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Airborne pollutants along a roadside: assessment using snow analyses and moss bags.

Vertical snow sampling and moss bag transplants were used to estimate the local inorganic and organic pollutant load deposited from traffic along a major highway in Finland. The pH and concentrations of Cl(-), NO(3)(-), SO(4)(2-), Ca(2+), Na(+) and polyaromatic hydrocarbons (PAHs) were determined from snow samples collected in winter at different sites along the highway. In summer, moss bags containing 20 g of fresh red-stemmed feather moss (Pleurozium schreberi) were transplanted at the same sites. The moss bag transplants remained exposed to roadside traffic for a period of one month following which the samples were collected and the PAH profiles and concentrations were analysed. The deposition of inorganic and organic pollutants from road traffic was observed up to 60 m from the road. The prevailing winds had a significant effect on the dispersion of pollutants. Snow appears to be a good collector of inorganic pollutants from the atmosphere and can be used to monitor local airborne pollution from road traffic. Snow packs can also be used as passive collectors of organic pollutant loads from road traffic on a local scale. To monitor organic PAH deposition from the road traffic, moss bags appeared to be better indicators compared to snow sampling. The efficiency of moss bags in accumulating PAH compounds indicate that vegetation may be an important sink for traffic pollution.

Journal Article↗

Monitoring atmospheric metal pollution: a review of the use of mosses as indicators.

This article reviews the literature on the use of mosses for monitoring atmospheric metal pollution. It discusses the nature of mosses, the mechanisms for moss uptake of metals from the air and wet precipitation, the various forms in which mosses are used for this purpose, and cases in which mosses have been used for monitoring local hot spots of pollution, and regional patterns of metal pollution. Also highlighted are the questions of uptake of metals from the substrates, interspecies differences, and a comparison of the effectiveness of mosses with other indicator materials, among other issues.

Air Pollutants↗

Spatial analysis and indicator building for metal accumulation in mosses.

Mosses are used as passive accumulation monitors for metal accumulation in terrestrial ecosystems. Under leadership of the Federal Agency for Environmental Protection Germany took part in the previous European-wide moss monitoring campaigns 1990, 1995 and 2000. The investigations accomplished thereby cannot be presented completely in this article. The remarks rather concentrate on methodical aspects of the statistical data analysis. In Chapter 2 the design of data collection is summarized. Chapter 3 treats the geostatistical analysis and transformation of point data to areal information. Chapter 4 describes the aggregation of the element-specific metal concentrations in mosses to a spatially and temporally differentiated indicator of metal accumulation by means of descriptive and multivariate statistics. The work presented is only a small part of geostatistics and multivariate statistics which fit for analysis of moss monitoring data. Taking the results presented here as a basis, the following steps would further be of great importance: cluster-analytic evaluation of the results of the Moss Monitoring 1990 and 1995, detailing the cluster results using additional empirical and location describing information (e.g. moss species, ecoregions, site and species specific variability of metal accumulation) as well as optimizing the indicator buildung by testing of multivariate statistical regression models (e.g. Classification and Regression Trees).

Bryophyta↗

Analysis of mosses and soils for quantifying heavy metal concentrations in Sicily: a multivariate and spatial analytical approach.

