Sphagnum Species Research Articles

Drought mediates Sphagnum defense response to herbivory.

The expected concomitant increase in multiple stressors such as herbivory and drought may threaten peatland ecosystems. How Sphagnum, the ecological engineers of peatlands, responds to combined stressors remains largely unexplored. Here we aimed to clarify resource allocations in Sphagnum during concomitant herbivory and drought. S. magellanicum and S. fuscum were exposed to drought and herbivory together or separately in laboratory experiments and analyzed for growth (biomass production and net photosynthetic rate), defense (phenolics in leachates and phenolics in extraction) and nonstructural carbohydrates (soluble sugar and starch) in relation to untreated controls. Herbivory and drought had significant interactive effects on Sphagnum growth and defense. In both species, drought without herbivory reduced the phenolics in leachate, but with herbivory increased phenolics, indicating a synergistic effect between herbivory and drought on Sphagnum defense. Both stressors significantly decreased biomass production, with the combined stress having a more negative effect. Interestingly, a growth-defense trade-off was found in the drought treatment of both Sphagnum species, but disappeared in the wet treatment. Conversely, a trade-off between soluble sugars and phenolics was found in the wet but not in the drought treatment, suggesting that soluble sugars may play a role in inducing the defense and hence mask the growth-defense trade-off in peat mosses. Our results emphasize that predicting the impact of combined stressors on peat moss traits is complex and challenging. Future models should account for the effects of multiple environmental stressors to guide peatland conservation under climate warming.

Comparative Photosynthetic Capacity, Respiration Rates, and Nutrient Content of Micropropagated and Wild-Sourced Sphagnum

The rapid, effective restoration of degraded peatlands is urgently needed to reduce their current high levels of carbon loss. The re-introduction of Sphagnum moss, along with re-wetting, is key to returning carbon sequestration and retention capabilities to northern degraded bogs. Micropropagated Sphagnum has already been applied in large quantities, and more is planned, for restoration projects in Britain and parts of Europe. A comparison with wild-sourced Sphagnum material is therefore pertinent to demonstrate its safety and suitability for wide-scale application. Six Sphagnum species of both micropropagated and wild-sourced origin were assessed for photosynthetic capacity, nutrient content, form parity, chlorocyst size, and chloroplast numbers. Micropropagated Sphagnum had significantly higher light-saturated photosynthesis (Pmax) rates, little color expression, an open growth habit, greater chloroplast numbers, and more numerous, smaller shoot apices than wild-sourced Sphagnum. Higher Pmax rates were associated with a lower bulk density and higher tissue nutrient concentrations. Potentially, greater chloroplast numbers in micropropagated Sphagnum facilitate higher photosynthesis rates, driving rapid growth in early-stage plants, particularly in optimum moisture conditions. Micropropagated Sphagnum can be used confidently, propagated in large quantities, and will likely establish well on application to sites where re-wetting has already occurred, therefore making it highly beneficial for the restoration of degraded bogs.

Pigment Complex, Growth and Chemical Composition Traits of Boreal Sphagnum Mosses (Mire System "Ilasskoe", North-West of European Russia).

Sphagnum mosses play a significant role in peat formation and carbon sequestration in mire ecosystems. It is critical to investigate the productivity and chemical composition of different Sphagnum species in order to assess their role in the global carbon cycle and potential in light of climate change. The data on productivity and growth characteristics during the growing season, group chemical composition and elemental composition at the beginning and end of the growing season, as well as aspects of the pigment complex operation, were collected for four Sphagnum species: Sphagnum lindbergii Schimp., S. fuscum (Schimp.) Klinggr., S. divinum Flatberg & K. Hassel, and S. squarrosum Crome. High cover density and productivity, low ability to decompose, and constancy of the pigment complex of S. fuscum reflect a high degree of adaptation to the specific conditions of ridges. A constant chemical composition of S. lindbergii during the growing season can be explained by stable conditions of hollows that allow it to maintain its metabolic processes, but the light conditions in hollows bring the reaction of the pigment apparatus of this species closer to shaded S. divinum and S. squarrosum. S. lindbergii and S. squarrosum contain more nitrogen than other species and have a greater ability to decompose.

