Abstract The minuscule but evolutionarily and environmentally important mosses are early land plants. The majority of moss plants have erect growth forms; however, how these tiny plants manage to stand upright has not been fully explored. The information on the type and location of tissues proposed to be responsible for bearing the load of the highly hydrated body parts comes mainly from microscopy studies. Based on light microscopy examination of stem sections, it has been proposed that thick-walled outer tissues are the likely candidates for this function. Recent studies employing transmission electron microscopy have shown that thickened cell walls consist of multiple lamellae, with alternating thick and thin lamellae differing in the orientation of cellulose microfibrils. This cell wall design optimizes mechanical properties by strengthening and supporting tissues in higher land plants and points to early evolution of this type of cell wall design, which likely occurred in mosses.

The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a widely used programmable nuclease system for gene modification in many organisms, including Physcomitrium patens. P. patens is a model species of moss plants, a basal land plant group, which has been extensively studied from the viewpoint of evolution and diversity of green plant lineages. So far, gene modifications by CRISPR/Cas9 in P. patens have been carried out exclusively by the polyethylene glycol (PEG)-mediated DNA transfer method, in which a transgene (or transgenes) is introduced into protoplast cells prepared from protonemal tissues. However, this PEG-mediated method requires a relatively large amount of transgene DNA (typically 30 µg for a single transformation), consists of many steps, and is time-consuming. Additionally, this PEG-mediated method has only been established in a few species of moss. In the current protocol, we succeeded in CRISPR/Cas9-induced targeted mutagenesis of P. patens genes by making good use of the biolistic method, which i) requires amounts of transgene DNA as low as 5 μg for each vector, ii) consists of fewer steps and is time-saving, and iii) is known to be applicable to a wide variety of species of plants.Key features• In this protocol, particle bombardment-mediated gene transfer is used for CRISPR/Cas9-induced mutagenesis in the moss Physcomitrium patens.• By this application of particle bombardment, a gene can be modified by the CRISPR/Cas9 system much more conveniently with a smaller amount of transgene DNA.• This protocol is expected to be easily applicable to non-model moss species, some of which have noteworthy traits, such as tolerance to various stresses.

First production of virus-like particles as a vaccine candidate in a non-vascular plant. Virus-like particles (VLPs) are self-assembling nanoparticles composed of viral structural proteins which mimic native virions but lack viral DNA and infectivity. VLPs are a resourceful class of biopharmaceuticals applied as subunit vaccines or as delivery vehicles for drugs and nucleic acids. Similar to viruses, VLPs are diverse in structure, composition, and assembly, requiring a tailored production platform aligned with the intended application. The moss plant Physcomitrella (Physcomitrium patens) is an emerging expression system offering humanized N-glycosylation, scalability, and adaptability to existing industry settings. Here, we used Physcomitrella to produce human papillomavirus (HPV) 16 VLPs. HPV VLPs are composed of the major structural protein L1 and are used as vaccines against HPV infections which are the main causal agent of cervical and other anogenital cancers. We characterized Physcomitrella chloroplast transit peptides, which we used for targeting of moss-produced L1 to chloroplasts, leading to higher recombinant protein yield compared to nuclear or cytoplasmic localization. We confirmed subcellular localization with confocal laser scanning microscopy and found L1 to accumulate within the chloroplast stroma. Production in 5-L photobioreactors yielded over 0.3 mg L1 per gram fresh weight. We established a purification protocol for moss-produced L1 using a combination of ammonium sulphate precipitation and cation exchange chromatography. Purified samples were subjected to a controlled dis- and reassembly, yielding fully assembled HPV-16 L1 VLPs. This is the first report of production, purification, and assembly of VLPs in a non-vascular plant.