BACKGROUND: The use of vegetal organisms as indicators of contamination of the environment is partially replacing traditional monitoring techniques. Amongst the vegetal organisms available, mosses appear to be good bioindicators and are used for monitoring anthropogenic and natural fall-out on soils. This study has two objectives: the evaluation of the concentrations of heavy metals in soils and mosses of the Sicily Region, in Italy and the identification of the origin of fall-out of heavy metals. METHODS: Mosses and the surface soil were sampled at 28 sites, only the youngest segments of Hylocomium splendens and Hypnum cupressiforme, corresponding to the plant tissues produced during the last 3 years, were taken. The elements Cd, Cu, Ni, Pb and Zn were analysed by ICP-MS and Hg by AAS. Statistical analysis was by PCA and spatial representation by GIS. RESULTS AND DISCUSSION: In the mosses sampled in Sicily, the highest concentrations of Cd were found around the cities of Palermo and Messina. The highest concentrations of Hg were recorded in the northern part of the island between Trapani and Messina, similar to the distribution of Cu. Different areas with the highest concentrations of Ni were found near the south coast, in the vicinity of Palermo and around the Volcano Etna. The highest concentrations of Pb were found in the south-west coast near Agrigento, where important chemical plants and petroleum refineries are located. Except for a few locations, Zn fall-out was found to be evenly distributed throughout Sicily. CONCLUSION: The sites where the concentrations of heavy metals cause greatest concern have been revealed by the PCA analysis and portrayed using GIS. Also of some concern is the diffuse and anthropogenic origin of Hg and Cd. The combined approach of using soil and mosses, together with pedological interpretation and application of multivariate statistical techniques has provided valuable insight into the environmental aspects of heavy metal deposition in a region of southern Europe. RECOMMENDATIONS AND OUTLOOK: Further insight into the deposition of heavy metals will require more detailed sampling of soils and mosses in both new and previous study areas. This needs to be complemented by detailed pedological investigations in the study areas. Future research programmes will address these issues.

Air Pollution↗

Thermal energy dissipation in reaction centres and in the antenna of photosystem II protects desiccated poikilohydric mosses against photo-oxidation.

Seasonal differences have been observed in the ability of desiccated mosses to dissipate absorbed light energy harmlessly into heat. During the dry summer season desiccation-tolerant mosses were more protected against photo-oxidative damage in the dry state than during the more humid winter season. Investigation of the differences revealed that phototolerance could be acquired or lost even under laboratory conditions. When a desiccated poikilohydric moss such as Rhytidiadelphus squarrosus is in the photosensitive state, the primary quinone, Q(A), in the reaction centre of photosystem II is readily reduced even by low intensity illumination as indicated by reversibly increased chlorophyll fluorescence. No such reduction is observed even under strong illumination in desiccated mosses after phototolerance has been acquired. In this state, reductive charge stabilization is replaced by energy dissipation. As a consequence, chlorophyll fluorescence is quenched. Different mechanisms are responsible for quenching. One is based on the presence of zeaxanthin provided drying occurs in the light. This mechanism is known to be controlled by a protonation reaction which is based on proton-coupled electron transport while the moss is still hydrated. Another mechanism which also requires light for activation, but no protonation, is activated during desiccation. While water is slowly lost, fluorescence is quenched. In this situation, an absorption band formed at 800 nm in the light is stabilized. It loses reversibility on darkening. Comparable kinetics of fluorescence quenching and 800 nm signals as well as the linear relationship between non-photochemical fluorescence quenching (NPQ) and loss of stable charge separation in photosystem II reaction centres suggested that desiccation-induced quenching is a property of photosystem II reaction centres. During desiccation, quenchers accumulate which are stable in the absence of water but revert to non-quenching molecular species on hydration. Together with zeaxanthin-dependent energy dissipation, desiccation-induced thermal energy dissipation protects desiccated poikilohydric mosses against photo-oxidation, ensuring survival during drought periods.

Bryopsida↗

Complete chloroplast DNA sequence of the moss Physcomitrella patens: evidence for the loss and relocation of rpoA from the chloroplast to the nucleus.

The complete chloroplast DNA sequence (122 890 bp) of the moss Physcomitrella patens has been determined. The genome contains 83 protein, 31 tRNA and four rRNA genes, and a pseudogene. Four protein genes (rpoA, cysA, cysT and ccsA) found in the liverwort Marchantia polymorpha and the hornwort Anthoceros formosae are absent from P.patens. The overall structure of P.patens chloroplast DNA (cpDNA) differs substantially from that of liverwort and hornwort. Compared with its close relatives, a 71 kb region from petD to rpoB of P.patens is inverted. To investigate whether this large inversion and the loss of rpoA usually occur in moss plants, we analyzed amplified cpDNA fragments from four moss species. Our data indicate that the large inversion occurs only in P.patens, whereas the loss of the rpoA gene occurs in all mosses. Moreover, we have isolated and characterized the nuclear rpoA gene encoding the alpha subunit of RNA polymerase (RNAP) from P.patens and examined its subcellular localization. When fused to green fluorescent protein, RpoA was observed in the chloroplasts of live moss protonemata cells. This indicates that chloroplast RNAP is encoded separately by chloroplast and nuclear genomes in the moss. These data provide new insights into the regulation and evolution of chloroplast transcription.

