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A procedure to purify and culture a clonal strain of the aquatic moss Fontinalis antipyretica for use as a bioindicator of heavy metals.

A procedure to culture the aquatic moss, Fontinalis antipyretica, is described. The moss was collected in a stream, carried to the laboratory, and immediately treated: apices were isolated and rinsed under a stereomicroscope and submitted to ultrasonic cleaning combined with an iodine treatment and then to an antibiotic/anti-eukaryotic treatment for 4 days. This procedure permitted us for the first time to eliminate the microalgae associated with an aquatic moss in culture, solving the problem of proliferation of these microalgae in the culture medium. Furthermore, the minimal concentration of eight essential metals required by this moss was determined in order to have metal concentrations in the cultured moss as low as possible. A perennial culture of a clonal strain of an aquatic moss for use as a metal bioaccumulator was obtained and the first step to provide a standardized bioindication tool for metal detection in continental waters was passed over.

Biological Assay↗

Nitrogen fixation and leaching of biological soil crust communities in mesic temperate soils.

Biological soil crust is composed of lichens, cyanobacteria, green algae, mosses, and fungi. Although crusts are a dominant source of nitrogen (N) in arid ecosystems, this study is among the first to demonstrate their contribution to N availability in xeric temperate habitats. The study site is located in Lucas County of Northwest Ohio. Using an acetylene reduction technique, we demonstrated potential N fixation for these crusts covering sandy, acidic, low N soil. Similar fixation rates were observed for crust whether dominated by moss, lichen, or bare soil. N inputs from biological crusts in northwestern Ohio are comparable to those in arid regions, but contribute substantially less N than by atmospheric deposition. Nitrate and ammonium leaching from the crust layer were quantified using ion exchange resin bags inserted within intact soil cores at 4 cm depth. Leaching of ammonium was greater and nitrate less in lichen than moss crusts or bare soil, and was less than that deposited from atmospheric sources. Therefore, biological crusts in these mesic, temperate soils may be immobilizing excess ammonium and nitrate that would otherwise be leached through the sandy soil. Moreover, automated monitoring of microclimate in the surface 7 cm of soil suggests that moisture and temperature fluctuations in soil are moderated under crust compared to bare soil without crust. We conclude that biological crusts in northwestern Ohio contribute potential N fixation, reduce N leaching, and moderate soil microclimate.

Biodegradation, Environmental↗

Isolation and characterisation of three moss-derived beta-tubulin promoters suitable for recombinant expression.

The moss Physcomitrella patens is an excellent tool to study plant gene-function relationships due to its high rate of homologous recombination (HR). It has also been shown to be very useful in the production of recombinant proteins which are secreted into a simple medium. Thus, there is a need for suitable promoters functional in this well established model organism. We isolated genomic flanking regions of the beta-tubulin gene family from Physcomitrella, concentrating on those family members showing high transcript abundance integrated over gametophytic tissues. Using a novel, fast and reliable quantification assay based on the transient expression and secretion of a recombinant human protein, three genomic upstream regions were characterised in serial deletion constructs. Expression rates were up to three times higher than those obtained with the 35S cauliflower mosaic virus (35S) promoter, which served as a reference.

5' Untranslated Regions↗

From axenic spore germination to molecular farming. One century of bryophyte in vitro culture.

The first bryophyte tissue culture techniques were established almost a century ago. All of the techniques that have been developed for tissue culture of seed plants have also been adapted for bryophytes, and these range from mere axenic culture to molecular farming. However, specific characteristics of bryophyte biology--for example, a unique regeneration capacity--have also resulted in the development of methodologies and techniques different than those used for seed plants. In this review we provide an overview of the application of in vitro techniques to bryophytes, emphasising the differences as well as the similarities between bryophytes and seed plants. These are discussed within the framework of physiological and developmental processes as well as with respect to potential applications in plant biotechnology.

Bryophyta↗

Decrease in excitability of LG following habituation of the crayfish escape reaction.