To enhance the interfacial performance of Al/Mg bimetal, this study introduced a novel Mo coating and employed an ultrasonic field (UF) to regulate the interfacial microstructure. In the absence of both a Mo coating and ultrasonic treatment (referred to as the untreated specimen), the interfacial region was primarily composed of Al-Mg intermetallic compounds (Al-Mg IMCs), Al-Mg eutectic structures (ES), and Mg2Si phases, with an average interfacial layer thickness of approximately 1623 μm. Upon application of the Mo coating, the formation of both Al-Mg phases and Mg2Si phases was completely inhibited. The interfacial zone was predominantly characterized by the Mo solid solution (Mo SS) and oxide, with the average thickness significantly reduced to about 28 μm. Upon applying the UF to the Mo-coated specimen, the interfacial composition remained similar to that of the untreated specimen, except for Mo SS, with the interfacial thickness increasing to 561 μm. Shear strength tests indicated that the application of the Mo coating alone resulted in a decrease in bonding strength compared to the untreated specimen. However, subsequent ultrasonic treatment significantly improved the interfacial shear strength to 54.7 MPa, representing a 60.9% increase relative to the untreated specimen. This improvement is primarily attributed to the Mo coating and UF synergistically suppressing the formation of brittle Al-Mg IMCs and reducing oxide inclusions at the interface. Thus, the simultaneous application of Mo coatings and ultrasonic fields is required to enhance the properties of Al/Mg bimetals.

Abstract Many species of river riparia are threatened by habitat loss due to altered flood and sediment regime, and associated shifts in vegetation structure. However, their ecological niche is often obscure, especially in inconspicuous organisms such as lichens, hindering their conservation and use as indicator species in river restoration. We studied if variation in sediment size distribution, gravel bank elevation and vegetation structure drive presence-absence and fertility (fruit body production) in the endangered, soil-dwelling lichen S tereocaulon incrustatum along two Swiss braided rivers, using binomial generalized linear mixed effect models in a Bayesian framework. Data was sampled on 811 plots randomly placed along 41 transects perpendicular to the main channels. Presence probability was highest on the most elevated plots, at 30% vascular plant cover in the herb layer, and 30–40% cobble cover, and increased with moss cover. Fruit body production probability was highest under closed canopies of woody plants > 3m. We show that in braided rivers, S. incrustatum is most likely found on elevated, coarse-grained sediments with increased moss but moderate vascular plant cover. This indicates a niche comprising relatively stable riparian environments, where a dry, cryptogam-dominated vegetation establishes on raw soils and competition with vascular plants is moderate. Fertile thalli are mostly found under closed canopies and high densities, suggesting a shift to sexual reproduction with increasing habitat age. While rare but strong disturbances are therefore necessary for habitat creation, older, densely populated habitat patches may harbor important source populations for colonization, thereby representing focal areas for conservation.

ABSTRACT The cultivation and inoculation of lithophytic moss crusts represent a potential pathway for the ecological restoration of rocky slopes. At present, the key factors influencing rapid cultivation are not well understood, and ecological restoration practices remain at the conceptual stage. In this study, based on the investigation of moss diversity in the northern foothills of the Qinling Mountains, we selected Brachythecium plumosum , a dominant moss species, for incubator experiments to explore the inoculation density (20 g·m −2 , 60 g·m −2 , 100 g·m −2 ), moss stem and leaf fragment length (1–3 mm, 3–6 mm, 6–9 mm), and substrate type (kaguma soil, peat soil, volcanic rock) on moss crust growth. Meanwhile, an outdoor artificially constructed rocky slope was utilized to examine the response of moss crust restoration on rocky slopes to inoculation methods (dry broadcasting, wet blending), substrate particle sizes (< 1 mm, 1–2 mm, > 2 mm), gauze additions, and exogenous microorganisms ( Bacillus megaterium , Chlorella vulgaris ). The results showed that (1) in an artificial incubator (constant environment), medium inoculum, short stem and leaf fragments, and volcanic rock substrate were more conducive to the rapid development of moss crusts. Moss coverage, plant height, and plant density reached 50.28%, 5.64 mm, and 52 stems·cm −2 , respectively, in the 60 g·m 2 + 1–3 mm gametophyte fragments + volcanic rock treatment; (2) Moss coverage, plant height, and plant density of dry broadcast inoculation were significantly higher than those of wet blend inoculation, but the addition of B. megaterium and C. vulgaris did not produce significant differences; the addition of gauze cushion and substrate (particle size < 1 mm) contributed to rapid establishment and improved survival rates of moss crusts on rocky slopes. In summary, we identified the key factors for the rapid cultivation of lithophytic moss crust, preliminarily verified its feasibility as an ecological restoration measure for rocky slopes, and provided methodological references and new perspectives for the restoration of bare rock landscapes.

