Analysis of the bacterial community developing in the course of <Emphasis Type="Italic">Sphagnum</Emphasis> moss decomposition

Slow degradation of organic matter in acidic Sphagnum peat bogs suggests a limited activity of organotrophic microorganisms. Monitoring of the Sphagnum debris decomposition in a laboratory simulation experiment showed that this process was accompanied by a shift in the water color to brownish due to accumulation of humic substances and by the development of a specific bacterial community with a density of 2.4 × 10⁷ cells ml^?1. About half of these organisms are metabolically active and detectable with rRNA-specific oligonucleotide probes. Molecular identification of the components of this microbial community showed the numerical dominance of bacteria affiliated with the phyla Alphaproteobacteria, Actinobacteria, and Planctomycetes. The population sizes of the Firmicutes and Bacteroidetes, which are believed to be the main agents of bacterially-mediated decomposition in eutrophic wetlands, were low. The numbers of planctomycetes increased at the final stage of Sphagnum decomposition. The representative isolates of the Alphaproteobacteria were able to utilize galacturonic acid, the only low-molecular-weight organic compound detected in the water samples; the representatives of the Planctomycetes were able to decompose some heteropolysaccharides, which points to the possible functional role of these groups of microorganisms in the community under study. Thus, the composition of the bacterial community responsible for Sphagnum decomposition in acidic and low-mineral oligotrophic conditions seems to be fundamentally different from that of the bacterial community which decomposes plant debris in eutrophic ecosystems at neutral pH.     

Rockin' in rhythm,growth dynamics of the peat moss Sphagnum

The genus Sphagnum is an essential component in the formation and maintenance of high latitude peatlands, bogs and mires. The species grows in dense, extended mats of agglomerated shoots that allow it to retain water necessary for its growth. These mats are partly responsible for maintaining the right conditions for other species in these wetlands to thrive. In this issue of Physiologia Plantarum, Mironov et al. (2020) monitored the growth of Sphagnum riparium for a period of 4 years and revealed three distinct growth rhythms: a seasonal temperature dependent, a circalunar and a third one, synchronized with the circalunar. This synchronised nature of Sphagnum growth could contribute to its position as a key species in the maintenance of peatlands.     

Structural colour in the moss Sphagnum subnitens Russ. & Warnst

Abstract

The blue-grey sheen of dried specimens of Sphagnum subnitens is attributed to thin film interference in the walls of the hyaline cells, this being produced by reflection from upper and lower film surfaces. The reflectance is large in air and small in water, so that the colour is visible only in dried specimens. S. rubellurn lacks the blue-grey sheen because only a small part of the total area of the hyaline cells is thin enough to generate interference colour.     

Development of microsatellites for the peat moss Sphagnum capillifolium using ISSR cloning

Sphagnum mosses are major components of peat bogs but populations of many species are under threat due to habitat fragmentation resulting from the cutting of peat for fuel. We have used an intersimple sequence repeat (ISSR)‐based cloning method to develop nine polymorphic nuclear microsatellites for the peat moss species Sphagnum capillifolium. Between three and seven alleles per locus were detected in a sample of 48 haploid gametophytes and levels of gene diversity ranged from 0.5391 to 0.7960. These represent the first microsatellite markers developed for this important genus and most also exhibited cross‐species amplification across a range of common Sphagnum species.     

Transfer of fixed-N from N2-fixing cyanobacteria associated with the moss Sphagnum riparium results in enhanced growth of the moss

Background

Despite the general assumption that nitrogen fixed by associated cyanobacteria will be readily utilised for growth by the Sphagnum, no empirical evidence is available in the literature. Therefore the effects of nitrogen transfer from cyanobacteria associated with S. riparium were investigated.

Methods

Cultivation of S. riparium with and without cyanobacteria was performed under laboratory conditions for 57 days.