SEASONAL CHANGES IN THE CONCENTRATION AND RATIOS OF PHOTOSYNTHETIC PIGMENTS IN THE THREE SPHAGNUM SPECIES

Background. There is reason suppose that the concentration of photosynthetic pigments in sphagnum mosses, as in vascular plants, changes throughout the year and varies from species to species. Moreover, hollow sphagnum species have a higher growth and photosynthesis rate than ridge sphagnum species, which suggests the presence of differences in the pigment complex of species from different habitats. Purpose. Evaluation of quantitative changes in the pigment complex of the sphagnum mosses growing in different microclimatic conditions within oligotrophic bogs during the growing season at high latitudes. Materials and methods. The content of photosynthetic pigments in the three Sphagnum species growing in oligotrophic conditions was determined using the photometric method in an acetone extract once a month from May to October. The study was carried out for S. fuscum and S. lindbergii in 2021-2022, and for S. angustifolium only in 2022. Statistical data processing was carried out in Microsoft Excel 2010 and SPSS Statistics 11. Results. Seasonal and interannual variability in the content of chlorophylls, carotenoids, and their ratios was revealed for the studied sphagnum species. The pigment complex of S. lindbergii and S. angustifolium during the growing season is influenced by the bog water level and air temperature. Changes in the pigment complex of S. fuscum are associated with the amount of precipitation. Conclusion. Apparently, the seasonal and interannual dynamics of the pigment complex of sphagnum mosses is species-specific, but to further verify this statement it is necessary to study other species of sphagnum mosses from the same habitats, since the dynamics may be associated not so much with a peculiar species, but with the ecological conditions of the habitat.

The water absorption capacity of 21 Sphagnum species

Sphagnum moss (peat moss) is the dominant land plant in peatlands, and it plays an essential role in peatland ecosystems because its remarkable water-holding capacity helps to conserve water resources and maintain the anoxic environment of peatlands. Sphagnum moss exhibits a super ability to absorb water. However, the water absorption capacity of different species and different parts of the same species is not well understood. In this study, we measured the short-term and saturated water absorption of 21 Sphagnum species from China. The results showed that the water absorption capacity of the investigated peat mosses was much higher than that of other bryophytes. Sphagnum imbricatum had the highest saturated water absorption capacity (about 44 times its dry weight), while the peat moss with the lowest saturated water absorption was Sphagnum flexuosum (about 19 times its dry weight). In addition, we determined the relative volume of hyalocysts of the spreading and pendent branch leaves of 18 Sphagnum species and further measured the water absorption capacity of the pendent branch, spreading branch, and stem of six of them. The results revealed that both pendent and spreading branches had strong water absorption capacity and were the primary water-absorbing parts of peat mosses. The water-absorbing capacity of different Sphagnum species was linearly related to the volume of hyalocysts. The results provide an essential scientific basis for selecting high-quality germplasm resources of peat moss.

Variations in water economy traits in two Sphagnum species across their distribution boundaries.

We assessed changes in traits associated with water economy across climatic gradients in the ecologically similar peat mosses Sphagnum cuspidatum and Sphagnum lindbergii. These species have parapatric distributions in Europe and have similar niches in bogs. Sphagnum species of bogs are closely related, with a large degree of microhabitat niche overlap between many species that can be functionally very similar. Despite this, ecologically similar species do have different distributional ranges along climatic gradients that partly overlap. These gradients may favor particular Sphagnum traits, especially in relation to water economy, which can be hypothesized to drive species divergence by character displacement. We investigated traits relevant for water economy of two parapatric bryophytes (Sphagnum cuspidatum and S. lindbergii) across the border of their distributional limits. We included both shoot traits and canopy traits, i.e., collective traits of the moss surface, quantified by photogrammetry. The two species are ecologically similar and occur at similar positions along the hydrological gradient in bogs. The biggest differences between the species were expressed in the variations of their canopy surfaces, particularly surface roughness and in the responses of important traits such as capitulum mass to climate. We did not find support for character displacement, because traits were not more dissimilar in sympatric than in allopatric populations. Our results suggest that parapatry within Sphagnum can be understood from just a few climatic variables and that climatic factors are stronger drivers than competition behind trait variation within these species of Sphagnum.

The Sphagnum Species of Massachusetts

The Sphagnum Species of Massachusetts

Variation in Water-Holding Capacity in Sphagnum Species Depends on Both Plant and Colony Structure.