The emergence of land plants required adaptations to altered water availability and increased effective gravity. Bryophytes underwent major changes in physiology, anatomy, and growth during their emergence onto land. However, the link between gravity, photosynthesis, and genetic control remains unclear. Here, we show that leaf carbon dioxide diffusion enhanced photosynthesis in the model moss Physcomitrium patens under increased gravity (6 and 10 times Earth's gravity), driven by increases in plant (gametophore) number and chloroplast size. RNA sequencing analysis showed that 10 gravity up-regulated several species-specific APETALA2/ethylene-responsive factor (AP2/ERF) transcription factors. Overexpression of one such AP2/ERF, ISSUNBOSHI1 (IBSH1; gene ID=Pp3c1_32440), in P. patens phenocopied plants grown at 10 gravity, and the dominant repressor form of IBSH1 suppressed 10 gravity responses. These results provide evidence that the proliferation of AP2/ERF transcription factors and the establishment of a notable gene network may have been important in adaptation to the terrestrial environment during land plant evolution.

The restoration of moss crust (moss-dominated biological soil crust) through artificial cultivation is of great significance for preventing soil wind erosion and desertification in arid and semi-arid regions. However, the tiny size of moss plant and associated observational challenge hindered the establishment of scientifically validated and standardized indicators for assessing the growth and development of artificial moss crusts. We measured moss crust development indicators, including plant density, coverage, and height every five days during a 60-day cultivation experiment, as well as the dry weight and thickness of moss crust after the cultivation. We determined the eva-luation indicators for the artificial cultivation of moss crusts by analyzing the relationship between those indicators and the stability (characterization of erosion resistance) of moss crusts. The results showed that the new gametophytes germinated after five days cultivation. Plant individual density and coverage of moss crust increased rapidly and turned stable after 35 to 40 days of cultivation. The moss height remained within a narrow range of 1.53 to 1.63 mm during the cultivation. Dry weight of moss and thickness of moss crusts reached 0.0074 g·cm-2 and 6.30 mm at the end of cultivation, respectively. Moss density exhibited the highest sensitivity to cultivation time, followed by coverage, whereas plant height showed relatively low sensitivity. The stability index of moss crusts cultivated for 60 days ranged from 0.12 to 0.69, which were positively related with plant density and coverage. The comprehensive scoring for development indicators, including moss density, coverage, plant height, biomass, and thickness of moss crusts indicated that moss plant density and coverage are key indicators for assessing the growth and development of artificial moss crusts cultivation.

Bryophytes have gained recognition as an exceptional source of novel natural products, including secondary metabolites with intriguing biological activities. This study explored the antibacterial potential of bryophytes (Calymperes erosum and Racopilum africanum) as sources of novel natural products with significant biological activities. The antibacterial properties of these moss plants' powder and methanol extracts were evaluated against six bacterial strains using the agar-well diffusion and broth-dilution methods. Quantitative phytochemical analysis revealed the presence of various compounds, including tannins, saponins, flavonoids, glycosides, alkaloids, and phenols, with Calymperes erosum exhibiting higher levels of these phytochemicals. Remarkably, the bryophyte powder exhibited more significant antibacterial efficacy compared with the methanol extracts. Particularly, Calymperes erosum powder at 1000 mg/ml demonstrated the highest zone of inhibition against Escherichia coli (80 mm) and Salmonella typhi (32 mm). However, no inhibition was observed when both moss plant powders were tested against Plesiomonas shigelloides at various concentrations. The results were compared with standard antibiotic discs, and all test organisms displayed sensitivity to Racopilum africanum (MIC < 0.1 mg/ml) and Calymperes erosum (MIC 1 mg/ml). This study suggests the potential of Calymperes erosum and Racopilum africanum powders as sources for future antibacterial drug development, highlighting their value in pharmaceutical research.