Results

We show that nitrogen fixation by cyanobacteria associated with Sphagnum mosses, influences moss growth by transfer of fixed nitrogen to the moss. More than 35 % of the nitrogen fixed by cyanobacteria was transferred to the newly formed moss biomass and resulted in an increase in the growth of Sphagnum biomass compared to the controls. The variation in the increase of nitrogen content explained 76 % of the biomass increment.

Conclusion

Hence, nitrogen fixation will have immediate effect on the carbon fixation by Sphagnum. This shows that factors regulating nitrogen fixation will have a direct effect on the role of Sphagnum dominated ecosystems with respect to carbon cycling.     

Experimental warming alters the community composition,diversity, and N2 fixation activity of peat moss (Sphagnum fallax) microbiomes

Sphagnum‐dominated peatlands comprise a globally important pool of soil carbon (C) and are vulnerable to climate change. While peat mosses of the genus Sphagnum are known to harbor diverse microbial communities that mediate C and nitrogen (N) cycling in peatlands, the effects of climate change on Sphagnum microbiome composition and functioning are largely unknown. We investigated the impacts of experimental whole‐ecosystem warming on the Sphagnum moss microbiome, focusing on N₂ fixing microorganisms (diazotrophs). To characterize the microbiome response to warming, we performed next‐generation sequencing of small subunit (SSU) rRNA and nitrogenase (nifH) gene amplicons and quantified rates of N₂ fixation activity in Sphagnum fallax individuals sampled from experimental enclosures over 2 years in a northern Minnesota, USA bog. The taxonomic diversity of overall microbial communities and diazotroph communities, as well as N₂ fixation rates, decreased with warming (p < 0.05). Following warming, diazotrophs shifted from a mixed community of Nostocales (Cyanobacteria) and Rhizobiales (Alphaproteobacteria) to predominance of Nostocales. Microbiome community composition differed between years, with some diazotroph populations persisting while others declined in relative abundance in warmed plots in the second year. Our results demonstrate that warming substantially alters the community composition, diversity, and N₂ fixation activity of peat moss microbiomes, which may ultimately impact host fitness, ecosystem productivity, and C storage potential in peatlands.     

North American origin and recent European establishments of the amphi-Atlantic peat moss Sphagnum angermanicum

Genetic and morphological similarity between populations separated by large distances may be caused by frequent long-distance dispersal or retained ancestral polymorphism. The frequent lack of differentiation between disjunct conspecific moss populations on different continents has traditionally been explained by the latter model, and has been cited as evidence that many or most moss species are extremely ancient and slowly diverging. We have studied intercontinental differentiation in the amphi-Atlantic peat moss Sphagnum angermanicum using 23 microsatellite markers. Two major genetic clusters are found, both of which occur throughout the distributional range. Patterns of genetic structuring and overall migration patterns suggest that the species probably originated in North America, and seems to have been established twice in Northern Europe during the past 40,000 years. We conclude that similarity between S. angermanicum populations on different continents is not the result of ancient vicariance and subsequent stasis. Rather, the observed pattern can be explained by multiple long-distance dispersal over limited evolutionary time. The genetic similarity can also partly be explained by incomplete lineage sorting, but this appears to be caused by the short time since separation. Our study adds to a growing body of evidence suggesting that Sphagnum, constituting a significant part of northern hemisphere biodiversity, may be more evolutionary dynamic than previously assumed.     

