The Oldest Bryophyte Herbarium Specimens from Central Europe, Collected by M. E. Boretius in 1717: Taxonomy, Nomenclature, Datation and Ethnopharmacology.

The WA Herbarium at the University of Warsaw houses a collection of plants created in 1717 by Matthew Ernest Boretius. They were gathered in former East Prussia, near Angerburg, now Wegorzewo (Poland). It is the oldest plant collection from this part of Europe. Boretius compiled the herbarium as a collection of all the surrounding plants, but their folk names (Polish and German) recorded in the herbarium confirm the ethnobiological or ethnopharmaceutical importance of some species. We identified bryophyte species and checked the accuracy of their original identifications recorded in the herbarium. We provided their Latin (scientific, pre-Linnaean) nomenclature together with German and Polish vernacular names. We contextualised this information within the history of the medicinal use of bryophytes around 1717, when the plant collection was created. We also investigated whether the specimens could have come from Northeastern Poland. Mosses and liverworts from the herbarium were identified nomenclaturally (by means of their original scientific polynomial names written on herbarium sheets) and taxonomically. The herbarium holds two species and one subspecies of liverwort and 27 species and one variety of moss. The accuracy of the original identifications was assessed, with a particular focus on the species considered medicinal at the time. We found that bryophytes were poorly known in the time of Boretius, which was the last period in bryology before the introduction of magnifying devices into this science (this crucial step was made by Dillenius in 1741). The vernacular names used in the herbarium were recorded for Marchantia polymorpha and Polytrichum commune-the only two species with confirmed medicinal use by the year 1717.

First Phytochemical Profiling and In-Vitro Antiprotozoal Activity of Essential Oil and Extract of Plagiochila porelloides.

Volatiles metabolites from the liverwort Plagiochila porelloides harvested in Corsica were investigated by chromatographic and spectroscopic methods. In addition to already reported constituents, three new compounds were isolated by preparative chromatography and their structures were elucidated by mass spectrometry (MS) and NMR experiments. Hence, an atypic aliphatic compound, named 1,2-dihydro-4,5-dehydronerolidol and two isomers, (E) and (Z), possessing an unusual humbertiane skeleton (called p-menth-1-en-3-[2-methylbut-1-enyl]-8-ol) are newly reported and fully characterized in this work. The in vitro antiprotozoal activity of essential oil and extract of P. porelloides against Trypanosoma brucei brucei and Leishmania mexicana mexicana and cytotoxicity were determined. Essential oil and Et2O extract showed a moderate activity against T. brucei with IC50 values: 2.03 and 5.18 µg/mL, respectively. It is noteworthy that only the essential oil showed a high selectivity (SI = 11.7). Diethyl oxide extract exhibited moderate anticancer (cancerous macrophage-like murine cells) activity and also cytotoxicity (human normal fibroblast) with IC50 values: 1.25 and 2.96 µg/mL, respectively.

Chemical Stimulation of Heterocyte Differentiation by the Feather Moss Hylocomium splendens: a Potential New Step in Plant-Cyanobacteria Symbioses.

Cyanobacteria associated with mosses play a key role in the nitrogen (N) cycle in unpolluted ecosystems. Mosses have been found to release molecules that induce morphophysiological changes in epiphytic cyanobionts. Nevertheless, the extent of moss influence on these microorganisms remains unknown. To evaluate how mosses or their metabolites influence N2 fixation rates by cyanobacteria, we assessed the nitrogenase activity, heterocyte frequency and biomass of a cyanobacterial strain isolated from the feather moss Hylocomium splendens and a non-symbiotic strain when they were either growing by themselves, together with H. splendens or exposed to H. splendens water, acetone, ethanol, or isopropanol extracts. The same cyanobacterial strains were added to another moss (Taxiphyllum barbieri) and a liverwort (Monosolenium tenerum) to assess if these bryophytes affect N2 fixation differently. Although no significant increases in nitrogenase activity by the cyanobacteria were observed when in contact with H. splendens shoots, both the symbiotic and non-symbiotic cyanobacteria increased nitrogenase activity as well as heterocyte frequency significantly upon exposure to H. splendens ethanol extracts. Contact with T. barbieri shoots, on the other hand, did lead to increases in nitrogenase activity, indicating low host-specificity to cyanobacterial activity. These findings suggest that H. splendens produces heterocyte-differentiating factors (HDFs) that are capable of stimulating cyanobacterial N2 fixation regardless of symbiotic competency. Based on previous knowledge about the chemical ecology and dynamics of moss-cyanobacteria interactions, we speculate that HDF expression by the host takes place in a hypothetical new step occurring after plant colonization and the repression of hormogonia.