Education is a fundamental pillar for individual development, and learning forms the core of this process. The act of learning, which essentially involves expanding horizons, deepening knowledge, and constructing changes in behavior, often faces various implementation challenges. Conventional teaching methods, such as the use of blackboards as the primary medium, are considered less effective in accommodating the dynamic learning styles of students. Traditional methods of teaching present several significant drawbacks, including a decline in student concentration while absorbing theoretical material presented by teachers, and the emergence of a monotonous and uninspiring learning environment that can hinder learning interest. These issues are particularly critical within the realm of biology education, especially regarding topics like moss plants that tend to be abstract and require effective visualization. In the midst of these challenges, gamification appears as an innovative solution. Gamification, which utilizes game elements in a non-game context, has been proven to effectively boost student learning motivation. This approach transforms the learning experience from being tedious to something more fun and engaging. Through the utilization of technology, gamification presents challenging material in a more digestible format, which also results in an interactive learning experience. This research aims to develop a gamified learning application that is attractive, convenient, practical, and engaging. This application makes use of QR codes to provide access to subtopics in biology, particularly moss plants. The gamified learning system applies a reward mechanism to increase student appeal. Each QR code leads students to different subtopics, which in turn create a dynamic learning path. The research methodology comprises: (1) analysis of the research context, (2) identification of participants (junior high school students), (3) design of a gamified learning model, and (4) assessment of learning motivation. Data were collected through in-depth interviews and questionnaires from 17 junior high school students, aged 12-14. The interviews explore student learning experiences, while questionnaires measure learning motivation before and after application usage. This research integrates the KAHOOT! platform for learning evaluation, which presents evaluations in an interactive multimedia format. The results of this research indicate that learning via gamification approaches, alongside the integration of KAHOOT!, positively impacts the increase of student learning motivation. These findings suggest that gamification has significant potential as an effective and engaging alternative teaching method within the context of modern education.

Education is a fundamental pillar for individual development, and learning forms the core of this process. The act of learning, which essentially involves expanding horizons, deepening knowledge, and constructing changes in behavior, often faces various implementation challenges. Conventional teaching methods, such as the use of blackboards as the primary medium, are considered less effective in accommodating the dynamic learning styles of students. Traditional methods of teaching present several significant drawbacks, including a decline in student concentration while absorbing theoretical material presented by teachers, and the emergence of a monotonous and uninspiring learning environment that can hinder learning interest. These issues are particularly critical within the realm of biology education, especially regarding topics like moss plants that tend to be abstract and require effective visualization. In the midst of these challenges, gamification appears as an innovative solution. Gamification, which utilizes game elements in a non-game context, has been proven to effectively boost student learning motivation. This approach transforms the learning experience from being tedious to something more fun and engaging. Through the utilization of technology, gamification presents challenging material in a more digestible format, which also results in an interactive learning experience. This research aims to develop a gamified learning application that is attractive, convenient, practical, and engaging. This application makes use of QR codes to provide access to subtopics in biology, particularly moss plants. The gamified learning system applies a reward mechanism to increase student appeal. Each QR code leads students to different subtopics, which in turn create a dynamic learning path. The research methodology comprises: (1) analysis of the research context, (2) identification of participants (junior high school students), (3) design of a gamified learning model, and (4) assessment of learning motivation. Data were collected through in-depth interviews and questionnaires from 17 junior high school students, aged 12-14. The interviews explore student learning experiences, while questionnaires measure learning motivation before and after application usage. This research integrates the KAHOOT! platform for learning evaluation, which presents evaluations in an interactive multimedia format. The results of this research indicate that learning via gamification approaches, alongside the integration of KAHOOT!, positively impacts the increase of student learning motivation. These findings suggest that gamification has significant potential as an effective and engaging alternative teaching method within the context of modern education.