Sphagnum production and decomposition in a restored cutover peatland

Natural peatlands represent a long-termsink of atmospheric carbon dioxide(CO₂), however, drained and extractedpeatlands can represent a source ofatmospheric CO₂. The restoration ofSphagnum mosses on abandoned milledpeatlands has the potential to sequesteratmospheric CO₂ thereby returning thepeatland to a peat accumulating system.Micrometeorological and chambermeasurements of net ecosystem CO₂exchange are proven methods forinvestigating production and decompositionprocesses in both natural, extracted, andrestored peatlands. However, this approachis relatively expensive because ofinfrastructure and human resources that notonly limits potential use for ecologicalmanagers but it limits the number of sitesthat can be monitored due to high spatialvariability. Here we present crank wire anddestructive sampling productionmeasurements, litter bag decompositionmeasurements and measurements of netecosystem CO₂ exchange made in arestored peatland and natural peatlandsites nearby. The objectives were to assessproduction and decomposition rates in thetwo systems as well as to compare thedifferent measurements techniques.Estimates of Sphagnum fuscumproduction at a restored peatland, usingthe different methods, followed the trend:crank wire < destructive sampling < gasexchange, with the two last methodsproviding comparable estimates. Productionestimates using crank wires in cutover peatsurfaces with a thin newly formed Sphagnum mat were shown unreliable due topeat subsidence. Results using thedestructive sampling method suggest thatSphagnum production varies betweenspecies (S. fuscum > S.capillifolium) according to their abilityto withstand harsh conditions on restoredpeat surfaces. Decomposition rate was alsosignificantly greater (p<0.05) for S. capillifolium than S. fuscum,resulting in an overall plant accumulationgreater for S. fuscum. Although therestored surfaces were fairly young,production rates estimated on cutoversurfaces that were fully covered with athin Sphagnum mat compared withproduction rates observed in natural sitesnearby.     

Freezing cytorrhysis and critical temperature thresholds for photosystem II in the peat moss Sphagnum capillifolium

Leaflets of Sphagnum capillifolium were exposed to temperatures from ?5°C to +60°C under controlled conditions while mounted on a microscope stage. The resultant cytological response to these temperature treatments was successfully monitored using a light and fluorescence microscope. In addition to the observable cytological changes during freezing cytorrhysis and heat exposure on the leaflets, the concomitant critical temperature thresholds for inactivation of photosystem II (PS II) were studied using a micro fibre optic and a chlorophyll fluorometer mounted to the microscope stage. Chlorophyllous cells of S. capillifolium showed extended freezing cytorrhysis immediately after ice nucleation at ?1.1°C in the water in which the leaflets were submersed during the measurement. The occurrence of freezing cytorrhysis, which was visually manifested by cell shrinkage, was highly dynamic and was completed within 2 s. A total reduction of the mean projected diameter of the chloroplast containing area during freezing cytorrhysis from 8.9 to 3.8 μm indicates a cell volume reduction of approximately ?82%. Simultaneous measurement of chlorophyll fluorescence of PS II was possible even through the frozen water in which the leaf samples were submersed. Freezing cytorrhysis was accompanied by a sudden rise of basic chlorophyll fluorescence. The critical freezing temperature threshold of PS II was identical to the ice nucleation temperature (?1.1°C). This is significantly above the temperature threshold at which frost damage to S. capillifolium leaflets occurs (?16.1°C; LT₅₀) which is higher than observed in most higher plants from the European Alps during summer. High temperature thresholds of PS II were 44.5°C which is significantly below the heat tolerance of chlorophyllous cells (49.9°C; LT₅₀). It is demonstrated that light and fluorescence microscopic techniques combined with simultaneous chlorophyll fluorescence measurements may act as a useful tool to study heat, low temperature, and ice-encasement effects on the cellular structure and primary photosynthetic processes of intact leaf tissues.     

Major and trace elements in Sphagnum moss from four southern German bogs,and comparison with available moss monitoring data