Crayfish escapes from threatening stimuli to the abdomen by tailflipping upwards and forwards. This lateral giant (LG)-mediated escape reaction habituates readily upon repetitive sensory stimulation. Using an isolated abdominal nerve cord preparation, we have analyzed the change in LG activity by applying additional sensory stimulation after different periods following habituation to characterize the retention of LG habituation. Results show that the LG mediated response habituates more quickly, but the retention time is shorter, as repetitive sensory stimulation is applied at progressively shorter inter-stimulus time intervals. The spike response of LG recovers quickly, within several minutes after habituation, but they fail to spike when an additional stimulus is applied after specific long periods following habituation. The critical period of the delay for this decrease in excitability of LG is dependent on the inter-stimulus time interval of the initial repetitive stimulus. As the inter-stimulus interval became longer, the delay needed for decrease in excitability became shorter. Furthermore, the local injection of 10(-6) mol l-1 octopamine into the neuropil just following habituation promotes the achievement of decrease in excitability. No effects were observed when 10(-6) mol l-1 serotonin and tyramine were injected. These results suggested octopamine promotes decrease in excitability of LG following habituation.

Animals↗

Moisture conditions and the presence of bryophytes determine fescue species abundance in a dry calcareous grassland.

Festuca ovina is the abundant matrix-forming species and F. rubra a subordinate species in shallow-soil calcareous grasslands. F. pratensis is a transient species, occurring sparsely in this community. We hypothesised that the different abundances of these three species are primarily due to the differential effect of moisture conditions on their germination and early establishment, and that the effect of the pattern of rainfall intensity depends on the presence or absence of a bryophyte layer. We studied the dependence of the germination and establishment of the three fescue species on the moisture conditions both in the laboratory and in the patches of intact grassland community (microcosms). In a laboratory germination experiment, F. pratensis showed the highest, F. rubra, the intermediate and F. ovina, the lowest drought tolerance. In microcosms, the establishment of F. ovina was the highest. At the same time, the annual mortality of seedlings of F. ovina was the lowest. All three species responded positively to an increasing irrigation level. Differently from F. ovina, F. rubra showed a positive response only in plots from which the bryophyte layer had been removed, while F. pratensis responded positively to both irrigation and bryophyte removal. We conclude that moisture conditions have a differential effect on the three fescue species mainly in the seedling establishment, not in the germination phase. For the successful establishment of F. rubra and F. pratensis, the coincidence of high rainfall and local disturbance, removing bryophytes, is required. The presence or absence of bryophytes had no effect on establishment in dry years, while in rainy years the removal of bryophytes has a clear positive effect.

Bryophyta↗

Habitat modification by the stream macrophyte Justicia americana and its effects on biota.

We conducted an in-stream experiment to determine if and by what mechanisms the emergent macrophyte, Justicia americiana, enhances streambed stability and influences associated benthic organisms. Treatments included removal of aboveground stems, removal of stems and rhizomes, and a control. Stone stability and embeddedness were higher within intact patches of Justicia compared with areas where stems and rhizomes were removed. Presence of belowground structures binding stones and higher embeddedness were responsible for differences in stone stability among treatments. Sediment deposition was highly variable among treatments; however, silt and clay deposition was highest in the control when Justicia stems reached maximum biomass. Response of neighboring benthic organisms to Justicia treatments varied with organism mobility and trophic level. Sedentary unionid mussels were more abundant in the control than in the stem and rhizome-removal treatment, whereas abundance of mobile snails (Elimia ucheensis) and sessile plants (Podostemon ceratophyllum and mosses) did not differ among treatments. Justicia appears to modify the stream environment by: (1) increasing stability of streambed sediments, and (2) reducing current velocity, thereby enhancing deposition of fine sediments and organic matter.

Acanthaceae↗

Mosses and the struggle for light in a nitrogen-polluted world.

The impact of reduced light conditions as an indirect effect of nitrogen (N) deposition was determined on three mosses in a montane ecosystem, where sedge and grass cover increase due to N enrichment. Additionally, in the greenhouse we established the importance of low light to moss growth as an indirect N deposition effect relative to the direct toxic effects of N. The amount of light reaching the moss layer was strongly and negatively related to graminoid abundance. Mosses showed differing sensitivities to reduced light in the field. Racomitrium lanuginosum biomass was found to be highest under high-light conditions, Polytrichum alpinum at intermediate light levels, whilst that of Dicranum fuscescens was unrelated to light availability. Moreover, Racomitrium biomass decreased with increasing amounts of graminoid litter, whereas the other species were little affected. All three mosses responded differently to the combination of elevated N (20 vs 10 kg N ha(-1) year(-1)) and reduced light (60 and 80% reduction) in the greenhouse. Racomitrium growth was strongly influenced by both light reduction and elevated N, in combination reducing shoot biomass up to 76%. There was a tendency for Dicranum growth to be modestly reduced by elevated N when shaded, causing up to 19% growth reduction. Polytrichum growth was not influenced by elevated N but was reduced up to 40% by shading. We conclude that competition for light, induced by vascular plants, can strongly influence moss performance even in unproductive low biomass ecosystems. The effects of reduced light arising from N pollution can be as important to mosses as direct toxicity from N deposition. Yet, different sensitivities of mosses to both toxic and shading effects of elevated N prevent generalisation and can lead to competitive species replacement within moss communities. This study demonstrates the importance of understanding moss-vascular plant interactions to allow interpretation and prediction of ecosystem responses to anthropogenic drivers such as atmospheric N deposition or climate change.

