Stronger genetic differentiation among within-population genetic groups than among populations in Scots pine provides new insights into within-population genetic structuring.

We investigated the presence of spatial genetic groups within forest tree populations and determined if the genetic divergence among these groups is greater than that between populations using Scots pine (Pinus sylvestris) as a model species. We genotyped 890 adult trees of Scots pine in six natural populations in Lithuania at 11 nuclear microsatellite loci. We used a Bayesian clustering approach to identify the within-population genetic groups within each of the six populations. We calculated the differentiation indexes among the genetic groups within each population and among the six populations by ignoring the genetic groups. The Bayesian clustering revealed 2 to 6 distinct genetic groups of varying size as the most likely genetic structures within populations. The genetic differentiation indexes among the genetic groups within populations were nearly tenfold greater (FST = 0.012-0.070) than those between the populations (FST = 0.003). We conclude on the existence of markedly stronger structuring of genetic variation within populations than between populations of Scots pine in large forest tracts of northern Europe. Such genetic structures serve as a contributing factor to large within population genetic diversity in northern conifers. We assume that within population mating in Scots pine is not completely random but rather is stratified into genetic clusters. Our study provides pioneering novel key insights into structuring of genetic variation within populations. Our findings have implications for examining within-population genetic diversity and genetic structure, conservation, and management of genetic resources.

Can a Forest Tree Species Progeny Trial Serve as an Ex Situ Collection? A Case Study on Alnus glutinosa.

Scientifically informed decisions for the long-term conservation of extant genetic diversity should combine in situ and ex situ conservation methods. The aim of the present study was to assess if a progeny plantation consisting of several open pollinated (OP) families and established for breeding purposes can also serve as an ex situ conservation plantation, using the case study of a Lithuanian progeny trial of Alnus glutinosa, a keystone species of riparian ecosystems that warrants priority conservation actions. We employed 17 nuclear microsatellite (Simple Sequence Repeat) markers and compared the genetic diversity and copy number of the captured alleles of 22 OP progeny families from this plantation, with 10 wild A. glutinosa populations, originating from the two provenance regions of the species in Lithuania. We conclude that the progeny plantation could be used as an ex situ plantation for the A. glutinosa populations from the first provenance region (represented by eight genetic conservation units (GCU)). Based on the present study's results, we can expect that the A. glutinosa progeny plantation harbors enough genetic diversity of wild A. glutinosa populations from the first provenance region. This progeny plantation can serve as a robust ex situ collection containing local alleles present in at least one wild population with at least 0.05 frequency with 25 replications.

Dendrological Secrets of the Pazaislis Monastery in Central Lithuania: DNA Markers and Morphology Reveal Tilia × europaea L. Hybrids of an Impressive Age.

We benefited from the availability of a species-specific DNA marker to describe the morphometry of T. cordata × platyphyllos hybrids of an impressive age (ca. 150 years) grown in the Pazaislis baroque monastery yard in Central Lithuania. In an earlier study on a country-wide set of 543 T. cordata individuals from natural forest populations in Lithuania, we detected a nuclear microsatellite locus Tc8 well-differentiating between T. cordata and T. platyphyllos. The Tc8 locus contained a 140 bp allele in T. cordata (541 sampled individuals) and alleles above 160 bp in the two trees with a T. platyphyllos-like morphology (sampled in a national park). To verify the Tc8 locus as species specific, we sampled a further four T. platyphyllos-like individuals, which all contained the Tc8 locus alleles above 160 bp. We subsequently genotyped the six old-growth individuals from the Pazaislis monastery with mixed T. cordata × platyphyllos morphology. Results revealed that all six old-growth Tilia individuals from the Pazaislis monastery were heterozygous for the Tc8 locus with alleles of 140 bp (indicative of T. cordata) and 162 bp (indicative of T. platyphyllos). This finding confirms the morphological observations that these individuals are hybrids between T. cordata and T. platyphyllos. Additionally, the genotyping of a set of 14 nuclear microsatellite loci revealed that all six trees from the Pazaislis monastery are clones, possessing identical microsatellite genotypes. After the molecular identification, we morphotyped leaves, bracts, twigs, and nuts of the 6 old-growth T. cordata × platyphyllos hybrids from the Pazaislis monastery, 16 T. cordata old-growth trees, 4 T. × europaea var. europaea 'Pallida' trees growing near the Pazaislis monastery, and 4 mature T. platyphyllos trees from a nearby Girionys park. The morphotyping showed that T. cordata × platyphyllos hybrids may be the easiest to distinguish from T. cordata by raised and horizontally tertiary veins of leaves.