In this paper, we present concentrations of an array of major and trace elements (Ag, Al, As, Ba, Bi, Cd, Co, Cr, Cu, Fe, Mn, Mo, Rb, Sb, Sc, Sr, Th, Tl, U, V, Zn) in living Sphagnum mosses from four southern German bogs and compare them with moss monitoring data of the respective regions. To do this, Sphagnum mosses were collected in Upper Bavaria (Oberbayern, OB) and the Northern Black Forest (Nordschwarzwald, NBF). Surfaces of Sphagnum carpets were marked with plastic mesh and, one year later, the annual moss production was harvested. Up to 12 samples (40 cm × 40 cm) were collected per site, and 6–10 sites investigated per bog. The concentrations of these elements were then determined in acid digests using sector field ICP-MS. Variations within a given sampling site were in the range of 2 to 3-fold for all major and trace element concentrations except for Mn (12-fold) and Tl (38-fold). For most of the elements, concentrations between bogs of a given region were significantly different and atmospheric deposition of particles seems to be considerably affected by local circumstances such as tree canopy interception and microtopography. Comparing trace element concentrations measured in Sphagnum mosses for 2007 with published moss monitoring data for 2005 resulted in a very good agreement for most elements. Clearly, Sphagnum mosses from bogs are useful biomonitors for estimating atmospheric contamination by metals. This supports the use of Sphagnum in atmospheric deposition monitoring especially in cases where Sphagnum is abundant (e.g., boreal forests). In regions with neither bogs nor forests, living Sphagnum moss bags could be used to the same effect.     

The narrow endemic Norwegian peat moss Sphagnum troendelagicum originated before the last glacial maximum

It is commonly found that individual hybrid, polyploid species originate recurrently and that many polyploid species originated relatively recently. It has been previously hypothesized that the extremely rare allopolyploid peat moss Sphagnum troendelagicum has originated multiple times, possibly after the last glacial maximum in Scandinavia. This conclusion was based on low linkage disequilibrium in anonymous genetic markers within natural populations, in which sexual reproduction has never been observed. Here we employ microsatellite markers and chloroplast DNA (cpDNA)-encoded trnG sequence data to test hypotheses concerning the origin and evolution of this species. We find that S. tenellum is the maternal progenitor and S. balticum is the paternal progenitor of S. troendelagicum. Using various Bayesian approaches, we estimate that S. troendelagicum originated before the Holocene but not before c. 80 000 years ago (median expected time since speciation 40 000 years before present). The observed lack of complete linkage disequilibrium in the genome of this species suggests cryptic sexual reproduction and recombination. Several lines of evidence suggest multiple origins for S. troendelagicum, but a single origin is supported by approximate Bayesian computation analyses. We hypothesize that S. troendelagicum originated in a peat-dominated refugium before last glacial maximum, and subsequently immigrated to central Norway by means of spore flow during the last thousands of years.     

Biomagnetic monitoring of urban air pollution using moss bags (Sphagnum girgensohnii)

Active moss biomonitoring is widely applied in polluted areas for monitoring of airborne particle-bound trace element pollution. This study explored the suitability of the moss Sphagnum girgensohnii for biomagnetic monitoring in cities. To this end S. girgensohnii moss bags were exposed at three different microenvironments characterized by heavy traffic – street canyons, a city tunnel and parking garages during the summer and autumn of 2011 in the city of Belgrade. The ferro(i)magnetic PM fraction in the moss samples was quantified by Saturated Isothermal Remanent Magnetization (SIRM) and the measured values were compared with the trace element concentration in the moss samples. SIRM values were significantly different across the considered urban microenvironments. Moreover, a high correlation between moss SIRM values and concentrations of Al, Ba, Co, Cr, Cu, Fe, Ni and Pb was found. These results demonstrate that moss bags can be effectively applied for biomagnetic monitoring of the spatio-temporal distribution of road traffic and vehicles derived pollutants in urban areas.     

Establishment and growth characterization of suspension culture of cells from the moss, Sphagnum imbrication

A green-pigmented callus of the moss, Sphagnum imbricatum Hornsch. ex. Russ., was induced and a chlorophyllous cell suspension culture was established using a modified Murashige and Skoog's medium without plant hormones. Cell growth in the light in the presence of glucose started after a short lag and was exponential for 12 days. The chlorophyll level was about 15 μg (mg cell dry weight)⁻¹ and photosynthetic activity ca 20 to 50 μmol O₂ (mg chlorophyll)⁻¹h⁻¹. Cell growth in the light was negligible in the absence of glucose under ordinary air, but photoautotrophic growth was possible under elevated CO₂ concentrations. In the dark, the moss cells grew heterotrophically and continued to synthesize chlorophyll, although at a much reduced rate. The suspension-cultured cells redifferentiated protonemata and shoots when transferred to solid Knop's medium. In contrast to the callus cells, which could not assimilate nitrate, redifferentiated plantlets could use nitrate as the sole nitrogen source.     

