Changes in Watering Frequency Stimulate Differentiated Adaptive Responses among Seedlings of Different Beech Populations.

Seasonality, rather than annual precipitation levels, is expected to affect the adaptive responses of plant populations under future climate change. To estimate adaptive traits' variation, we conducted a common garden experiment with two beech populations from contrasting climatic origins (Evros with longer drought intervals during summer and higher precipitation seasonality, and Drama representing a more temperate ecosystem). We simulated two different watering treatments (frequent vs. non-frequent) on beech seedlings, according to predicted monthly precipitation levels expected to prevail in 2050 by the CSIRO MK3.6 SRESA1B model, considering as reference area a natural beech stand in Mt. Rodopi, Greece. A series of morphological and stem anatomical traits were measured. Seedling survival was greater for the Evros population compared to that of Drama under non-frequent watering, while no difference in survival was detected under frequent watering. Leaf morphological traits were not generally affected by watering frequency except for leaf circularity, which was found to be lower under non-frequent watering for both populations. Stomata density in leaves was found to be higher in the Evros population and lower in the Drama population under non-frequent watering than frequent. Stem anatomical traits were higher under non-frequent watering for Evros but lower for the Drama population. Multivariate analyses clearly discriminated populations under non-frequent rather than frequent watering, indicating genetic adaptation to the population's environment of origin.

5S-IGS rDNA in wind-pollinated trees (Fagus L.) encapsulates 55 million years of reticulate evolution and hybrid origins of modern species.

Standard models of plant speciation assume strictly dichotomous genealogies in which a species, the ancestor, is replaced by two offspring species. The reality in wind-pollinated trees with long evolutionary histories is more complex: species evolve from other species through isolation when genetic drift exceeds gene flow; lineage mixing can give rise to new species (hybrid taxa such as nothospecies and allopolyploids). The multi-copy, potentially multi-locus 5S rDNA is one of few gene regions conserving signal from dichotomous and reticulate evolutionary processes down to the level of intra-genomic recombination. Therefore, it can provide unique insights into the dynamic speciation processes of lineages that diversified tens of millions of years ago. Here, we provide the first high-throughput sequencing (HTS) of the 5S intergenic spacers (5S-IGS) for a lineage of wind-pollinated subtropical to temperate trees, the Fagus crenata - F. sylvatica s.l. lineage, and its distant relative F. japonica. The observed 4963 unique 5S-IGS variants reflect a complex history of hybrid origins, lineage sorting, mixing via secondary gene flow, and intra-genomic competition between two or more paralogous-homoeologous 5S rDNA lineages. We show that modern species are genetic mosaics and represent a striking case of ongoing reticulate evolution during the past 55 million years.

Whole-Genome Sequencing, Phylogenetic and Genomic Analysis of Lactiplantibacillus pentosus L33, a Potential Probiotic Strain Isolated From Fermented Sausages.

Lactobacillus is a diverse genus that includes species of industrial and biomedical interest. Lactiplantibacillus pentosus, formerly known as Lactobacillus pentosus, is a recently reclassified species, that contains strains isolated from diverse environmental niches, ranging from fermented products to mammalian gut microbiota. Importantly, several L. pentosus strains present health-promoting properties, such as immunomodulatory and antiproliferative activities, and are regarded as potential probiotic strains. In this study, we present the draft genome sequence of the potential probiotic strain L. pentosus L33, originally isolated from fermented sausages. Comprehensive bioinformatic analysis and whole-genome annotation were performed to highlight the genetic loci involved in host-microbe interactions and the probiotic phenotype. Consequently, we found that this strain codes for bile salt hydrolases, adhesins and moonlighting proteins, and for Class IIb bacteriocin peptides lacking the GxxxG and GxxxG-like motifs, crucial for their inhibitory activity. Its adhesion ability was also validated in vitro, on human cancer cells. Furthermore, L. pentosus L33 contains an exopolysaccharide (EPS) biosynthesis cluster, and it does not carry transferable antibiotic resistance genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and CAZymes analyses showed that L. pentosus L33 possesses biosynthetic pathways for seven amino acids, while it can degrade a wide array of carbohydrates. In parallel, Clusters of Orthologous Groups (COGs) and KEGG profiles of L. pentosus L33 are similar to those of 26 L. pentosus strains, as well as of two well documented L. plantarum probiotic strains. Conclusively, L. pentosus L33 exhibits good probiotic potential, although further studies are needed to elucidate the extent of its biological properties.