Phenolic and Antioxidant Compound Accumulation of Quercus robur Bark Diverges Based on Tree Genotype, Phenology and Extraction Method.

Oak bark is a rich niche for beneficial bioactive compounds. It is known that the amount of the compounds found in plant tissues can depend on species, genotype, growth site, etc., but it is unclear whether oak phenology, i.e., late or early bud burst, can also influence the amount of phenols and antioxidants that can be extracted. We tested two Quercus robur populations expressing different phenology and five half-sib families in each population to see how phenology, genotype, as well as extrahent differences (75% methanol or water) can determine the total phenol, total flavonoid content, as well as antioxidant activity. Significant statistical differences were found between half-sib families of the same population, between populations representing different oak phenology and different extrahents used. We determined that the extraction of flavonoids was more favorable when using water. So was antioxidant activity using one of the indicators, when significant differences between extrahents were observed. Furthermore, in families where there was a significant difference, phenols showed better results when using methanol. Overall, late bud burst families exhibited higher levels in all parameters tested. Thus, we recommend that for further bioactive compound extraction, all these factors be noted.

Changes in Content of Bioactive Compounds and Antioxidant Activity Induced in Needles of Different Half-Sib Families of Norway Spruce (Picea abies (L.) H. Karst) by Seed Treatment with Cold Plasma.

In order to ensure sufficient food resources for a constantly growing human population, new technologies (e.g., cold plasma technologies) are being developed for increasing the germination and seedling growth without negative effects on the environment. Pinaceae species are considered a natural source of antioxidant compounds and are valued for their pharmaceutical and nutraceutical properties. In this study, the seeds of seven different Norway spruce half-sib families were processed for one or two minutes with cold plasma (CP) using dielectric barrier discharge (DBD) plasma equipment. At the end of the second vegetation season, the total flavonoid content (TFC), DPPH (2,2- diphenyl-1-picryl-hydrazyl-hydrate), and ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) antioxidant activity, and the amounts of six organic acids (folic, malic, citric, oxalic, succinic, and ascorbic) were determined in the needles of different half-sib families of Norway spruce seedlings. The results show that the TFC, antioxidant activity, and amounts of organic acids in the seedling needles depended on both the treatment duration and the genetic family. The strongest positive effect on the TFC was determined in the seedlings of the 477, 599, and 541 half-sib families after seed treatment with CP for 1 min (CP1). The TFC in these families increased from 118.06 mg g-1 to 312.6 mg g-1 compared to the control. Moreover, seed treatment with CP1 resulted in the strongest increase in the antioxidant activity of the needles of the 541 half-sib family seedlings; the antioxidant activity, determined by DPPH and ABTS tests, increased by 30 and 23%, respectively, compared to the control. The obtained results indicate that the CP effect on the amount of organic acids in the needles was dependent on the half-sib family. It was determined that treatment with CP1 increased the amount of five organic acids in the needles of the 541 half-sib family seedlings. The presented results show future possibilities for using cold plasma seed treatment in the food and pharmacy industries.

Genetic signature of the natural gene pool of Tilia cordata Mill. in Lithuania: Compound evolutionary and anthropogenic effects.

