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Physiological and molecular diversity of feather moss associative N2-fixing cyanobacteria.

Cyanobacteria colonizing the feather moss Pleurozium schreberi were isolated from moss samples collected in northern Sweden and subjected to physiological and molecular characterization. Morphological studies of isolated and moss-associated cyanobacteria were carried out by light microscopy. Molecular tools were used for cyanobacteria identification, and a reconstitution experiment of the association between non-associative mosses and cyanobacteria was conducted. The influence of temperature on N2 fixation in the different cyanobacterial isolates and the influence of light and temperature on N2-fixation rates in the moss were studied using the acetylene reduction assay. Two different cyanobacteria were effectively isolated from P. schreberi: Nostoc sp. and Calothrix sp. A third genus, Stigonema sp. was identified by microscopy, but could not be isolated. The Nostoc sp. was found to fix N2 at lower temperatures than Calothrix sp. Nostoc sp. and Stigonema sp. were the predominant cyanobacteria colonizing the moss. The attempt to reconstitute the association between the moss and cyanobacteria was successful. The two isolated genera of cyanobacteria in feather moss samples collected in northern Sweden differ in their temperature optima, which may have important ecological implications.

Base Sequence↗

Plant desiccation and protein synthesis: an in vitro system from dry and hydrated mosses using endogenous and synthetic messenger ribonucleic Acid.

The conditions and requirements for an in vitro protein synthesizing system from the moss Tortula ruralis are outlined. Using this system the effects of desiccation, achieved quickly or slowly, were studied. Slowly dried moss retained fewer polyribosomes on desiccation but more active ribosomes than rapidly dried moss. Even in the completely desiccated moss the polyribosomes and/or free ribosomes present have retained their synthetic capacities. On rehydration, the slowly dried moss resumed protein synthesis more quickly than moss previously desiccated rapidly. Moss ribosomes are cycloheximide sensitive and chloramphenicol insensitive and thus the major protein synthesis occurs within the cytoplasm on rehydration. Extracted polyribosomes per se can withstand desiccation to a significant extent, suggesting that protection by the cytoplasm might not be necessary. The aquatic moss Hygrohypnum luridum can retain polyribosomal and ribosomal activity during desiccation, but this decreases greatly on rehydration.

Journal Article↗

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↗

Seasonal and annual distribution of organic contaminants in marine sediments from Elkhorn slough, moss landing harbor and nearshore Monterey Bay, California.

This 3-year study provides data on the spatial, seasonal and annual variability of hydrocarbons and total organic carbon present in marine sediments at three sites: Elkhorn Slough, Moss Landing Harbor and nearshore Monterey Bay in the vicinity of Moss Landing, California. The study provides baseline information that could be used to evaluate the potential impacts of future fuel oil releases occuring in the Moss Landing area. Groups of hydrocarbons were chosen to represent the hydrocarbon inputs into the Moss Landing area. These included the pesticide dichlorodiphenyltrichloroethane (DDT), polychlorinated biphenyl (PCB), phthalic acid ester (PAE), polycyclic aromatic hydrocarbon (PAH) and combustion PAHs (SigmaCOMBs). For SigmaDDTs, SigmaPCBs, SigmaPAEs, SigmaPAHs and SigmaCOMBs, the major sources of variability were between sites and random effects. Subsites within each site contributed little variability. No significant seasonal differences in any chemical contaminant group were found at any site. Significant seasonal differences in total organic carbon (TOC) and significant annual differences in SigmaPCBs, SigmaPAHs, SigmaCOMBs and SigmaPAEs were found at the nearshore Monterey Bay site. Significant annual differences in SigmaPAEs and TOC were found within Moss Landing Harbor, and significant annual differences in SigmaPAEs were found within the Elkhorn Slough site. Implications for future sampling designs in the Moss Landing area are that given the current baseline conditions (a stable, low rate of hydrocarbon input), a variability of 75-150 m(2) may not need to be heavily sampled. Spatial variability, not seasonal or annual variability, is the major source of hydrocarbon variability in Moss Landing sediments, although 3 years may not be long enough to establish long-term annual trends. Further research to determine the SigmaPAH spatial sampling scale for oil spills is needed.