Amino Acid Sequence↗

Respiration in Relation to Adenosine Triphosphate Content during Desiccation and Rehydration of a Desiccation-tolerant and a Desiccation-intolerant Moss.

O(2) consumption by the desiccation-tolerant moss Tortula ruralis and the desiccation-intolerant Cratoneuron filicinum increased markedly during the latter stages of desiccation. ATP content of the mosses during desiccation was not correlated with O(2) consumption, but was influenced by the rate at which the mosses lost water. The more rapid the water loss, the more ATP that was present in the dry mosses. The pattern of O(2) consumption on rehydration also was influenced by the previous rate of desiccation. After rapid desiccation of T. ruralis O(2) consumption upon rehydration was considerably elevated, and for up to 24 hours. After very slow desiccation the elevation was small and brief. Normal O(2) consumption did not occur in C. filicinum after rapid desiccation, but did so within a few hours of rehydration after slower speeds of drying. ATP levels in T. ruralis returned to normal within 5 to 10 minutes of rehydration. In C. filicinum, increases in ATP were closely correlated with O(2) consumption. These observations are considered to be related to differential damage caused to mitochondria and to cellular integrity by different speeds of water loss. The desiccation-tolerant moss appears to be able to repair the severe damage imposed by rapid desiccation whereas the desiccation-intolerant moss cannot.

Journal Article↗

The density and diversity of gymnamoebae associated with terrestrial moss communities (Bryophyta: Bryopsida) in a northeastern U.S. forest.

Moss communities are commonly found in temperate forests and form a nearly continuous understory in some high latitude forests. However, little is known about the microbial component of these communities, especially the non-testate amoeboid protists. Fifty morphospecies of naked amoebae were identified in samples collected at eight sites in a northeastern North American forest. The mean number (+/-SE) of morphospecies found per sample site based on laboratory cultures was 17+/-2.1. The density of amoebae expressed as number/g dry weight of moss ranged from 3.5+/-0.04 x 10(3) to 4.3+/-0.2 x 10(4) and was positively correlated with the moss moisture content (r=0.9, P<0.001, df=26). Densities of gymnamoebae in the moss are generally higher than found in the surrounding soil, but this may be due in part to the greater weight of soil per unit volume compared with moss. The percentage of encysted forms was inversely related to the moisture content of the moss sample.

Amoebida↗

Protein encoding genes in an ancient plant: analysis of codon usage, retained genes and splice sites in a moss, Physcomitrella patens.