Tilia cordata Mill. is a valuable tree species enriching the ecological values of the coniferous-dominated boreal forests in Europe. Following the historical decline, spreading of Tilia sp. is challenged by the elevated inbreeding and habitat fragmentation. We studied the geographical distribution of genetic diversity of Tilia cordata populations in Lithuania. We used 14 genomic microsatellite markers to genotype 543 individuals from 23 wild-growing populations. We found that Tilia cordata retained high levels of genetic diversity (population F IS = 0-0.15, H o = 0.53-0.69, H e = 0.56-0.75). AMOVA, Bayesian clustering, and Monmonier's barrier detection indicate weak but significant differentiation among the populations (F ST = 0.037***) into geographically interpretable clusters of (a) western Lithuania with high genetic heterogeneity but low genetic diversity, bottleneck effects, (b) relatively higher genetic diversity of Tilia cordata on rich and most soils of midland lowland, and (c) the most differentiated populations on poor soils of the coolest northeastern highland possessing the highest rare allele frequency but elevated inbreeding and bottleneck effects. Weak genetic differentiation among the Tilia cordata populations in Lithuania implies common ancestry, absence of strong adaptive gradients, and effective genetic exchange possible mediated via the riparian networks. A hypothesis on riparian networks as gene flow mediators in Tilia cordata was raised based on results of this study.

Effect of Extracts from Dominant Forest Floor Species of Clear-Cuts on the Regeneration and Initial Growth of Pinus sylvestris L. with Respect to Climate Change.

Climate change influences the ecological environment and affects the recruitment of plants, in addition to population dynamics, including Scots pine regeneration processes. Therefore, the impact of cover-dominant species extracts on the germination of pine seeds and morpho-physiological traits of seedling under different environmental conditions was evaluated. Increasing temperature reinforces the plant-donor allelochemical effect, reduces Scots pine seed germination, and inhibits seedling morpho-physiological parameters. Conditions unfavourable for the seed germination rate were observed in response to the effect of aqueous extracts of 2-year-old Vaccinium vitis-ideae and 1-year-old Calluna vulgaris under changing environmental conditions. The lowest radicle length and hypocotyl growth were observed in response to the effect of 1-year-old C. vulgaris and 2-year-old Rumex acetosella under increasing temperature (+4 °C) conditions. The chlorophyll a + b concentration in control seedlings strongly decreased from 0.76 to 0.66 mg g-1 (due to current environmental and changing environmental conditions). These factors may reduce the resistance of Scots pine to the effects of dominant species and affect the migration of Scots pine habitats to more favourable environmental conditions.

Successional Categorization of European Hemi-boreal Forest Tree Species.

Developing forest harvesting regimes that mimic natural forest dynamics requires knowledge on typical species behaviors and how they respond to environmental conditions. Species regeneration and survival after disturbance depends on a species' life history traits. Therefore, forest succession determines the extent to which forest communities are able to cope with environmental change. The aim of this review was to (i) review the life history dynamics of hemi-boreal tree species in the context of ecological succession, and (ii) categorize each of these tree species into one of four successional development groups (gap colonizers, gap competitors, forest colonizers, or forest competitors). To do this we embraced the super-organism approach to plant communities using their life history dynamics and traits. Our review touches on the importance and vulnerability of these four types of successional groups, their absence and presence in the community, and how they can be used as a core component to evaluate if the development of the community is progressing towards the restoration of the climatic climax. Applying a theoretical framework to generate ideas, we suggest that forests should be managed to maintain environmental conditions that support the natural variety and sequence of tree species' life histories by promoting genetic invariance and to help secure ecosystem resilience for the future. This could be achieved by employing harvesting methods that emulate natural disturbances and regeneration programs that contribute to maintenance of the four successional groups.

Phenogenetic response of silver birch populations and half-sib families to elevated ozone and ultraviolet-B radiation at juvenile age.

Phenogenetic response of silver birch populations and half-sib families to separate and combined elevated ozone (O(3)) concentrations and ultraviolet-B (UV-B) radiation dozes was studied at juvenile age in the climatic chambers. Significant population and family effects were found for seedling height, lamina width, and leaf damage. The exposure to UV-B radiation decreased genetic variation at the stage of seed germination. Complex exposure to UV-B and O(3) caused an increase of genetic variation at the stage of intensive seedling growth: seedling height genetic variation in separate treatments increased from 23.7-38.6 to 33.7-65.7%, the increase for lamina width was from 10.2-13.9 to 13.6-31.8%. Different populations and families demonstrated differing response to elevated complex UV-B and O(3) exposure. Changes of genetic intra-population variation were population-specific. Such changes in genetic variation under the impact of stressors can alter adaptation, stability, and competitive ability of regenerating populations in a hardly predictive way.