Journal Article↗

Estimation of metal uptake efficiencies from precipitation in mosses in Lithuania

The main sources contributing to heavy metal content in mosses in Lithuania were examined by a comparison of heavy metal concentrations in moss and corresponding deposition levels calculated from bulk deposition analysis. Bulk deposition was collected in open areas as well as under the canopy of trees. Uptake efficiencies in moss were calculated for Cd, Cr, Cu, Fe, Mn, Ni, V and Zn. All elements in moss except Pb and Cd appeared to be more or less influenced by sources other than air pollution. The general order of this influence on the heavy metal content in moss was observed as follows: Ni < V < Cr < Zn < Fe < Mn. The contents of Mn and Zn in moss were greatly influenced by leaching from the canopy while Pb was the only element which showed a net metal retention by the canopy. Concentrations of Fe and Cr in moss were dominating due to contribution from soil dust.

Journal Article↗

Deposition and disease: a moss monitoring project as an approach to ascertaining potential connections.

In the years 1995 and 1996 the atmospheric deposition of elements in the EuroRegion Neisse (ERN) was determined in a biomonitoring project using mosses. The mosses Pleurozium schreberi and Polytrichum formosum were chosen as biomonitors because of their wide distribution in the area studied. The moss samples were analysed by ICP-MS and ICP-OES for their concentrations of 37 chemical elements. The results were shown in the form of maps. The data from the moss monitoring project served as a basis for determining those elements in the deposited material that promote the occurrence of disease. This was done by correlating the figures for the various diseases with the appropriate element concentrations in the mosses. Indications were found that a connection exists between the thallium content of mosses and the occurrence of cardiovascular disease and between Ce, Fe, Ga and Ge levels in the mosses and the incidence of diseases of the respiratory system.

Adolescent↗

Aquatic mosses as a monitoring tool for 137Cs contamination in streams and rivers-a field study from central southern Norway.

Mosses are frequently used as biomonitors for trace element pollution in the aquatic environment. The purpose of this study was to assess their usefulness as a tool in monitoring and in regional surveys of radioactive contamination. Specimens of the aquatic mosses, Fontinalis antipyretica and F. dalecarlica, were transplanted from non-contaminated areas to streams and rivers in the Norwegian Jotunheimen Mountains and neighbouring lowland areas that had received radioactive fallout after the Chernobyl accident. Equilibrium concentrations of 137Cs in the exposed mosses were reached after a few weeks. Two series from 20 streams in 1994 and 1996 show linear correlations between activities in water and moss samples and biomagnification ratios of 10(4) - 10(5). We conclude that mosses are better suited for monitoring purposes than water samples, because they provide values integrated over weeks while the radioactivity in surface waters can be subject to rapid variations according to hydrological events. The activity concentrations in aquatic mosses can be easily measured with good precision even when aqueous concentrations are below the limit of detection. Use of aquatic mosses also reduces the logistic problems of transporting large volumes of water, especially in areas inaccessible by road.

Bryopsida↗

Influence of current velocity on cadmium accumulation by an aquatic moss and the consequences for its use as a biomonitor.

Aquatic mosses are widely used as biomonitors of contaminant concentrations in running waters. The results of several previous studies suggest that metal concentrations in mosses are influenced by current velocity and that this variable should be taken into account when mosses are used as metal biomonitors. However, in these studies, the purported influence of water velocity was confounded by other uncontrolled variables. We conducted our study to test the influence of current velocity on Cd accumulation by the riverine moss Fontinalis dalecarlica. We found no difference in Cd accumulation over 2 weeks by moss exposed in the laboratory to a constant Cd concentration over a wide range of current velocities (0.01-0.70 m s-1) that occur in the field. Similarly, the results of a field experiment, in which we exposed F. dalecarlica in a Cd-contaminated creek to four current velocities (0.05-0.50 m s-1), confirmed that in nature Cd accumulation by this moss is not influenced by current velocity. We show that a bioaccumulation model that ignores current velocity describes Cd accumulation by F. dalecarlica very well. Our results suggest that current velocity does not have to be considered when using aquatic mosses as metal biomonitors.

Biological Availability↗

Modeling cadmium exchange by an aquatic moss (Fontinalis dalecarlica).

Although aquatic mosses are widely used as metal biomonitors in rivers, there are few effective models to describe metal uptake and loss by these plants. To fill this gap, we exposed the aquatic moss Fontinalis dalecarlica for 28 d to three Cd concentrations (approximately 5-50 nM) in a flow-through laboratory system. Cadmium accumulation by F. dalecarlica was rapid during the first few days of exposure and slowed thereafter but did not reach a steady state within our 1-month long experiment. This lack of a plateau in moss concentrations suggests that, for biomonitoring purposes, the duration of moss exposure should be considered either through a model of the type that we tested or by standardizing the exposure time of mosses transplanted in the field. During the subsequent 22-d elimination phase of our experiment, Cd concentrations in mosses did not return to their initial levels. This result suggests that a two-compartment model is likely to be more effective at describing Cd losses than would a one-compartment alternative. Indeed, predictions of a two-compartment model closely fitted our experimental data, which augurs well for the wider use of this model for other moss species and metals.

