breeding value and technical quality of forest reproductive material;
integrating the gene conservation principles into regular, sustainable forestry;
developing freely accessible on INTERNET the information system on forest genetic resources.

Challenges of gene conservation and tree breeding

The prime objective of gene conservation is to ensure the continuous survival, adaptation, and evolution of a species over unlimited number of generations in a continuously changing environment. The objective of breeding of broad-leaved tree species is to improve human utility features, including: (1) quality of stem and wood, (2) growth, (3) resistance and adaptability while conserving a sufficient genetic variation for each successive cycle of long-term selection, taking into consideration future changes in breeding priorities and climate. In order to achieve these objectives, it is necessary to promote the maintenance of a broad genetic variation and to create favourable conditions for rapid adaptation of each species. One general prerequisite for successful evolution is the regeneration of the gene resource population. Thus, active measures should be taken where there is a difficulty in of maintaining the gene resource population over successive generations.

The conventional static forest gene conservation system of the past is being transformed into a dynamic one that is based on the Multiple Population Breeding System (MPBS) concept. The MPBS concept combines secure and sustainable conservation of forest genetic resources, preparation for possible eco-climatic changes, and efficient tree breeding. The corresponding MPBS scheme for conservation of Scots pine in Lithuania has already been developed (Eriksson & Pliūra 1997). Basically, the same parameters would hold for broad-leaved tree species, with some adjustments based on the results of ongoing research. Knowledge of the pattern and degree of the genetic variation of broad-leaved species that are present in a region will serve as a basis for the national forest gene conservation strategies and programs.

According to the MPBS, a gene resource conservation/breeding population should consist of 10-20 small sub-populations, each with an effective population size of 50 genetic entries. At present, it is assumed that at that size of breeding-gene conservation population (750-1100 individuals) the alleles of frequencies down to 0.01 (uncommon + common alleles) will be sampled and that will provide sufficient genetic variation for both long-term sustainable gene conservation and long-term breeding purposes. In creating breeding lines (with the 10 best families in each) alleles of frequencies up to 0.25 are involved and are sufficient to guarantee diversity and sustainability of new stands, and to provide high genetic gain. In order to capture the adaptations that already are present in different eco-climatic conditions, selecting and sampling should sufficiently cover geographical patterns of species genetic variation as well as eco-climatic conditions. The essence of dynamic gene conservation by using the MPBS concept is to promote adaptation by exposing the gene resource population to natural selection and in turn to evolution in a variety of directions (Fig. 2). Therefore, a gene conservation network of both natural in situ sub-populations as well as synthetic ones ex situ ones ought to be established over a broad array of eco-climatic conditions.

Gene conservation populations should be intensively managed to improve the adaptation of each sub-population, to increase genetic differences between them, to guarantee their sustainability during all periods of ontogenesis, to insure continuous regeneration of the population of the target species, and to protect against all types of damage. In order to minimise costs, gene conservation ought to be carried out jointly with tree breeding and genetic studies.

Recently EUFORGEN has recommended the levels (milestones) of gene conservation activities that ought to be followed on a step-by-step or parallel manner. These levels are:

defining eco-geographic zones (“seed zones”);
performing inventories of actual species distribution and conservation status;
promoting afforestation using local material within an eco-geographic zone (unless genetic knowledge suggests otherwise);
implementing in situ conservation measures in managed stands with at least 100 regularly fruit producing trees each;
selecting at least 30 stands throughout the distribution area and in nature reserves (5-10 in Europe) in order to create a European network of gene conservation stands;
designating one gene conservation stand in each eco-geographic zone; and
establishing ex situ conservation and seed production areas.

In order to successfully develop these gene conservation measures and to transform the conventional national system of gene conservation and tree breeding into a dynamic multiple population joined breeding - gene conservation system, we must develop and adopt legal and policy instruments that promote and encourage forest gene conservation in state and private forests. We must:

completely integrate the concepts of dynamic forest gene conservation into existing and new national legislation on forestry, the environment, nature protection, etc. (e.g., the Law on Preservation of the Natural Flora, the Law of Protected Territories; the Law on Conservation of Plant Genetic Resources, and the Law of Forests);
develop and adopt the national strategies for dynamic gene conservation, national programmes and a logical framework for applied conservation and tree breeding for all tree species;
develop and adopt criteria and indicators on sustainable forest management relating to forest genetic resources in order to use them in policy analysis;
develop the recommendations (guidelines) which ensure that forest management and regeneration respect the gene conservation requirements;
create a special national fund for supporting the activities on research and applied forest gene conservation and tree breeding;
develop and adopt legal and economic policy instruments in support of using appropriate forest reproductive material in state and private forests;
establish the designated sub-unit at the Department of Forest Protected Territories under the Ministry of Environment of the Lithuanian Republic in order to monitor and co-ordinate at national level the activities on forest regeneration, gene conservation and tree breeding;
increase the level of knowledge among foresters on basic genetic requirements and gene conservation;
increase the level of general public awareness on forest gene conservation;
seek international collaboration in joint research projects with financial and technical support or assistance by other countries or international organisations and funds.

Constructing a joint gene conservation and tree breeding program

According to occurrence, distribution, population size, social status, stage in ecosystem, and economic importance in Lithuania, tree species could be grouped into 4 groups: (1) common coniferous tree species of high economic importance , (2)common social broad-leaved tree species of moderate economic importance ; (2) for uncommon social broad-leaved tree species; and (3) for rarely occurred asocial broad-leaved tree species of minor economic importance. Some of species characteristics differ from those that species demonstrate in Nordic countries or West Europe.

Four different gene conservation and breeding programs that are based on MPBS and evolutionary approach are under preparation for these groups of broad-leaved tree species:

for the common coniferous tree species that are going to be intensively bred and conserved (Pinus sylvestris, Picea abies) intensively managed 12-16 in-situ (2-3 in each of 6 breeding zones) and 7-10 ex-situ gene conservation-breeding sub-populations are planned;
for the common social broad-leaved tree species that are that are going to be moderately bred and conserved (Alnus glutinosa, Betula pendula, Fraxinus excelsior, Quercus robur), intensively managed 10-12 in-situ (2 in each of 6 breeding zones) and 4-7 ex-situ gene conservation-breeding sub-populations are planned;
for the uncommon social broad-leaved tree species (Quercus petrea, Tilia cordata, Acer platanoides, Fagus sylvatica), that are going to be conserved and bred with low intensity, 6-10 in-situ and 2-3 ex-situ gene conservation sub-populations will be established over breeding zones where the species are more abundant.
for the uncommon rare asocial broad-leaved tree species of minor present economic importance (Ulmus spp, Prunus avium, Carpinus betulus, Malus spp., Pyrus spp., Sorbus aucuparia), 3-6 in-situ and 1-2 ex-situ gene conservation-breeding sub-populations as a progeny test plantations or clonal archives in breeding zones where that species is more common are planned. The additional planting of these rare species could be done in openings within the forest or in forest margins in in-situ gene conservation populations designated for gene conservation of other broad-leaved tree species.

The networks of gene conservation sub-populations are under creation along defined eco-climatic gradients and forest eco-regions (=regions of provenances) in Lithuania .

In-situ gene conservation sub-populations are under establishment on areas that already have the most suitable conventional gene reserves (Fig. 2). In the regions that have no suitable gene and seed reserves some new sub-populations should be selected. The sub-populations should represent: (a) the forests of the main eco-regions (regions of provenance or breeding zones); (b) marginal populations; (c) populations valuable for breeding; (d) endangered populations; (e) populations that have rare or distinctive features; and (f) populations growing under specific ecological conditions.

The networks of ex-situ sub-populations are being established as regular progeny test plantations. In order to safeguard the evolutionary potential of the species and increase the efficiency of tree breeding the population size of ex-situ gene conservation-breeding sub-populations should be of 200-300 genetic entries. The genetic diversity of ex-situ sub-populations could be increased using material from neighbouring eco-regions and countries. The new generations will be created using open pollination or crossing the best 50 individuals selected within the best 50 families.

The combination of methods for mating, testing, selecting and creating production populations in constructing tree breeding-gene conservation ex-situ programs is selected based on species eco-genetic peculiarities, species economic importance and financial possibilities. Due to financial limitations there is no realistic to plan to use a high number of families from plus trees and representatives of natural sub-populations, and some very intensive methods (e.g., artificial crossing, testing of vegetative progeny, vegetative propagation, etc.) in the first cycles of breeding broad-leaved tree species. The principle scheme (with some alternatives) of multiple-population long-term breeding-gene conservation of non-rare broad-leaved tree species in Lithuania is presented in Figure

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