New Oligomeric Dihydrochalcones in the Moss Polytrichum commune: Identification, Isolation, and Antioxidant Activity.

One of the most widespread representatives of mosses in the temperate and boreal latitudes of the Northern Hemisphere is common haircap (Polytrichum commune), which is known as the largest moss in the world and widely used in traditional herbal medicine. Polyphenolic compounds constitute one of the most important groups of biologically active secondary metabolites of P. commune, however, the available information on their chemical composition is still incomplete and contradictory. In the present study, a group of dihydrochalcone polyphenolic derivatives that were not previously found in mosses was isolated from P. commune biomass using pressurized liquid extraction with aqueous acetone. The combination of two-dimensional NMR spectroscopy and high-performance liquid chromatography-high-resolution mass spectrometry allowed for identifying them as 3-hydroxyphloretin oligomers formed through a carbon-carbon bond between phloroglucinol and pyrocatechol moieties ("head-to-tail" coupling), with a polymerization degree of 2-5. The individual compounds isolated by preparative reverse-phase HPLC had a purity of 71 to 97% and demonstrated high radical scavenging activity (17.5-42.5% with respect to Trolox) determined by the photochemiluminescence method. Along with the low toxicity predicted by QSAR/QSTR algorithms, this makes 3-hydroxyphloretin oligomers a promising source for the production of biologically active food additives and pharmaceuticals.

Green Christmas: bryophytes as ornamentals in Croatian traditional nativity scenes.

BACKGROUND: The bryophytes are a plant group that is smaller than and not as well known as the vascular plants. They are less used and are almost completely neglected in ethnobotanical studies. Traditional nativity scenes depicting the birth of Christ are commonly decorated with both vascular plants and bryophytes. The aim of this study was to document the diversity of decorative bryophytes sold during the Advent season at farmers' markets in Croatia (Southeastern Europe, Balkan Peninsula). METHODS: Twenty-eight farmers' markets in the two largest Croatian cities (Zagreb in the continental part and Split in the Mediterranean part) were studied in the search for local vendors selling bryophytes during the pre-Christmas season. The bryophytes collected were identified and analysed with respect to families, growth type, life forms and threat status. RESULTS: Among 275 collected specimens, 43 moss and four liverwort species were identified. The mean number of species per vendor was 3.5. The most frequent species were Hypnum cupressiforme, Homalothecium sericeum and Ctenidium molluscum. Mats, wefts and tufts were the most common life-forms, while pleurocarpous prevailed over acrocarpous mosses, as they are usually pinnately branched and form large carpets, suitable for decorations. The overall selection of bryophytes and the decorations made of them were more diverse and abundant in inland Croatia, where 49 vendors at 15 farmers' markets sold goods containing 43 species. In Mediterranean Croatia, at six farmers' markets only 29 vendors sold goods, which contained 18 species. A considerable number of species that are less attractive to harvesters were collected non-intentionally, entangled in carpets of other, more frequent species. Among them, Rhodobryum ontariense and Loeskeobryum brevirostre are rare and insufficiently recorded in Croatia so far. CONCLUSIONS: The present study provided a first perspective on the use of bryophytes in traditional nativity scenes in Croatia and Southeastern Europe, contributing to scarce ethnobotanical documentation of the decorative use of bryophytes in Christmas festivities in Europe and globally.

Anti-Inflammatory Activity of Bryophytes Extracts in LPS-Stimulated RAW264.7 Murine Macrophages.