BACKGROUND: The moss Physcomitrella patens is an emerging plant model system due to its high rate of homologous recombination, haploidy, simple body plan, physiological properties as well as phylogenetic position. Available EST data was clustered and assembled, and provided the basis for a genome-wide analysis of protein encoding genes. RESULTS: We have clustered and assembled Physcomitrella patens EST and CDS data in order to represent the transcriptome of this non-seed plant. Clustering of the publicly available data and subsequent prediction resulted in a total of 19,081 non-redundant ORF. Of these putative transcripts, approximately 30% have a homolog in both rice and Arabidopsis transcriptome. More than 130 transcripts are not present in seed plants but can be found in other kingdoms. These potential "retained genes" might have been lost during seed plant evolution. Functional annotation of these genes reveals unequal distribution among taxonomic groups and intriguing putative functions such as cytotoxicity and nucleic acid repair. Whereas introns in the moss are larger on average than in the seed plant Arabidopsis thaliana, position and amount of introns are approximately the same. Contrary to Arabidopsis, where CDS contain on average 44% G/C, in Physcomitrella the average G/C content is 50%. Interestingly, moss orthologs of Arabidopsis genes show a significant drift of codon fraction usage, towards the seed plant. While averaged codon bias is the same in Physcomitrella and Arabidopsis, the distribution pattern is different, with 15% of moss genes being unbiased. Species-specific, sensitive and selective splice site prediction for Physcomitrella has been developed using a dataset of 368 donor and acceptor sites, utilizing a support vector machine. The prediction accuracy is better than those achieved with tools trained on Arabidopsis data. CONCLUSION: Analysis of the moss transcriptome displays differences in gene structure, codon and splice site usage in comparison with the seed plant Arabidopsis. Putative retained genes exhibit possible functions that might explain the peculiar physiological properties of mosses. Both the transcriptome representation (including a BLAST and retrieval service) and splice site prediction have been made available on http://www.cosmoss.org, setting the basis for assembly and annotation of the Physcomitrella genome, of which draft shotgun sequences will become available in 2005.

Alternative Splicing↗

Use of endogenous signal sequences for transient production and efficient secretion by moss (Physcomitrella patens) cells.

BACKGROUND: Efficient targeting to appropriate cell organelles is one of the bottlenecks for the production of recombinant proteins in plant systems. A common practice is to use the native secretory signal peptide of the heterologous protein to be produced. Though general features of secretion signals are conserved between plants and animals, the broad sequence variability among signal peptides suggests differing efficiency of signal peptide recognition. RESULTS: Aiming to improve secretion in moss bioreactors, we quantitatively compared the efficiency of two human signal peptides and six signals from recently isolated moss (Physcomitrella patens) proteins. We therefore used fusions of the different signals to heterologous reporter sequences for transient transfection of moss cells and measured the extra- and intracellular accumulation of the recombinant proteins rhVEGF and GST, respectively. Our data demonstrates an up to fivefold higher secretion efficiency with endogenous moss signals compared to the two utilised human signal peptides. CONCLUSION: From the distribution of extra- and intracellular recombinant proteins, we suggest translational inhibition during the signal recognition particle-cycle (SRP-cycle) as the most probable of several possible explanations for the decreased extracellular accumulation with the human signals. In this work, we report on the supremacy of moss secretion signals over the utilised heterologous ones within the moss-bioreactor system. Though the molecular details of this effect remain to be elucidated, our results will contribute to the improvement of molecular farming systems.

Amino Acid Sequence↗

[Distribution patterns of moss species in Shanghai city].

The mosses on the floor of 22 sites at 20 main parks and 2 chemical plants in Shanghai city were investigated and sampled. Based on 75 recorded moss species and their coverage, Two-way Indicator Species Analysis (TWINSPAN) and Detrended Correspondence Analysis (DCA) were used to analyze their distribution patterns. The results showed that the study sites could be identified as three groups. Group 1 included two chemical plants and two parks at the center of the city, with 23 moss species and the smallest total coverage of 21.29%; group 2 included part of the parks at the center of the city and surrounding areas, with 44 moss species and total coverage of 37.94%; and group 3 mainly included the parks at surrounding areas and suburbs of the city, with 49 moss species and total coverage of 49.66%. The results reflected a certain correlation of the distribution of moss species with different habitats and polluted and disturbed environments in the city.

Bryophyta↗

Cutaneous sporotrichosis in forestry workers. Epidemic due to contaminated Sphagnum moss.

In December 1975 and January and February 1976, an epidemic of cutaneous sporotrichosis occurred in Mississippi among forestry workers and other persons exposed to sphagnum moss used in packing pine seedlings. Seventeen cases were identified, 15 of which were from patients who had been exposed to sphagnum moss from a single source. Attack rates were significantly higher among workers exposed to this moss than among those not exposed. Sporothrix schenckii was cultured from the implicated batch of sphagnum moss but not from other batches. The source of contamination of the sphagnum moss that caused this epidemic and sphagnum moss associated with similar epidemics is unknown. One worker without cutaneous sporotrichosis may have had asymptomatic pulmonary sporotrichosis.