Photosynthesis, chlorophyll fluorescence and spectral reflectance in Sphagnum moss at varying water contents

Moss samples from the Fluxnet-Canada western peatland flux station in the Boreal Region of Alberta were measured in the laboratory to obtain the net photosynthesis rate and chlorophyll fluorescence of the moss under controlled environmental conditions, including the regulation of moss water content, simultaneously with measurements of moss spectral reflectance. One objective was to test whether the photochemical reflectance index (PRI) detected changes in moss photosynthetic light-use efficiency that were consistent with short-term (minutes to hours) changes in xanthophyll cycle pigments and associated changes in non-photochemical quenching (NPQ), as recorded by chlorophyll fluorescence. The rate of net photosynthesis was strongly inhibited by water content at values exceeding approximately 9 (fresh weight/dry weight) and declined as the water content fell below values of approximately 8. Chlorophyll fluorescence measurements of maximum photosystem II efficiency generally remained high until the water content was reduced from the maximum of about 20 to values of approximately 10–11, and then declined with further reductions in moss water content. A significant linear decline in NPQ was observed as moss water content was reduced from maximum to low water content values. There was a strong negative correlation between changes in NPQ and PRI. These data suggest that PRI measurements are a good proxy for short-term shifts in photosynthetic activity in Sphagnum moss. A second objective was to test how accurately the water band index (WBI, ratio of reflectance at 900 and 970 nm) recorded changes in moss water content during controlled laboratory studies. Strong linear relationships occurred between changes in moss water content and the WBI, although the slopes of the linear relationships were significantly different among sample replicates. Therefore, WBI appeared to be a useful tool to determine sample-specific water content without destructive measurements.     

Analysis of the bacterial community in glassy-winged sharpshooter heads

The glassy‐winged sharpshooter (GWSS), Homalodisca vitripennis, is an important vector of various strains of Xylella fastidiosa, which cause disease in a variety of economically important plants. These diseases include citrus variegated chlorosis, oleander leaf scorch and Pierce's Disease of grapevines. Symbiotic control (SC) is a new strategy that uses symbiotic endophytes as biological control agents to antagonize or displace the pathogenic strains of X. fastidiosa. Candidate endophytes for use in SC must occupy the xylem of host plants and attach to the pre‐cibarium and cibarium of sharpshooter insects in order to have access to the pathogen. The study of the bacterial community of GWSS heads by isolation and denaturing gradient gel electrophoresis (DGGE) revealed the presence of species that may be suitable for use in SC. In addition, the results indicated that two important factors, insect age and choice of host plant, affect the composition of the bacterial community in GWSS heads. The main bacterial genera isolated as colonizers of GWSS heads were identified, using partial 16S rRNA gene sequencing, as Bacillus, Pseudomonas, Pedobacter and Methylobacterium, as well as the species Curtobacterium flaccumfaciens. DGGE patterns revealed a diversity of endophytic species able to colonize the GWSS head. The main genera isolated in culture were also identified using this technique. Principal component analysis (PCA) from polymerase chain reaction (PCR)‐DGGE patterns indicated that the bacteria inhabiting the GWSS head are similar to those found as endophytes inside the host plants, and that insect developmental stage and preferential feeding on one host plant species over another are important factors in determining the composition of the bacterial community in the GWSS head. However, a shift in host plants for a small period of time did not cause changes in the compositions of these communities.     