Bryophytes produce rare and bioactive compounds with a broad range of therapeutic potential, and many species are reported in ethnomedicinal uses. However, only a few studies have investigated their potential as natural anti-inflammatory drug candidate compounds. The present study investigates the anti-inflammatory effects of thirty-two species of bryophytes, including mosses and liverworts, on Raw 264.7 murine macrophages stimulated with lipopolysaccharide (LPS) or recombinant human peroxiredoxin (hPrx1). The 70% ethanol extracts of bryophytes were screened for their potential to reduce the production of nitric oxide (NO), an important pro-inflammatory mediator. Among the analyzed extracts, two moss species significantly inhibited LPS-induced NO production without cytotoxic effects. The bioactive extracts of Dicranum majus and Thuidium delicatulum inhibited NO production in a concentration-dependent manner with IC50 values of 1.04 and 1.54 µg/mL, respectively. The crude 70% ethanol and ethyl acetate extracts were then partitioned with different solvents in increasing order of polarity (n-hexane, diethyl ether, chloroform, ethyl acetate, and n-butanol). The fractions were screened for their inhibitory effects on NO production stimulated with LPS at 1 ng/mL or 10 ng/mL. The NO production levels were significantly affected by the fractions of decreasing polarity such as n-hexane and diethyl ether ones. Therefore, the potential of these extracts to inhibit the LPS-induced NO pathway suggests their effective properties in attenuating inflammation and could represent a perspective for the development of innovative therapeutic agents.

The complete chloroplast genome of Rhodobryum laxelimbatum (Hampe ex Ochi) Z. Iwatsuki and T. J. Koponen.

Rhodobryum laxelimbatum (Bryophyta, Bryaceae) is one of the folk medicine resources in Southwest China, which has excellent potential for application in treating cardiovascular diseases. In this study, R. laxelimbatum was sequenced by high-throughput sequencing technology. The complete chloroplast genome is 124,632 bp in length with a quadripartite structure. Two inverted repeat regions are 9837 bp, separated by a large single copy region of 86,444 bp and a small single copy region of 18,514 bp. It encodes 118 unique genes, including 82 protein-coding genes, 32 tRNA genes, and four rRNA genes. The phylogenetic tree was constructed based on the complete chloroplast genome sequences of 18 bryophytes, downloaded from GenBank and acquired in this study. The phylogenetic analysis strongly indicated that R. laxelimbatum was the sister group of the clade which consists of Mnium marginatum, Pohlia cruda and Pohlia nutans. The R. laxelimbatum chloroplast genome sequence provides new genomic resources, which will improve its research, conservation, and application in the future.

Bryo-Activities: A Review on How Bryophytes Are Contributing to the Arsenal of Natural Bioactive Compounds against Fungi.

Usually regarded as less evolved than their more recently diverged vascular sisters, which currently dominate vegetation landscape, bryophytes seem having nothing to envy to the defensive arsenal of other plants, since they had acquired a suite of chemical traits that allowed them to adapt and persist on land. In fact, these closest modern relatives of the ancestors to the earliest terrestrial plants proved to be marvelous chemists, as they traditionally were a popular remedy among tribal people all over the world, that exploit their pharmacological properties to cure the most different diseases. The phytochemistry of bryophytes exhibits a stunning assortment of biologically active compounds such as lipids, proteins, steroids, organic acids, alcohols, aliphatic and aromatic compounds, polyphenols, terpenoids, acetogenins and phenylquinones, thus it is not surprising that substances obtained from various species belonging to such ancestral plants are widely employed as antitumor, antipyretic, insecticidal and antimicrobial. This review explores in particular the antifungal potential of the three Bryophyta divisions-mosses (Musci), hornworts (Anthocerotae) and liverworts (Hepaticae)-to be used as a sources of interesting bioactive constituents for both pharmaceutical and agricultural areas, providing an updated overview of the latest relevant insights.

Water Extract of Cryphaea heteromalla (Hedw.) D. Mohr Bryophyte as a Natural Powerful Source of Biologically Active Compounds.