Agricultural Workers' Diseases↗

Influence of water hardness on accumulation and elimination of cadmium in two aquatic mosses under laboratory conditions.

This study investigated the effect of water hardness on the accumulation and elimination of cadmium (Cd) by two aquatic mosses, Fontinalis dalecarlica and Platyhypnidium riparioides, under laboratory conditions. The two mosses were exposed to nominal Cd concentrations of 0, 0.8, 2, and 10 microg . L-1, which includes the concentration range generally found in nature. The influence of three levels of water hardness (very soft: 11.7 mg . L-1; soft: 44.2 mg . L-1; and hard water: 92.3 mg . L-1 as CaCO3) was measured while maintaining the alkalinity and pH constant during the 28-day exposure. The Cd accumulation by the aquatic mosses was rapid, showing the potential of accumulation and the sensitivity of this biomonitor. Even if the actual Cd concentration in the water was low (concentration <0.15 microg . L-1 to 6.82 microg . L-1 of Cd), the uptake of Cd was very fast and mostly linear. This study was conducted in water hardness comparable to that found in the Canadian shield (hardness was <100 mg . L-1 as CaCO3). When the actual Cd concentration in the water was as high as 6.82 microg . L-1, the uptake of Cd was mostly linear and the steady state condition was not reach. Accumulation rates of Cd were significantly different when the mosses were in very soft (11.7 mg . L-1) as compared to hard water (92.3 mg . L-1 as CaCO3). The elimination of Cd followed a very slow process for the two species studied. The elimination rates of Cd from the mosses were not influenced by water hardness.

Bryopsida↗

A few molecules of zeaxanthin per reaction centre of photosystem II permit effective thermal dissipation of light energy in photosystem II of a poikilohydric moss.

The relationship between thermal dissipation of light energy (as indicated by the quenching of chlorophyll fluorescence), zeaxanthin availability and protonation reactions was investigated in the moss Rhytidiadelphus squarrosus (Hedw.) Warnst. In the absence of zeaxanthin and actinic illumination, acidification by 20% CO2 in air was incapable of quenching basal, so-called F0 fluorescence either in the moss or in spinach (Spinacia oleracea L.) leaves. However, 1-s light pulses given either every 40, 60 or 200 s increased thermal dissipation as indicated by F0 and Fm quenching in the presence of 20% CO2 in air in the moss, but not in spinach while reaction centres of photosystem II (PSII) were photochemically open. In the moss, a few short light pulses, which were separated by prolonged dark times, were sufficient to raise zeaxanthin levels in the presence of 20% CO2 in air. Simultaneously, quantum efficiency of charge separation in PSII was decreased. Increasing the CO2 concentration beyond 20% further decreased quantum efficiency even in the absence of short light pulses. Under conditions optimal for fluorescence quenching, one molecule of zeaxanthin per reaction centre of PSII was sufficient to decrease quantum efficiency of charge separation in PSII by 50%. Thus, in combination with a protonation reaction, one molecule of zeaxanthin was as efficient at capturing excitation energy as a photochemically open reaction centre. The data are discussed in relation to the interaction between zeaxanthin and thylakoid protonation, which enables effective thermal dissipation of light energy in the antennae of PSII in the moss but not in higher plants when actinic illumination is absent.

Bryopsida↗

The evolution of the abscisic acid-response in land plants: comparative analysis of group 1 LEA gene expression in moss and cereals.