Bryophyta↗

Differential effects of lichens, mosses and grasses on respiration and nitrogen mineralization in soils of the New Jersey Pinelands.

In the New Jersey Pinelands, severely disturbed areas often do not undergo a rapid succession to forest; rather, a patchy cover of lichens, mosses and grasses persists for decades. We hypothesized that these plant covers affect soil microbial processes in different ways, and that these effects may alter the successional dynamics of the patches. We predicted that the moss and grass covers stimulate soil microbial activity, whereas lichens inhibit it, which may in turn inhibit succession. We collected soil cores from beneath each type of cover plus bare soil within two types of highly disturbed areas--sites subjected to hot wildfires, and areas mined for sand. Organic matter (OM) content, soil respiration and potential N mineralization were measured in the cores. Soils under mosses were similar to those under grasses; they accumulated more OM and produced more mineral N, predominantly in the form of ammonium, than either the bare soils or the soils beneath lichens. Mineralization under lichens, like that of the bare soils but unlike the soils beneath mosses or grasses, was dominated by net nitrification. These patterns were reproduced in experimentally transplanted moss and lichen mats. Mosses appear to create high-nutrient microsites via high rates of OM accumulation and production of ammonium, whereas lichens maintain low-nutrient patches similar to bare soil via low OM accumulation rates and production of mineral N predominantly in the mobile nitrate form. These differences in soil properties may explain the lack of vascular plant invasion in lichen mats, in contrast to the moss-dominated areas.

Analysis of Variance↗

Improved water retention links high species richness with increased productivity in arctic tundra moss communities.

A positive relationship between plant species richness and ecosystem functioning has been found in a number of experimental studies. Positive species interactions at high species numbers have been suggested as a cause, but mechanisms driving positive interactions have not often been tested. In this experiment we asked three questions: (1) What is the relationship between species richness and productivity in experimentally constructed moss communities? (2) Is this relationship affected by plant density? and (3) Can changes in moisture absorption and retention explain observed relationships? To answer these questions we exposed arctic tundra moss communities of different species richness levels (1-11 species) and two different densities in the greenhouse to two levels of drought (short and long). Biomass (by the community and individual species), height and community moisture absorption and retention were measured as response variables. High species diversity increased productivity (more so in low-density plots than in high-density plots), but only when plots were watered regularly. Plot moisture retention was improved at high species richness as well, and plant height and variation in height was increased compared to plants in monoculture. Under high-density and short-drought conditions 10 out of 12 species grew better in mixture than in monoculture, but under the long drought treatment only six species did. A positive feedback loop between biomass and improved humidity under high diversity was supported by path analysis. We conclude that in this community the relationship between species richness and productivity depends on moisture availability and density, with improved water absorption and retention likely to be the mechanism for increased plant growth when drought periods are short. Furthermore, since this is the opposite of what has been found for temperate moss communities, conclusions from one system cannot automatically be extrapolated to other systems.

Arctic Regions↗

Factors influencing reproductive success in the clonal moss, Hylocomium splendens.

Female reproductive success in the unisexual perennial clonal moss Hylocomium splendens was examined by recording, if the segment was reproductive [produced sporophyte(s)] or not, together with several distance-to-male and male density variables, and segment size. This was done for every female segment in a population over a 5 year study period. A high fraction of the population could be sexed because we monitored the population in situ for 5 years, and thereafter harvested the population for electrophoretic analysis from which the clonal identity and expressed sex could be deduced. Fertilization distances in H. splendens were short, indicated by the fact that as many as 85% of the female segments with sporophytes were situated within a distance of 5.0 cm from the nearest male. The longest distance measured between a sporophytic female and the closest male was 11.6 cm. However, analysed within a generalized linear modelling (GLM) framework, the year was the best single predictor for the presence of H. splendens sporophyte although female-segment size and distance to the closest situated male were also strongly significant. The two latter factors explained larger fractions of variation in sporophyte presence in a GLM model with three predictors than in single-predictor models. This is because (i) the large variation in sporophyte production among years partly obscures the strong general increase in sporophyte production with increasing female-segment size and vitality, and (ii) the between-year variation and the size obscure the effect of the distance to the most proximate male. To our knowledge, this study is the first to incorporate into one model the relative importance of several factors for bryophyte reproductive success. Our results demonstrate the value of multiple-predictor approaches in studies of reproductive success.