Bryophytes comprise of the mosses, liverworts, and hornworts. Cryphaea heteromalla, (Hedw.) D. Mohr, is a non-vascular lower plant belonging to mosses group. To the date, the most chemically characterized species belong to the liverworts, while only 3.2% and 8.8% of the species belonging to the mosses and hornworts, respectively, have been investigated. In this work, we present Folin-Ciocalteu and oxygen radical absorbance capacity (ORAC) data related to crude extracts of C. heteromalla obtained by three different extraction solvents: pure water (WT), methanol:water (80:20 v/v) (MET), and ethanol:water (80:20 v/v) (ETH). The water extract proved to be the best solvent showing the highest content of biophenols and the highest ORAC value. The C. heteromalla-WT extract was investigated by HPLC-TOF/MS (High Performance Liquid Chromatography-Time of Flight/Mass Spectrometry) allowing for the detection of 14 compounds, five of which were phenolic compounds, derivatives of benzoic, caffeic, and coumaric acids. Moreover, the C. heteromalla WT extract showed a protective effect against reactive oxygen species (ROS) generation induced by tert-butyl hydroperoxide (TBH) on the murine NIH-3T3 fibroblast cell line.

Antiproliferative and Antimicrobial Activities of Selected Bryophytes.

One-hundred and sixty-eight aqueous and organic extracts of 42 selected bryophyte species were screened in vitro for antiproliferative activity on a panel of human gynecological cancer cell lines containing HeLa (cervix epithelial adenocarcinoma), A2780 (ovarian carcinoma), and T47D (invasive ductal breast carcinoma) cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and for antibacterial activity on 11 strains using the disc-diffusion method. A total of 99 extracts derived from 41 species exerted ≥25% inhibition of proliferation of at least one of the cancer cell lines at 10 µg/mL. In the cases of Brachythecium rutabulum, Encalypta streptocarpa, Climacium dendroides, Neckera besseri, Pleurozium schreberi, and Pseudoleskeella nervosa, more than one extract was active in the antiproliferative assay, whereas the highest activity was observed in the case of Paraleucobryum longifolium. From the tested families, Brachytheciaceae and Amblystegiaceae provided the highest number of antiproliferative extracts. Only 19 samples of 15 taxa showed moderate antibacterial activity, including the most active Plagiomnium cuspidatum, being active on 8 tested strains. Methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus were the most susceptible to the assayed species. This is the first report on the bioactivities of these 14 species.

Lichen secondary metabolites affect growth of Physcomitrella patens by allelopathy.

Lichen secondary metabolites can function as allelochemicals and affect the development and growth of neighboring bryophytes, fungi, vascular plants, microorganisms, and even other lichens. Lichen overgrowth on bryophytes is frequently observed in nature even though mosses grow faster than lichens, but there is still little information on the interactions between lichens and bryophytes.In the present study, we used extracts from six lichen thalli containing secondary metabolites like usnic acid, protocetraric acid, atranorin, lecanoric acid, nortistic acid, and thamnolic acid. To observe the influence of these metabolites on bryophytes, the moss Physcomitrella patens was cultivated for 5 weeks under laboratory conditions and treated with lichen extracts. Toxicity of natural mixtures of secondary metabolites was tested at three selected doses (0.001, 0.01, and 0.1 %). When the mixture contained substantial amounts of usnic acid, we observed growth inhibition of protonemata and reduced development of gametophores. Significant differences in cell lengths and widths were also noticed. Furthermore, usnic acid had a strong effect on cell division in protonemata suggesting a strong impact on the early stages of bryophyte development by allelochemicals contained in the lichen secondary metabolites.Biological activities of lichen secondary metabolites were confirmed in several studies such as antiviral, antibacterial, antitumor, antiherbivore, antioxidant, antipyretic, and analgetic action or photoprotection. This work aimed to expand the knowledge on allelopathic effects on bryophyte growth.

Central European medicinal bryophytes in the 16th-century work by Caspar Schwenckfeld, and their ethnopharmacological origin.