Mosses (Bryophyta) are lower plants that lack the vascular tissue, roots, stems and true leaves of higher plants. Lichens can grow in a variety of habitats, including in extreme environments. This study aims to identify the diversity of moss plants in Coban Talun, Batu City, East Java. The sampling technique applied in this study involved dividing the area into 10 plots, with each plot having a size of 3m x 3m. The results obtained from ten plots in the Coban Talun waterfall area showed the presence of 9 species of lichens growing in the location, namely Anoectangium aestivum, Anomodon attenuatus, Campylopus introflexus, Marchantia polymorpha, L. cruciata, Pogonatum inflexum, Andreaea sp., Leucobryum glaucum, and Conocephalum conicum. Moss diversity is strongly influenced by environmental conditions such as temperature, air humidity, and light intensity. At low temperatures (around 25°C) moss diversity is higher than at higher temperatures (around 29°C). At humidity close to 72%, liverwort diversity is more dominant, while at humidity around 62%, leafwort diversity is more dominant.

ABSTRACTDiscomfort caused by inflammation leads to stress and anxiety in patients and seriously decreases the patients' quality of life. People prefer to use natural products instead of anti‐inflammatory drugs because of their low toxicity and side effects. Studies have shown that Sphagnum palustre L. (S. palustre) can be used as medicinal plant, but few studies have focused on its anti‐inflammatory effects. This study explored the mechanism of action of the ethanol extract of the peat moss plant S. palustre on lipopolysaccharide‐induced inflammation in macrophage RAW264.7 cells. Components in S. palustre ethanol extracts (SPE) were identified by HPLC‐MS, which mainly included 4‐methoxybenzaldehyde, 4‐methoxycinnamaldehyde and oleanolic acid. The effects of different concentrations (6.25–100 μg/mL) of SPE after 24 h administration were evaluated to establish a cellular inflammation model. Three biological replicates were performed based on each experiment, the MTT assay results showed that a low concentration of SPE promoted cell proliferation marked by Formazan. In a neutral red uptake assay, the SPE group was effectively inhibited the cell phagocytosis rate. With the increase of SPE concentration, intracellular ROS release decreased, which detected by DCFH‐DA. Immunofluorescence assay result showed that SPE inhibited the release of reactive oxygen species from macrophages with fluorescent markers and DAPI. SPE inhibited the release of nitric oxide from macrophages as well. What's more, SPE significantly decreased the protein expression of interleukin (IL)‐1, IL‐6, and nuclear factor (NF)‐κB according to enzyme‐linked immunosorbent and immunocytochemical assays. SPE reduces inflammation in macrophage RAW264.7 cells and thus is a promising natural anti‐inflammatory plant.

Moss plants can live well in areas that have high humidity. The Dutch Guesthouse which is located in the Sultan Adam Mandiangin Forest Park area has high humidity because the area has lots of dense plants, so it is a suitable habitat for the living nature of moss plants. This study aims to describe and identify the various species of moss found in the area of study. Data collection was carried out using the exploring method, followed by determining and identifying each sample of the moss plants found in the laboratory. Data analysis was carried out descriptively and identifying species with reference to the relevant literatures. The results found 10 species of mosses belonging to 7 different families, both found in tree, rock and soil habitats. The species found were: Calymperes tenerum, Calymperes afzelii, Hyophila apiculata, Hyophila involuta, Ectropothecium falciforme, Octoblepharum albidum, Philonotis hastata, Acroporium secundum, Lejeunea lamacerina, and Thysananthus sp. The results of this study are expected to provide information about the variety and biodiversity of mosses in South Kalimantan.

Distinct lignocelluloses of plant evolution are optimally selective for complete biomass saccharification and upgrading Cd2+/Pb2+ and dye adsorption via desired biosorbent assembly

Bryophytes are a group of low-level plants that grow widely and are found in tropical areas. Their distribution is influenced by biotic and abiotic factors such as water, light, temperature, humidity, and altitude. The ecological role of lichens in tropical ecosystems is to maintain water balance, nutrient cycling, bioindicators of environmental change, open space for other plants, and carbon sinks on peatlands. The role of moss is very important for the ecosystem, and there is no data on moss species in the Silawe waterfall area, so researchers conducted this study to identify environmental factors that affect moss diversity. Data collection for moss plants was carried out through a combination of exploration and field observation methods. Based on the results of observations, 7 species of moss, 5 leaf mosses (Bryopsida), and 2 liverworts (Marchantiopsida) were obtained. The identified ferns came from 7 different families, namely Funariaceae, Dicranaceae, Fissidentaceae, Hypnaceae, Entodontaceae, Marchantiaceae, and Lepidoziaceae. The identified moss species were Funaria hygrometrica, Dicranum fuscescens, Fissidens sp., Hypnum cupressiforme, Entodon sp., Marchantia polymorpha, and Bazzania sp.