Bryophyta↗

Molecular evolution and phylogeny of the atpB-rbcL spacer of chloroplast DNA in the true mosses.

The nucleotide variation of a noncoding region between the atpB and rbcL genes of the chloroplast genome was used to estimate the phylogeny of 11 species of true mosses (subclass Bryidae). The A+T rich (82.6%) spacer sequence is conserved with 48% of bases showing no variation between the ingroup and outgroup. Rooted at liverworts, Marchantia and Bazzania, the monophyly of true mosses was supported cladistically and statistically. A nonparametric Wilcoxon Signed-Ranks test Ts statistic for testing the taxonomic congruence showed no significant differences between gene trees and organism trees as well as between parsimony trees and neighbor-joining trees. The reconstructed phylogeny based on the atpB-rbcL spacer sequences indicated the validity of the division of acrocarpous and pleurocarpous mosses. The size of the chloroplast spacer in mosses fits into an evolutionary trend of increasing spacer length from liverworts through ferns to seed plants. According to the relative rate tests, the hypothesis of a molecular clock was supported in all species except for Thuidium, which evolved relatively fast. The evolutionary rate of the chloroplast DNA spacer in mosses was estimated to be (1.12 +/- 0.019) x 10(-10) nucleotides per site per year, which is close to the nonsynonymous substitution rates of the rbcL gene in the vascular plants. The constrained molecular evolution (total nucleotide substitutions, K approximately 0.0248) of the chloroplast DNA spacer is consistent with the slow evolution in morphological traits of mosses. Based on the calibrated evolutionary rate, the time of the divergence of true mosses was estimated to have been as early as 220 million years ago.

AT Rich Sequence↗

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↗

Contact allergy to oak moss: search for sensitizing molecules using combined bioassay-guided chemical fractionation, GC-MS, and structure-activity relationship analysis.

In addition to pure synthetic fragrance materials several natural extracts are still in use in the perfume industry. Among them oak moss absolute, prepared from the lichen Evernia prunastri (L.) Arch., is considered a major contact sensitizer and is therefore included in the fragrance mix used for diagnosing perfume allergy. The process of preparing oak moss absolute has changed during recent years and, even though several potential sensitizers have been identified from former benzene extracts, its present constituents and their allergenic status are not clear. In the study reported here, we applied a method developed for the identification of contact allergens present in natural complex mixtures to oak moss absolute. The method is based on the combination of bioassay-guided chemical fractionation, gas chromatography-mass spectrometry analysis and structure-activity relationship studies. Our first results showed that atranol and chloroatranol, formed by transesterification and decarboxylation of the lichen depsides, atranorin and chloroatranorin, during the preparation of oak moss absolute, are strong elicitants in most patients sensitized to oak moss. Methyl-beta-orcinol carboxylate, a depside degradation product and the most important monoaryl derivative of oak moss from an olfactory standpoint, was also found to elicit a reaction in most patients.

Allergens↗

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↗

Cloning and nucleotide sequence analysis of genes coding for the major chlorophyll-binding protein of the moss Physcomitrella patens and the halotolerant alga Dunaliella salina.

Two clones have been isolated from a genomic library of the moss Physcomitrella patens and a cDNA library of the halotolerant green alga Dunaliella salina. The isolates contain genes coding for the major light-harvesting chlorophyll-a/b-binding protein (CAB) in the photosystem II (PSII) light-harvesting complex (LHCII). The 2544-bp insert of the moss genomic clone contains the complete CAB-coding region and 5' and 3' flanking sequences. The coding region contains an intron of 359 bp which is spanned by a pair of 9-bp perfect direct repeats. There are two CCAAT boxes and five enhancer-like elements related to (G)TGGTTTAAA(G) (Weiher et al., 1983) residing in the intron. Comparisons of the moss cab gene with sequences of light-inducible genes of higher plants reveal homologous and repeated sequences similar to the enhancer element in the 5' region upstream from the TATA and CCAAT boxes thought to be responsive to light inducibility. The 1256-bp algal cDNA contains the complete CAB-coding sequence, a 170-bp 5'-nontranslated region, and a 264-bp 3'-nontranslated region. While the overall homology in the nontranslated regions is low between the cab gene of the moss and that of the alga, the 3'-nontranslated regions of the two contain some sequences that are conserved among the cab genes in higher plants. The deduced amino acid sequences of these two clones are highly conserved except for the N-terminal region. Their hydropathic plots are very similar and both possess three hydrophobic segments that are likely alpha-helical transmembrane segments. The proposed CAB transit peptide sequence of the alga is divergent from that of the moss or higher plants, suggesting that they may have evolved from different origins. Southern blot analysis shows that the cab genes in the moss and the alga, as in higher plants, are encoded by a number of homologous genes constituting a multigene family.