Bryophyta↗

Estimation of stand-level leaf area for boreal bryophytes.

Bryophytes dominate the carbon and nitrogen cycling of many poorly drained terrestrial ecosystems and are important in the vegetation-atmosphere exchange of carbon and water, yet few studies have estimated their leaf area at the stand scale. This study quantified the bryophyte-specific leaf area (SLA) and leaf area index (LAI) in a group of different-aged boreal forest stands in well and poorly drained soils. Species-specific SLA (for three feather mosses, four Sphagnum spp. and Aulacomnium palustre mixed with Tomentypnum nitens) was assessed by determining the projected area using a flatbed scanner and cross-sectional geometry using a dissecting microscope. The hemisurface leaf area was computed as the product of SLA and live biomass and was scaled by coverage data collected at all stands. Pleurozium schreberi dominated the spatial coverage, biomass and leaf area in the well-drained stands, particularly the oldest, while S. fuscum and A. palustre were important in the poorly drained stands. Live moss biomass ranged from 47 to 230 g m(-2) in the well-drained stands dominated by feather mosses and from 102 to 228 g m(-2) in the poorly drained stands. Bryophyte SLA varied between 135 and 473 cm(2) g(-1), for A. palustre and S. capillifolium, respectively. SLA was strongly and significantly affected by bryophyte species, but did not vary between stands; in general, there was no significant difference between the SLA of non-Sphagnum mosses. Bryophyte LAI increased with stand age, peaking at 3.1 and 3.7 in the well and poorly drained stands, respectively; this represented approximately 40% of the overstory LAI in the well-drained stands and 100-1,000% in the poorly drained stands, underscoring the important role bryophytes play in the water and carbon budgets of these boreal forests.

Bryophyta↗

Bryophyte dispersal by flying foxes: a novel discovery.

This research provides the first evidence of dispersal of bryophytes and associated microorganisms through ingestion by a highly mobile vertebrate vector, the spectacled flying fox (Pteropus conspicillatus). Bryophyte fragments were found in faeces collected at four P. conspicillatus' camps in the Wet Tropics bioregion, northeastern Australia. These fragments were viable when grown in culture; live invertebrates and other organisms were also present. Our study has significantly increased understanding of the role of flying foxes as dispersal vectors in tropical forests.

Animals↗

A new approach to assess atmospheric nitrogen deposition by way of standardized exposition of mosses.

A standardized method of moss exposition for assessing variations in nitrogen deposition was tested in the western part of Germany. Six pleurocarpous moss species were transplanted to two sites differing in their deposition rates but being comparable as to their climatic conditions. The mosses were exposed in standardized containers over a period of 12 months; the focus of interest was the effect of N deposition on nitrogen content and on (15)N natural abundance (delta(15)N-values). Within the first nine months only trends could be observed. However, after one year all species tested showed significantly higher N concentrations at the highly polluted site. Besides, more negative delta(15)N-values possibly reflecting the higher ammonium input were detected at this site. Surprisingly, most of the plants though being kept in plastic containers without a favourable substratum did not show any conspicuous deficiency symptoms. The potential advantages of a standardized moss exposition for N monitoring purposes are discussed. It is concluded that the method presented here can yield significant results in particular if the number of testing sites is increased.

Atmosphere↗

Assessment of atmospheric deposition of heavy metals and other elements in Belgrade using the moss biomonitoring technique and neutron activation analysis.

