S Todea (Morar), I. M., González-Orenga, S., Boscaiu, M., Plazas, M., Sestras, A. F., Prohens, J., … Sestras, R. E. (2020). Responses to Water Deficit and Salt Stress in Silver Fir (Abies alba Mill.) Seedlings. Forests, 11(4), 395. doi:10.3390/f11040395 Sorensen, F. C. (1999). RELATIONSHIP BETWEEN SELF-FERTILITY, ALLOCATION OF GROWTH, AND INBREEDING DEPRESSION IN THREE CONIFEROUS SPECIES. Evolution, 53(2), 417-425. doi:10.1111/j.1558-5646.1999.tb03777.x

Rajora, O. P., & Mosseler, A. (2001). Euphytica, 118(2), 197-212. doi:10.1023/a:1004150525384

[EN] Assessment of the diversity of reproductive traits and genetic variation is of great relevance to the conservation of genetic resources and management of silver fir (Abies alba) populations. We have evaluated reproductive characteristics associated with female cones and seed morphology, as well as seed germination after subjecting seeds to five storage methods, in nine Romanian populations of A. alba. The genetic diversity of the populations was assessed with 12 polymorphic simple sequence repeat (SSR) markers. We detected significant differences between populations for all reproductive traits and considerable differences in seed germination and storage methods; seed storage in wet sand was the method resulting in the highest germination in all populations. Genomic SSRs (gSSRs) were more informative on average than expressed sequence tag SSRs (EST-SSRs) in the populations studied. The nine populations were genetically diverse, with an average number of alleles (N) per SSR locus between 3.50 and 4.83. The observed heterozygosity (Ho) in the nine populations was always lower than the expected heterozygosity (He), which resulted in values of the inbreeding coefficient (Fis) between 0.261 and 0.709. Genetic distances between populations ranged between 0.077 and 0.410. The cluster analysis based on genetic distances did not group accessions according to their geographical proximity, and despite a positive trend, the correlation between geographic and genetic distances was non-significant. The results of an analysis of molecular variance (AMOVA) revealed that only 9.1% of the total molecular variance is attributable to differences between populations. This low degree of genetic differentiation between populations is confirmed by the intermingling of individuals of different populations in a principal coordinate analysis (PCoA). We found evidence of a positive relationship between He and germination, as well as a negative one between Fis and germination, suggesting that populations with low diversity and high consanguinity may have a reduced fitness and long-term viability. The results are relevant for the conservation and management of local genetic resources and populations, as well as for reforestation programmes of silver fir. The publication was supported by funds from the National Research Development Projects to Finance Excellence (PFE)-37/2018-2020 granted by the Romanian Ministry of Research and Innovation. The first author (I.M.T.M.) is thankful to UPV (Valencia) and the Erasmus+ mobility programme, financed by the European Commission, for her scholarship in UPV. The research was partially supported by the Advanced Horticultural Research Institute of Transylvania (ICHAT), University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca (USAMVCN), and Polytechnic University of Valencia (UPV). Mariola Plazas is grateful to Generalitat Valenciana and Fondo Social Europeo for a post-doctoral grant (APOSTD/2018/014). Todea Morar, IM.; Rensen, S.; Vilanova Navarro, S.; Boscaiu, M.; Holonec, L.; Sestras, AF.; Vicente, O.... (2020). Genetic Relationships and Reproductive Traits of Romanian Populations of Silver Fir (Abies alba): Implications for the Sustainable Management of Local Populations. Sustainability. 