Two-dimensional crystals are a class of materials in which an individual layer is indefinitely extended in two dimensions. Each layer is the molecular unit in the crystalline lattice. The nature of interlayer/intermolecular interaction is poorly understood. However, it defines physical properties such as carrier mobility, thermal conductivity, photoluminescence, etc. The vibrational properties of 2D materials are described based on triperiodic space groups. However, the triperiodic space groups do not apply to the description of these structures. The 17 two-dimensional groups in triperiodic space groups do not allow the existence of a third dimension. Therefore, if structures are not planar, the approach is inappropriate. The way to describe such structures is using the diperiodic groups in three dimensions. The well-known 2D materials are graphite, graphene, chalcogenide of Ga, Mo, W, and other elements. The crystalline As2S3 belongs to a class of 2D materials. The unit cell contains two layers with ten atoms in each layer, and it is similar to a structure of metal chalcogenide (MoS(Se)2, WS(Se)2). A bond length of 2.24 A is within the layer, and 3.56 A is between the layers. This interlayer/intralayer bond length ratio (= 1.59) corresponds to a significant difference in bond strengths. Center-zone optical phonons in crystalline As2S3 have been investigated by Raman scattering in a wide temperature range for two polarizations in the layer plane (ac). The crystal's and individual layers' symmetry are different, defining different selection rules for optical phonons. According to the crystallographic data, crystal symmetry is monoclinic with 20 atoms in a unit cell and is P21/n (C2h 5). The axis b, perpendicular to the layers, is the only unique axis of the crystal. Therefore, all phonons in the ac plane should be indistinguishable. The layer symmetry is orthorhombic in the diperiodic space group in three dimensions, in which a, b, and c axes are axes of symmetry with a space group of Pnm21 (C2v 7). The intensity of Raman bands in the layer plane shows strong polarization dependence, indicating layer symmetry's dominance. Interlayer interaction is weak, and as a result, low-frequency Raman bands appear in the spectra. The position of these bands defines the strength of interlayer interaction. The data presented provides an understanding of the structural motives of 2D As2S3 and may predict the optical/electronic properties of similar 2D materials based on Raman spectra.

Non-vascular plants, such as mosses (Bryophyta), are commonly found on land. The class of plants known as tuberous plants, or plants that do not have true roots, stems, or leaves as well as xylem and phloem vessels, includes moss plants. Knowing the diversity of moss plants in Namorambe Sub-district is the purpose of this research. On May 9, 2024, this research was conducted by walking around the Namorambe area to look for moss plants that might be there. This research methodology is qualitative, using descriptive and exploratory techniques. This research used observation, documentation, and literature study as data collection methods. Direct investigation into the Namorambe area was used to make observations to identify moss plants. The three species identified based on the results of the identification of leaf mosses around Namorambe Sub-district were Hyophila sp., Pogonatum cirrhatum, and Barbula indica. Leaf mosses were made near people's work areas and were made on tree bark and stone substrates

Analysis of methane emission characteristics and environmental response in natural wetlands

Moss plants (Bryophyta) are bioindicators of environmental pollution. Identification of moss plant diversity has never been carried out at Medan State University. The research design used to obtain field data is by using the exploratory method. Sampling was purposive sampling in the humid environment around Medan State University, especially in parts that have lots of trees. The results obtained in this study show that there are two types of moss plants, namely leaf moss and liverworts. There are three species of leaf moss found, namely, Sphagnum moss, Entondon seductrix, Andreaea rupestris while the liver moss found is the Bozzania trilobata species.