The moss Physcomitrella patens possesses a single copy of a Group 1 LEA gene, designated PpLEA-1. Sequence analysis of the PpLEA-1 gene reveals the gene to contain a single intron in a position conserved in all members of the Group 1 LEA gene family, but also to contain a premature termination codon within the first exon. Nevertheless, a PpLEA-1 transcript accumulates in moss tissues in response both to the imposition of osmotic stress, and to the plant growth regulator abscisic acid (ABA). This response appears to be mediated at the transcriptional level, and observation of the pattern of gene expression, reported by histochemical staining of plants expressing a PpLea-1::GUS transgene suggests that the promoter responds preferentially to ABA in protonemal filaments, whereas osmotic stress induces gene expression primarily in the gametophores. Quantitative analysis of promoter activity by transient expression in Physcomitrella protoplasts shows the PpLEA-1 promoter to be highly active in response to ABA and osmotic stress. ABA-mediated transgene expression from the PpLea-1 promoter occurs at a level similar to that driven by the highly active promoter of the wheat Group 1 LEA gene, E(m). Site-directed mutagenesis of the PpLEA-1 promoter indicates that ABA-inducibility is mediated via an ACGT-core motif similar to that seen in the ABA response elements of higher plant LEA genes. However, whereas the wheat E(m )promoter is active in moss tissues, the moss promoter is not reciprocally active in cereal cells: no activity, ABA-inducible or otherwise was detected in barley aleurone protoplasts transfected with the PpLEA-1::GUS construct. We propose that ABA activation of gene expression in moss cells represents an ancestral state, with only minimal requirements for promoter recognition, whereas cereal cells require the interaction of additional factors with ABA-responsive promoters.

Abscisic Acid↗

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↗

Physcomitrella patens and Ceratodon purpureus, mosses as model organisms in photosynthesis studies.

With the discovery of targeted gene replacement, moss biology has been rapidly advancing over the last 10 years. This study demonstrates the usefulness of moss as a model organism for plant photosynthesis research. The two mosses examined in this study, Physcomitrella patens and Ceratodon purpureus, are easily cultured through vegetative propagation. Growth tests were conducted to determine carbon sources suitable for maintaining heterotrophic growth while photosynthesis was blocked. Photosynthetic parameters examined in these plants indicated that the photosynthetic activity of Ceratodon and Physcomitrella is more similar to vascular plants than cyanobacteria or green algae. Ceratodon plants grown heterotrophically appeared etiolated in that the plants were taller and plastids did not differentiate thylakoid membranes. After returning to the light, the plants developed green, photosynthetically active chloroplasts. Furthermore, UV-induced mutagenesis was used to show that photosynthesis-deficient mutant Ceratodon plants could be obtained. After screening approximately 1000 plants, we obtained a number of mutants, which could be arranged into the following categories: high fluorescence, low fluorescence, fast and slow fluorescence quenching, and fast and slow greening. Our results indicate that in vivo biophysical analysis of photosynthetic activity in the mosses can be carried out which makes both mosses useful for photosynthesis studies, and Ceratodon best sustains perturbations in photosynthetic activity.

Bryopsida↗

The use of mosses as indicators of airborne radionuclides near a major nuclear installation.

Samples of naturally growing moss (mainly Pseudoscleropodium purum and Hypnum cupressiforme ) were used to investigate the distribution of photon-emitting radionuclides, notably 129I , 137Cs, 144Ce and 241Am, at distances up to 12 km from the nuclear fuel reprocessing plant at Sellafield in West Cumbria . The relative activities of the naturally occurring radionuclides, 7Be and 210Pb, in moss and air indicated that the removal half-time of these radionuclides on moss is very long (best estimate of half-time, corrected for radioactive decay, was 240 days) and that the mosses are very efficient collectors of airborne particulates (product of deposition velocity and interception factor was 2.4 X 10(-2) ms-1). Hence the activities of artificially-produced radionuclides on the mosses present an effective record of local airborne radionuclide pollution integrated over a number of years. The activity of 210Pb in each sample was used to standardize the activities of artificial radionuclides in the samples. It was found that the activity ratio of 129I /210Pb, 137Cs/210Pb and 144Ce/210Pb declined with distance from Sellafield indicating that the major local source of these radionuclides has been material discharged to air from the site. The activity ratio of 241Am/210Pb declined with distance from the site and also with distance from the sea confirming that the resuspension of material originally discharged to sea is an important local source of 241Am.

Air Pollutants↗