Microclimate and production of peat moss Sphagnum palustre L. in the warm‐temperate zone

Sphagnum palustre L. is one of the few Sphagnum species distributed in the warm‐temperate zone. To elucidate the mechanisms that enable S. palustre to maintain its productivity under warm climatic conditions, we examined the temperature conditions and photosynthetic characteristics of this species in a lowland wetland in western Japan. Moss temperatures during the daytime were much lower than the air temperature, particularly during summer. The optimum temperature for the net photosynthetic rate was approximately 20°C, irrespective of the season, but summer and autumn samples maintained high rates at higher temperatures as well. The net photosynthetic rate at near light saturation was much higher during summer–autumn than during spring–winter. A model estimation in which net production was calculated from the photosynthetic characteristics and microclimatic data showed that both the low temperature of the moss colony and the seasonal shift in photosynthetic characteristics are among the mechanisms that enable this species to maintain its productivity under warm climatic conditions.     

Testate amoebae community structure in the moss biotopes of streams

The specific composition and community structure of testate amoebae in the moss biotopes of streams were studied in the Sura River basin (Middle Volga region). Twenty-nine species and forms were identified. The eurybiont species Trinema enchelys, Euglypha ciliata glabra, Centropyxis aerophila, Tracheleuglypha dentata, Crythion dubium, Centropyxis cassis, Trinema complanatum, and Trinema lineare are dominant. The species richness varies from 2 to 11 species per sample, the abundance is 100 to 4000 ind./g of absolutely dry moss. The degree of forest covering in the basin, hydrochemical peculiarities (the amount of biogenic elements and water hardness) of streams, the size of streams, and environmental contamination are possible factors that determine the specific character of the composition and structure of testate amoebae communities.     

Peat respiration and decomposition in Antarctic terrestrial moss communities

Oxygen uptake by the peat of two Antarctic bryophyte communities (a moss turf and a moss carpet) is converted to organic matter loss and used to derive the rate of decomposition. The decay rates obtained in this way are evaluated in two mathematical models which simulate the accumulation of dead organic matter (DOM) in the communities from the litter production and decomposition rate. Litter production, the extent of DOM accumulations at present on the sites and mean decomposition rates (i.e. fraction of standing crop lost per year) were 409 g m^-2 year^-1, 33.5 kg m^-2 and 0.017 g g^-1 year^-1 in the moss turf and 392 g m^-2 year^-1, 29.6 kg m-² and 0.010 g g^-1 year^-1 in the moss carpet respectively (all weights expressed as dry weight). Aerobic decomposition rate declined with depth in both communities. From the model's predictions it is suggested that the observed decay rate was too high in the moss turf and too low in the carpet. Possible reasons for this are discussed and suggestions made for future work.     

南极三洋藓(Sanionia uncinata)细菌群落组成分析

【背景】南极苔藓中蕴藏着丰富的极地微生物资源,然而目前对南极苔藓细菌群落组成的研究较少,限制了对这类极端生境微生物资源的开发和应用。【目的】在南极长城站不同地点随机取样,揭示南极三洋藓(Sanioniauncinata)的细菌群落结构组成。【方法】采用IlluminaHiSeq高通量测序技术对3个苔藓样品的16S rRNA基因V4区进行测序及生物信息学分析。【结果】3个苔藓样品中共获得细菌总优化序列273 367条。基于97%序列相似度,细菌优化序列聚类为9 579个OTU。分类地位明确的细菌可归为14门27纲50属。优势类群是变形菌门(Proteobacteria)(30.70%)、拟杆菌门(Bacteroidetes)(19.67%)、疣微菌门(Verrucomicrobia)(12.43%)、蓝细菌门(Cyanobacteria)(10.55%)及放线菌门(Actinobacteria)(9.36%)。在属水平上,56.73%属于未知细菌。【结论】南极三洋藓(Sanionia uncinata)中具有丰富多样的细菌,该研究为今后极地微生物研究奠定了基础。