Amino Acid Sequence↗

Reconstructing temporal trends in heavy metal deposition: assessing the value of herbarium moss samples.

The use of the herbarium moss archive for investigating past atmospheric deposition of Ni, Cu, Zn, As, Cd and Pb was evaluated. Moss samples from five UK regions collected over 150 years were analysed for 26 elements using ICP-MS. Principal components analysis identified soil as a significant source of Ni and As and atmospheric deposition as the main source of Pb and Cu. Sources of Zn and Cd concentrations were identified to be at least partly atmospheric, but require further investigation. Temporal and spatial trends in metal concentrations in herbarium mosses showed that the highest Pb and Cu levels are found in Northern England in the late 19th century. Metal concentrations in herbarium moss samples were consistently higher than those in mosses collected from the field in 2000. Herbarium moss samples are concluded to be a useful resource to contribute to reconstructing trends in Pb and Cu deposition, but not, without further analysis, for Cd, Zn, As and Ni.

Air Pollutants↗

Temporal and spatial trends in Hg deposition monitored by moss analysis.

Results for Hg from a nation-wide survey of atmospheric deposition of heavy metals in Norway in 2000 are presented and compared to similar results from 1985, 1990 and 1995. The study is based on sampling of the naturally growing feather moss Hylocomium splendens at approximately 500 sites all over the country. Cold vapor AAS (1985, 1990) or atomic fluorescence spectrometry (1995, 2000) was used for Hg determinations after HNO(3) digestion of the moss samples. As different from the wet deposition of Hg and several other heavy metals in Norway the Hg concentration in moss shows no significant north-south gradient. This is explained by a considerable supply of Hg to the moss from dry deposition of Hg(0) in addition to the retention of Hg(2+) from wet deposition. Presumably the cool summer climate particularly in the middle and northern parts of the country explains that some of the Hg(0) retained in the moss is not lost by re-volatilisation. Whereas the level and geographic distribution of Hg in moss showed only small differences during 1985-1995 the 2000 data are approximately 30% lower. A satisfactory explanation of this difference remains to be found.

Air Pollutants↗

A comparison of indigenous mosses and topsoils for use in monitoring atmospheric heavy metal deposition in Galicia (northwest Spain).

Samples of moss (Scleropodium purum or, in its absence, Hypnum cupressiforme) and of topsoil (0-5 cm) were collected in the spring of 1995, from 75 sites distributed throughout Galicia (northwest Spain) in order to compare the potential usefulness of each in evaluating the atmospheric deposition of contaminants. Analysis was made of the total content of various metals in the moss and of different soil metal fractions (including total metal content, labile metals and metals bound to organic material). We found that the labile fraction most closely resembled the metal content of the moss. Calculation of enrichment factors allowed us to show that the soil was not, except in the case of Cr, the main source of contamination in the moss. By comparing contamination factors of metals in moss and soil fraction samples we found that moss samples were more useful for evaluating the degree of contamination at different sites.

Air Pollutants↗

Distribution of trace metals in moss biomonitors and assessment of contamination sources in Portugal.

A biomonitoring survey using the moss species [Hypnum cupressiforme Hedw. and Scelopodium touretii (Brid.) L. Kock] was performed in the whole territory of Portugal, in order to evaluate the atmospheric deposition of the following elements: Cd. Cr, Cu, Fe, Mn, Ni, Pb and Zn. The concentrations of the same elements were also obtained in two types of soil samples, collected under the moss and in nearby plots without any plant coverage, and relationships between moss and soil concentrations was investigated using the multivariate statistical method of Co-inertia Analysis. Also, relationships between concentrations in moss and several anthropogenic, geologic, pedologic and environmental parameters were screened using the same method of Co-inertia Analysis. Higher concentrations of Cu, Pb and Zn were found in areas of higher population density, with higher gasoline consumption, while higher values of Fe and Cr occur in the driest region, with lower plant coverage, indicating strong contamination by resuspended soil particles. Results also show good agreement between moss and soil contents, even for elements with high contribution of anthropogenic sources. The spatial pattern in Portugal of element contents in mosses were also detected and discussed in relation to local contamination sources.

Absorption↗