12(10):1-15. https://doi.org/10.3390/su12104199 Dering, M., Sękiewicz, K., Boratyńska, K., Litkowiec, M., Iszkuło, G., Romo, A., & Boratyński, A. (2014). Genetic diversity and inter-specific relations of western Mediterranean relic Abies taxa as compared to the Iberian A. alba. Flora - Morphology, Distribution, Functional Ecology of Plants, 209(7), 367-374. doi:10.1016/j.flora.2014.03.011 Tinner, W., Colombaroli, D., Heiri, O., Henne, P. D., Steinacher, M., Untenecker, J., … Valsecchi, V. (2013). The past ecology ofAbies albaprovides new perspectives on future responses of silver fir forests to global warming. Ecological Monographs, 83(4), 419-439. doi:10.1890/12-2231.1 Dobrowolska, D., Bončina, A., & Klumpp, R. (2017). Ecology and silviculture of silver fir (Abies alba Mill.): a review. Journal of Forest Research, 22(6), 326-335. doi:10.1080/13416979.2017.1386021 TODEA (MORAR), I. M., GONZÁLEZ-ORENGA, S., PLAZAS, M., SESTRAS, A. F., PROHENS, J., VICENTE, O., … BOSCAIU, M. (2019). Screening for Salt and Water Stress Tolerance in Fir (Abies alba) Populations. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 47(4), 1063-1072. doi:10.15835/nbha47411348 Schutz, J.-P. (2002). Silvicultural tools to develop irregular and diverse forest structures. Forestry, 75(4), 329-337. doi:10.1093/forestry/75.4.329 PLESA, I., DAN, C., TRUȚA, A., HOLONEC, L., SESTRAS, A. F., BOSCAIU, M., & SESTRAS, R. E. (2017). Spruce Trees Growth and Forest Landscape Depending on Microstational Factors and Ecological Conditions. Notulae Scientia Biologicae, 9(4), 582-588. doi:10.15835/nsb9410217 SESTRAS, P., BONDREA, M. V., CETEAN, H., SĂLĂGEAN, T., BILAŞCO, Ş., NAȘ, S., … CÎMPEANU, S. M. (2018). Ameliorative, Ecological and Landscape Roles of Făget Forest, Cluj-Napoca, Romania, and Possibilities of Avoiding Risks Based on GIS Landslide Susceptibility Map. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 46(1), 292-300. doi:10.15835/nbha46110866 Bončina, A., Simončič, T., & Rosset, C. (2019). Assessment of the concept of forest functions in Central European forestry. Environmental Science & Policy, 99, 123-135. doi:10.1016/j.envsci.2019.05.009 Skrzyszewska, K., & Chłanda, J. (2009). A study on the variation of morphological characteristics of silver fir (Abies alba Mill.) seeds and their internal structure determined by X-ray radiography in the Beskid Sądecki and Beskid Niski mountain ranges of the Carpathians (southern Poland). Journal of Forest Science, 55(No. 9), 403-414. doi:10.17221/48/2008-jfs Gradečki-Poštenjak, M. (2016). The influence of crown defoliation on the variability of some physiological and morphological properties of silver fir (Abies alba) seeds in the seed zone of Dinaric beech-fir forests in Croatia. Periodicum Biologorum, 117(4), 479-492. doi:10.18054/pb.2015.117.4.3428 Gömöry, D., Paule, L., Krajmerová, D., Romšáková, I., & Longauer, R. (2012). Admixture of genetic lineages of different glacial origin: a case study of Abies alba Mill. in the Carpathians. Plant Systematics and Evolution, 298(4), 703-712. doi:10.1007/s00606-011-0580-6 Postolache, D., Leonarduzzi, C., Piotti, A., Spanu, I., Roig, A., Fady, B., … Vendramin, G. G. (2013). Transcriptome versus Genomic Microsatellite Markers: Highly Informative Multiplexes for Genotyping Abies alba Mill. and Congeneric Species. Plant Molecular Biology Reporter, 32(3), 750-760. doi:10.1007/s11105-013-0688-7 De Luca, D., Menale, B., Caputo, P., & Cennamo, P. (2016). Population genetics analysis in a relic population of silver fir (Abies alba Mill.) in southern Italy: A comparison with microsatellites and reference data. Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology, 151(4), 567-573. doi:10.1080/11263504.2016.1186124 Belletti, P., Ferrazzini, D., Ducci, F., De Rogatis, A., & Mucciarelli, M. (2017). Genetic diversity of Italian populationsof Abies alba. Dendrobiology, 77, 147-159. doi:10.12657/denbio.077.012 Popovic, V., Lucic, A., Rakonjac, L., Milovanovic, J., Mladenovic-Drinic, S., & Ristic, D. (2019). Application of SSR markers for assessment of genetic differentiation of silver fir (Abies alba Mill.) originating from Javor mountain. ABI Genetika, 51(3), 1103-1112. doi:10.2298/gensr1903103p Popovic, V., Lucic, A., Rakonjac, L., Cvjetkovic, B., Mladenovic-Drinic, S., & Ristic, D. (2017). Assessment of genetic diversity of silver fir (Abies alba Mill.) in Serbia using SSR markers. ABI Genetika, 49(3), 979-988. doi:10.2298/gensr1703979p Donohue, K., Rubio de Casas, R., Burghardt, L., Kovach, K., & Willis, C. G. (2010). Germination, Postgermination Adaptation, and Species Ecological Ranges. Annual Review of Ecology, Evolution, and Systematics, 41(1), 293-319. doi:10.1146/annurev-ecolsys-102209-144715 Slate, J., David, P., Dodds, K. G., Veenvliet, B. A., Glass, B. C., Broad, T. E., & McEwan, J. C. (2004). Understanding the relationship between the inbreeding coefficient and multilocus heterozygosity: theoretical expectations and empirical data. Heredity, 93(3), 255-265. doi:10.1038/sj.hdy.6800485 Caudullo, G., Welk, E., & San-Miguel-Ayanz, J. (2017). Chorological maps for the main European woody species. Data in Brief, 12, 662-666. doi:10.1016/j.dib.2017.05.007 HANSEN, O. K., VENDRAMIN, G. G., SEBASTIANI, F., & EDWARDS, K. J. (2005). Development of microsatellite markers in Abies nordmanniana (Stev.) Spach and cross-species amplification in the Abies genus. Molecular Ecology Notes, 5(4), 784-787. doi:10.1111/j.1471-8286.2005.01062.x CREMER, E., LIEPELT, S., SEBASTIANI, F., BUONAMICI, A., MICHALCZYK, I. M., ZIEGENHAGEN, B., & VENDRAMIN, G. G. (2006). Identification and characterization of nuclear microsatellite loci in Abies alba Mill. Molecular Ecology Notes, 6(2), 374-376. doi:10.1111/j.1471-8286.2005.01238.x Peakall, R., & Smouse, P. E. (2012). GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research--an update. Bioinformatics, 28(19), 2537-2539. doi:10.1093/bioinformatics/bts460 Jombart, T., & Ahmed, I. (2011). adegenet 1.3-1: new tools for the analysis of genome-wide SNP data. Bioinformatics, 27(21), 3070-3071. doi:10.1093/bioinformatics/btr521 Kamvar, Z. N., Tabima, J. F., & Grünwald, N. J. (2014). Poppr: an R package for genetic analysis of populations with clonal, partially clonal, and/or sexual reproduction. PeerJ, 2, e281. doi:10.7717/peerj.281 Schliep, K., Potts, A. J., Morrison, D. A., & Grimm, G. W. (2017). Intertwining phylogenetic trees and networks. Methods in Ecology and Evolution, 8(10), 1212-1220. doi:10.1111/2041-210x.12760 Nei, M. (1972). Genetic Distance between Populations. The American Naturalist, 106(949), 283-292. doi:10.1086/282771 Excoffier, L., Smouse, P. E., & Quattro, J. M. (1992). Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics, 131(2), 479-491. doi:10.1093/genetics/131.2.479 Verdú, M., & Traveset, A. (2005). EARLY EMERGENCE ENHANCES PLANT FITNESS: A PHYLOGENETICALLY CONTROLLED META-ANALYSIS. Ecology, 86(6), 1385-1394. doi:10.1890/04-1647 Cremer, E., Ziegenhagen, B., Schulerowitz, K., Mengel, C., Donges, K., Bialozyt, R., … Liepelt, S. (2012). Local seed dispersal in European silver fir (Abies alba Mill.): lessons learned from a seed trap experiment. Trees, 26(3), 987-996. doi:10.1007/s00468-012-0676-9 Carles, S., Lamhamedi, M. S., Beaulieu, J., Stowe, D. C., Colas, F., & Margolis, H. A. (2009). Genetic Variation in Seed Size and Germination Patterns and their Effect on White Spruce Seedling Characteristics. Silvae Genetica, 58(1-6), 152-161. doi:10.1515/sg-2009-0020 Bussotti, F., Pollastrini, M., Holland, V., & Brüggemann, W. (2015). Functional traits and adaptive capacity of European forests to climate change. Environmental and Experimental Botany, 111, 91-113. doi:10.1016/j.envexpbot.2014.11.006 Teodosiu, M., Mihai, G., Fussi, B., & Ciocîrlan, E. (2019). Genetic diversity and structure of Silver fir (Abies alba Mill.) at the south-eastern limit of its distribution range. Annals of Forest Research, 62(2), 139. doi:10.15287/afr.2019.1436 Pandey, M. K., Gautami, B., Jayakumar, T., Sriswathi, M., Upadhyaya, H. D., Gowda, M. V. C., … Varshney, R. K. (2011). Highly informative genic and genomic SSR markers to facilitate molecular breeding in cultivated groundnut (Arachis hypogaea). Plant Breeding, 131(1), 139-147. doi:10.1111/j.1439-0523.2011.01911.x Martin, M. A., Mattioni, C., Cherubini, M., Taurchini, D., & Villani, F. (2010). Genetic diversity in European chestnut populations by means of genomic and genic microsatellite markers. Tree Genetics & Genomes, 6(5), 735-744. doi:10.1007/s11295-010-0287-9 Muñoz-Falcón, J. E., Vilanova, S., Plazas, M., & Prohens, J. (2011). Diversity, relationships, and genetic fingerprinting of the Listada de Gandía eggplant landrace using genomic SSRs and EST-SSRs. Scientia Horticulturae, 129(2), 238-246. doi:10.1016/j.scienta.2011.03.034 Nandha, P. S., & Singh, J. (2013). Comparative assessment of genetic diversity between wild and cultivated barley using gSSR and EST-SSR markers. Plant Breeding, 133(1), 28-35. doi:10.1111/pbr.12118 VILCAN (TRUTA), A., MIHALTE (CRISAN), L., SESTRAS, A. F., HOLONEC, L., & SESTRAS, R. E. (2017). Genetic variation and potential genetic resources of several Romanian larch populations. TURKISH JOURNAL OF AGRICULTURE AND FORESTRY, 41, 82-91. doi:10.3906/tar-1610-57 GRAMAZIO, P., PLESA, I. M., TRUTA, A. M., SESTRAS, A. F., VILANOVA, S., PLAZAS, M., … SESTRAS, R. E. (2018). Highly informative SSR genotyping reveals large genetic diversity and limited differentiation in European larch (Larixdecidua) populations from Romania. TURKISH JOURNAL OF AGRICULTURE AND FORESTRY, 42(3), 165-175. doi:10.3906/tar-1801-41

Kaliniewicz, Z., Markowski, P., Anders, A., Jadwisieńczak, K., Żuk, Z., & Krzysiak, Z. (2019). Physical Properties of Seeds of Eleven Fir Species. Forests, 10(2), 142. doi:10.3390/f10020142

Kaliniewicz, Z., Markowski, P., Anders, A., Jadwisieńczak, K., Żuk, Z., & Krzysiak, Z. (2019). Physical Properties of Seeds of Eleven FIR Species. Forests, 10(2), 142. doi:10.3390/f10020142

Genetic Relationships and Reproductive Traits of Romanian Populations of Silver Fir (Abies alba): Implications for the Sustainable Management of Local Populations

Abstract