This study aimed at assessing atmospheric deposition of heavy metals and other elements using the moss genera Brachythecium sp. (B. rutabulum and B. salebrosum) and Eurhynchium sp. (E. hians and E. striatum) collected in autumn 2004 in the urban area of Belgrade. The concentrations of 36 elements (Na, Mg, Al, Cl, K, Ca, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Zn, As, Se, Br, Rb, Sr, Mo, Sb, I, Cs, Ba, La, Ce, Sm, Tb, Dy, Hf, Ta, W, Hg, Th, U) were determined in moss and local topsoil samples by instrumental neutron activation analysis. The concentration of elements in moss positively correlated to those obtained for topsoil. High enrichment factors for As, Zn, Mo, Br, Sb, Se, Hg and Cl, calculated to continental crust composition, gave an evidence for anthropogenic impact on urban area, mainly due to intensive vehicular traffic and fossil fuel combustion. The concentration of elements in moss, characteristic for fossil fuel combustion, obtained in this study were substantially lower than in the previous investigation (2000) conducted in the area of Belgrade. The level of concentrations for V, Cr, Ni, and As in moss from this study correlated to those measured for neighboring countries, and were several times higher than the base-level data from low polluted areas. The level of accumulated elements in both investigated moss genera were similar and all studied species could be combined for biomonitoring purposes in urban areas.

Air Pollutants↗

Implementation of neural networks for classification of moss and lichen samples on the basis of gamma-ray spectrometric analysis.

Mosses and lichens have an important role in biomonitoring. The objective of this study is to develop a neural network model to classify these plants according to geographical origin. A three-layer feed-forward neural network was used. The activities of radionuclides ((226)Ra, (238)U, (235)U, (40)K, (232)Th, (134)Cs, (137)Cs and (7)Be) detected in plant samples by gamma-ray spectrometry were used as inputs for neural network. Five different training algorithms with different number of samples in training sets were tested and compared, in order to find the one with the minimum root mean square error. The best predictive power for the classification of plants from 12 regions was achieved using a network with 5 hidden layer nodes and 3,000 training epochs, using the online back-propagation randomized training algorithm. Implementation of this model to experimental data resulted in satisfactory classification of moss and lichen samples in terms of their geographical origin. The average classification rate obtained in this study was (90.7 +/- 4.8)%.

Bryophyta↗

The cellulose synthase (CESA) gene superfamily of the moss Physcomitrella patens.

The CESA gene superfamily of Arabidopsis and other seed plants comprises the CESA family, which encodes the catalytic subunits of cellulose synthase, and eight families of CESA-like (CSL) genes whose functions are largely unknown. The CSL genes have been proposed to encode processive beta-glycosyl transferases that synthesize noncellulosic cell wall polysaccharides. BLAST searches of EST and shotgun genomic sequences from the moss Physcomitrella patens (Hedw.) B.S.G. were used to identify genes with high similarity to vascular plant CESAs, CSLAs, CSLCs, and CSLDs. However, searches using Arabidopsis CSLBs, CSLEs, and CSLGs or rice CSLFs or CSLHs as queries identified no additional CESA superfamily members in P. patens, indicating that this moss lacks representatives of these families. Intron insertion sites are highly conserved between Arabidopsis and P. patens in all four shared gene families. However, phylogenetic analysis strongly supports independent diversification of the shared families in mosses and vascular plants. The lack of orthologs of vascular plant CESAs in the P. patens genome indicates that the divergence of mosses and vascular plants predated divergence and specialization of CESAs for primary and secondary cell wall syntheses and for distinct roles within the rosette terminal complexes. In contrast to Arabidopsis, the CSLD family is highly represented among P. patens ESTs. This is consistent with the proposed function of CSLDs in tip growth and the central role of tip growth in the development of the moss protonema.

Bryophyta↗

Photochemical reactions of chlorophyll in dehydrated photosystem II: two chlorophyll forms (680 and 700 nm).

Lichens and phototolerant poikilohydric mosses differ from spinach leaves, fern fronds or photosensitive mosses in that they show strongly decreased Fo chlorophyll fluorescence after drying. This desiccation-induced fluorescence loss is rapidly reversible under rehydration. Fluorescence emission from Photosystem II at 685 nm was decreased more strongly by dehydration than 720 nm emission. Reaction centers of Photosystem II lose activity on dehydration and regain it on hydration. Heating of desiccated lichens increased Fo chlorophyll fluorescence. The activation energy for the reversible part of the temperature-dependent fluorescence increase was 0.045 eV, which corresponds to the energy difference between the 680 and 697 nm absorption bands. In desiccated chlorolichens such as Parmelia sulcata, heating induces the appearance of positive variable fluorescence related to the reversible reduction of QA due to overcoming the energy barrier. This is interpreted to provide information on the mechanism of photoprotection: energy is dissipated by changing Chl680 or P680 into a chlorophyll form, which absorbs at 700 nm and emits light at 720 nm (Chl-720 or P680(700)) with a low quantum yield. Dissipation of light energy in this trap is activated by desiccation.

Bryophyta↗