Populus breeding and cultivation studies in Lithuanian Research Centre for Agriculture and Forestry

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1 Populus breeding and cultivation studies in Lithuanian Research Centre for Agriculture and Forestry Jonas Žiauka Lithuanian Research Centre for Agriculture and Forestry Institute of Forestry

2 2 Lithuanian Research Centre for Agriculture and Forestry (LRCAF) since 2010

3 3 Forests in Lithuania: 2.18 million ha (33.5% of the country s territory) Kaunas LRCAF Institute of Forestry Girionys, Kaunas District Vilnius

4 4 Departments of the Institute of Forestry Silviculture department; Ecology department; Forest tree genetics and breeding department (research scientists: V. Baliuckas, A. Pliūra); Forest protection and game management department; Forest resources, economics and policy department; Forest plant biotechnology laboratory (research scientists: S. Kuusienė, J. Žiauka, A. Gradeckas).

5 5 Role of LRCAF Institute of Forestry in hybrid aspen breeding and cultivation

6 6 Native Populus tremula in Lithuanian forests Increase of European aspen (Populus tremula) stands in Lithuania Forest stands area by dominant tree species (2015)

7 7 Institute of Forestry is making genetic research of aspen populations in Lithuania. Three provenance regions for Populus tremula were determined. Also, the best aspen trees for further crosses and progeny field trials are being selected.

8 8 History of hybrid aspen breeding in Lithuania After some test crosses between Populus tremula and two other species of the genus (Populus alba and P. tremuloides), the first hybrid aspen test fields were established in 1968 by Dr. Murkaitė and her team. Second series of crosses was made by Dr. Pliūra in The same Populus species were used (P. tremula, P. tremuloides, P. alba), but the number of trees was extended and more genetic parameters were extracted.

9 9 First hybrid aspen test trials in Lithuania that served as a basis for further breeding experiments (authors: Dr. Murkaitė 1968; Dr. Pliūra ) Year of est. Area, ha Crossings No. of selected trees Populus tremuloides P. tremula P. tremula P. alba, P. tremula P. canescens, P. tremuloides P. tremula P. tremuloides P. tremula, P. tremuloides P. alba, P. tremula P. alba, P. alba P. tremula P. tremuloides P. tremula, P. alba P. tremula P. tremuloides P. tremula, P. tremula P. alba, P. alba P. tremula P. tremuloides P. tremula, P. alba P. tremula In total:

10 10 Hibr drebule Ap tvaras Hybrid aspen (Populus tremuloides P. tremula) test field No. 1 (established in 1983) Hybrid aspen (Populus tremuloides P. tremula and P. alba P. tremula ) test field No. 4 (established in 1968)

11 11 Future plans of LARCF Institute of Forestry in relation to aspen breeding: To select best phenotypes in progeny test plantations of aspen hybrids and to establish collection of best performing parental trees To find the least susceptible trees for Phellinus tremulae, which is known to be the major pathogen that causes stem rot in this species. To propagate selected trees by in vitro culture To establish clonal test trials of best phenotypes To test best trees for short rotation plantations To establish clonal trials of cultivars obtained from crossings among poplars P. trichocarpa, P. nigra, and P. deltoides and to compare their performance to that of aspen hybrids.

12 12 Aspen and its hybrids model species in the work of Forest Plant Biotechnology Laboratory

13 13 Forest Plant Biotechnology Laboratory (Started in 2001 as Laboratory of Molecular Genetics and Biotechnology) Research directions: characterization of forest plant genetic diversity by DNA techniques; research of forest tree embryogenesis and organogenesis in tissue culture; evaluation of growth regulators influence on plant development; development of cloning technologies and bringing them into operation for forest tree multiplication programmes.

14 14 Laboratory s contribution to hybrid aspen breeding and cultivation 1. Molecular (DNA analysis) methods for genetic research Also used for the identity control of clones

15 15 Laboratory s contribution to hybrid aspen breeding and cultivation 2. In vitro propagation of selected trees through microshoot culture

16 16 Mass production of hybrid aspen clones in cooperation with joint-stock company Euromediena ( ) Sterile microshoot culture Acclimatization / rooting 3 weeks In ~6 months 2-4 weeks Clonal plantation Containers in the nursery Green-house 3 months 2-3 weeks

17 17 Laboratory s contribution to hybrid aspen breeding and cultivation 3. Investigation of plant hormone effects in different Populus genotypes and under different culture conditions DPL037 (asp.) DPL038 (asp.) DF1001 (hybr. asp.) Example: study on gibberellin (GA 4/7 ) effects on shoot multiplication

18 18 Laboratory s contribution to hybrid aspen breeding and cultivation 4. Manipulation with artificial environment conditions in order to obtain increased shoot multiplication on hormonefree medium Published research articles: Žiauka J., Kuusienė S., Šilininkas M. (2013) Fast growing aspens in the development of a plant micropropagation system based on plant-produced ethylene action. Biomass and Bioenergy 53: Žiauka J., Kuusienė S. (2014) Multiplication and growth of hybrid poplar (Populus alba P. tremula) shoots on a hormone-free medium. Acta Biologica Hungarica 65:

19 19 Laboratory s contribution to hybrid aspen breeding and cultivation 5. Searching for methods to increase viability and growth of hybrid aspen plants during ex vitro adaptation

20 20 Laboratory s contribution to hybrid aspen breeding and cultivation 6. Production of new aspen hybrids under laboratory conditions, using previously selected hybrid trees 2. Cultivation of isolated ovaries in vitro 1. Fertilization under controlled conditions 3. Primary testing of the offspring in in vitro culture 4. Testing of the offspring in the field

21 21 Results in the already established field trials (2008; combination: (Populus tremuloides P. tremula) (P. alba P. tremula); 500 hybrid seedlings from in vitro culture) showed that newly obtained hybrids have huge phenotypic differences among themselves and provide a possibility to select new trees with particularly good quality. Best 8 (10)-year-old individual: height 12.7 (16.6) m ; stem diameter 20.2 (22.8) cm.

22 Length, mm 22 Different Populus genotypes have different shoot to root ratio during in vitro culture Shoot length Root length Populus alba P. tremula hybrid is different is this aspect both from aspen and hybrid aspen 0 No. 1 (P.tr.) No. 2 (P.tr.) No. 3 (P.tremuloides x P.tr.) No. 4 (P.alba x P.tr.) Experiment with shoot explants of four genotypes on the hormone-free medium

23 Root number per explant 23 Laboratory s contribution to hybrid aspen breeding and cultivation 7. Research of hybrid aspen-associated bacteria and their contribution to plant development 2,8 2,4 2 1,6 Kontrolė Bakterija 1 Bakterija 2 ** * ** ** 1,2 0,8 0,4 0 2 sav. 4 sav. 6 sav. Time after inoculation (in weeks) Paenibacillus sp. increasing aspen root formation

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