Genomic and Phylogenetic Analysis of Lactiplantibacillus plantarum L125, and Evaluation of Its Anti-Proliferative and Cytotoxic Activity in Cancer Cells.

Lactiplantibacillus plantarum is a diverse species that includes nomadic strains isolated from a variety of environmental niches. Several L. plantarum strains are being incorporated in fermented foodstuffs as starter cultures, while some of them have also been characterized as probiotics. In this study, we present the draft genome sequence of L. plantarum L125, a potential probiotic strain presenting biotechnological interest, originally isolated from a traditional fermented meat product. Phylogenetic and comparative genomic analysis with other potential probiotic L. plantarum strains were performed to determine its evolutionary relationships. Furthermore, we located genes involved in the probiotic phenotype by whole genome annotation. Indeed, genes coding for proteins mediating host-microbe interactions and bile salt, heat and cold stress tolerance were identified. Concerning the potential health-promoting attributes of the novel strain, we determined that L. plantarum L125 carries an incomplete plantaricin gene cluster, in agreement with previous in vitro findings, where no bacteriocin-like activity was detected. Moreover, we showed that cell-free culture supernatant (CFCS) of L. plantarum L125 exerts anti-proliferative, anti-clonogenic and anti-migration activity against the human colon adenocarcinoma cell line, HT-29. Conclusively, L. plantarum L125 presents desirable probiotic traits. Future studies will elucidate further its biological and health-related properties.

Adaptive Diversity of Beech Seedlings Under Climate Change Scenarios.

The ability of beech (Fagus sylvatica L.) populations to adapt to the ongoing climate change is especially important in the southern part of Europe, where environmental change is expected to be more intense. In this study, we tested the existing adaptive potential of eight beech populations from two provenances in N.E. Greece (Evros and Drama) that show differences in their environmental conditions and biogeographical background. Seedling survival, growth and leaf phenological traits were selected as adaptive traits and were measured under simulated controlled climate change conditions in a growth chamber. Seedling survival was also tested under current conditions in the field. In the growth chamber, simulated conditions of temperature and precipitation for the year 2050 were applied for 3 years, under two different irrigation schemes, where the same amount of water was distributed either frequently (once every week) or non-frequently (once in 20 days). The results showed that beech seedlings were generally able to survive under climate change conditions and showed adaptive differences among provenances and populations. Furthermore, changes in the duration of the growing season of seedlings were recorded in the growth chamber, allowing them to avoid environmental stress and high selection pressure. Differences were observed between populations and provenances in terms of temporal distribution patterns of precipitation and temperature, rather than the average annual or monthly values of these measures. Additionally, different adaptive strategies appeared among beech seedlings when the same amount of water was distributed differently within each month. This indicates that the physiological response mechanisms of beech individuals are very complex and depend on several interacting parameters. For this reason, the choice of beech provenances for translocation and use in afforestation or reforestation projects should consider the small scale ecotypic diversity of the species and view multiple environmental and climatic parameters in connection to each other.

Distribution and physical traits of red wood ant mounds in a managed Rhodope mountains forest.

Red wood ants (RWA) are of great ecological importance for the forest ecosystem. Forestry practices, like clear-cutting, and trampling load, due to tourism, logging, and grazing stock, can greatly affect their colonies, disturbing their microhabitat. RWA in Greek forests have not been investigated so far. We herein report on the distribution and morphological traits of Formica lugubris mounds studied in Elatia forest (Rhodope mountains, Northern Greece), an all-aged managed mixed forest where selective logging practices are performed. Nearby vegetation, slope, canopy cover, shrub density, and distance from the nearest neighboring trees were also recorded. Mound density was shown to be much higher in this Greek forest compared to RWA mounds in other European-managed forests. Furthermore, we recorded a continuous nest establishment, despite forest management disturbances and trampling load. Our study suggests that single-tree selective forestry practices are essential for creating ideal microhabitats for the RWA and, therefore, for maintaining RWA populations.