ETHNOPHARMACOLOGICAL RELEVANCE: Ubiquitous bryophyte species are reported from European, American and Asian ethnopharmacies. Some of their traditional medicinal uses are similar in distant and isolated cultures, and moreover, medicinal properties of some bryophytes are currently confirmed as justified by their chemical constituents. Aims of the work: we identify bryophytes listed in a medicinal and botanical work from 1600, and compare their medicinal applications (known in Europe between 1530-1600) with other ethnopharmacological data about these species and with modern pharmacological knowledge. This way we attempt to display origins of medicinal usage of bryophytes in Central Europe. MATERIALS AND METHODS: Bryophyte species in bibliographical sources printed in Central Europe (starting from O. Brunfels' Herbarum vivae Eicones... Argentorati, 1530) were identified according to old and recent taxonomical references. Caspar Schwenckfeld's scientific output from 1600 was treated here as a summary of 16th-century knowledge about medicinal bryophytes. RESULTS: Central European pharmacy about the year 1600 was familiar with the following bryophytes: Marchantia polymorpha L., Polytrichum commune Hedw., P. formosum Hedw. and Funaria hygrometrica Hedw. CONCLUSIONS: Between 1530-1600 in Central Europe the number of medicinal bryophytes increased from 2 (Lichen sive Hepatica and Polytrichon) to 4. Pharmaceutical usage of them was similar as in other, distant ethnopharmacies. Further 2-4 mosses (Rhytidiadelphus loreus and Rh. squarrosus; Thuidium tamariscinum and Th. delicatulum) were recognised as non-medicinal.

Slow recovery of High Arctic heath communities from nitrogen enrichment.

Arctic ecosystems are strongly nutrient limited and exhibit dramatic responses to nitrogen (N) enrichment, the reversibility of which is unknown. This study uniquely assesses the potential for tundra heath to recover from N deposition and the influence of phosphorus (P) availability on recovery. We revisited an experiment in Svalbard, established in 1991, in which N was applied at rates representing atmospheric N deposition in Europe (10 and 50 kg N ha(-1)  yr(-1) ; 'low' and 'high', respectively) for 3-8 yr. We investigated whether significant effects on vegetation composition and ecosystem nutrient status persisted up to 18 yr post-treatment. Although the tundra heath is no longer N saturated, N treatment effects persist and are strongly P-dependent. Vegetation was more resilient to N where no P was added, although shrub cover is still reduced in low-N plots. Where P was also added (5 kg P ha(-1)  yr(-1) ), there are still effects of low N on community composition and nutrient dynamics. High N, with and without P, has many lasting impacts. Importantly, N + P has caused dramatically increased moss abundance, which influences nutrient dynamics. Our key finding is that Arctic ecosystems are slow to recover from even small N inputs, particularly where P is not limiting.

Phytotoxic cis-clerodane diterpenoids from the Chinese liverwort Scapania stephanii.

Five cis-clerodane diterpenoids, stephanialides A-E, along with seven known cis-clerodanes, scaparvins A-C, parvitexins B and C, 3-chloro-4-hydroxy-parvitexin A, and scapanialide B, were isolated from the Chinese liverwort Scapania stephanii. Their structures were established unequivocally on the basis of spectroscopic data. The absolute configuration of stephanialide A was determined by analysis of CD data using the octant rule. Phytotoxic activity evaluation showed that this type of diterpenoids can significantly inhibit root elongation of the seeds of Arabidopsis thaliana, Lepidium sativum and Brassica pekinensis.

Medicinal mosses in pre-Linnaean bryophyte floras of central Europe. An example from the natural history of Poland.

ETHNOPHARMACOLOGICAL RELEVANCE: The paper presents information about the earliest botanical work from Poland, Warsavia physice illustrata which takes bryophytes into account. It was elaborated by a German physician Christian Heinrich Erndtel and issued in 1730 in Dresden. That time understanding of bryophytes was imprecise and in many cases they were confused with lichens and club mosses. MATERIALS AND METHODS: Bryophyte taxa polynomials (18 names) were identified using pre- and post-Linnaean botanical monographs from years 1590 to 1801. Their current names and pharmacological value are provided, as well as the old ethnobotanical data about bryophytes (cited from 18th-century sources). RESULTS: Altogether 18 bryophyte species were identified from the vicinity of Warsaw (17 mosses and 1 liverwort). Some of them are still abundant in this area (for example Climacium dendroides, Plagiomnium undulatum and Polytrichum juniperinum) while some other are rare or extinct (for example Neckera crispa and Rhodobryum roseum). CONCLUSIONS: Despite the technical ability to observe specific microscopic differences among bryophytes, physicians of 18th century were hardly interested in using any of them as medicinal stock. It may be concluded that the competences in pre-Linnaean bryology did not put into practice using moss-derived materia medica of 18th century (the only exceptions were Fontinalis antipyretica and Polytrichum spp.).