Peatlands have become a focal point in climate mitigation strategies as these ecosystems have significant carbon sequestration capacities when healthy but release CO2 and other greenhouse gases when damaged. However, as drought episodes become more frequent and prolonged, organisms key to the functioning of some peatlands are increasingly under pressure from desiccation. The Sphagnum mosses, which tend to keep their ecosystem waterlogged and many of whom promote peat formation, are only mildly desiccation-tolerant in comparison to other mosses. The role of Sphagnum anatomy and colony structure is poorly understood in the context of desiccation resilience. Using four different Sphagnum species belonging to four different subgenera and positions along the gradient of the water table, we show that plant morphological traits and colony density are important determinants of water storage capacity. Our results show that, as previously postulated, the majority of the water is stored in an easily exchangeable form, probably extracellularly, and that plant morphological traits, specifically the type and presence of branches, are major contributors to water storage and can explain some of the interspecies variation. We also show that plant density is another important determinant for water storage capacity as higher densities hold larger quantities of water per unit of biomass for all four species, which increases resilience to desiccation. The results presented here suggest that species choice and planting density should receive more attention when considering peatland restoration strategies.

Checklists for the Sphagnum (Sphagnaceae) species of New England, New York and New Jersey

Checklists for the Sphagnum (Sphagnaceae) species of New England, New York and New Jersey

A deepened water table increases the vulnerability of peat mosses to periodic drought

Abstract Here we address the combined impact of multiple stressors that are becoming more common with climate change. To study the combined effects of a lower water table (WT) and increased frequency of drought periods on the resistance and resilience of peatlands, we conducted a mesocosm experiment. This study evaluated how the photosynthesis of lawn Sphagnum mosses responds to and recovers from an experimental periodic drought after exposure to the stresses of a deep or deepened WT (naturally dry and 17‐year‐long water level drawdown [WLD] in fen and bog environments). We aimed to quantify if deep WTs (1) support acclimation to drought, or (2) increase the base‐level physiological stress of mosses or (3) exacerbate the impact of periodic drought. There was no evidence of acclimation in mosses from drier environments; periodic drought decreased the photosynthesis of all Sphagnum species studied. WLDdecreased the photosynthesis of bog‐originating mosses prior to periodic drought, indicating that these mosses were stressed by the hydrological change. Deep WTs exacerbated Sphagnum vulnerability to periodic drought, indicating that the combination of drying habitats and increasing frequency of periodic drought could lead to a rapid transition in lawn vegetation. Water‐retaining traits may increase Sphagnum resilience to periodic drought. Large capitula size was associated with a higher resistance; the bog originating species studied here lacked large capitula or dense carpet structure and were more vulnerable to drought than the larger fen originating species. Consequently, lawns in bogs may become threatened. Recovery after rewetting was significant for all mosses, but none completely recovered within 3 weeks. The most drought‐resilient species had fen origin, indicating that fens are less likely to undergo a sudden transition due to periodic drought. Synthesis: Water level drawdown associated with climate change increases the sensitivity of Sphagnum mosses to periods of drought and moves them closer to their tipping point as species on the edge of their ecological envelope rapidly shut down photosynthesis and recover poorly.

Moss removal facilitates decomposition and net nitrogen loss of monospecific and mixed-species litter in a boreal peatland

AbstractLitter decomposition plays an important role in biogeochemical cycling in boreal peatlands, where mosses, especially Sphagnum species, are a determinant. In recent decades, these peatlands have experienced a decline in moss cover due to abrupt climate warming and atmospheric nitrogen (N) deposition. To reveal the effect of the reduction in moss cover on litter decomposition, we adopted a field living moss removal experiment (with the senesced tissues remaining) in a Sphagnum-dominated boreal peatland, and investigated litter mass loss and net N loss of three deciduous woody species decomposing in monocultures and mixtures over 3 years. Based on the observed and predicted mass loss and net N loss of litter mixtures, we divided litter mixing effects into additive (no significant difference), synergistic (observed value greater than predicted value), and antagonistic (observed value lower than predicted value) effects. Across 3 years of decomposition, moss removal increased litter mass loss and net N loss, irrespective of single- or mixed-species compositions. Moss removal generally changed litter mixing effects on mass loss from antagonistic to additive effects in the initial 2 years, but from synergistic to additive effects after 3 years of decomposition. Regarding net N loss of litter mixtures, moss removal often resulted in a shift from additive to synergistic effects or from antagonistic to additive effects after 2 and 3 years of decomposition. Our observations suggest that the declines in living moss cover can accelerate litter decomposition and nutrient release, and highlight that living moss loss makes litter mixture decomposition predictable by reducing non-additive effects in boreal peatlands. Given the widespread occurrence of reduced moss cover in boreal peatlands, the mechanisms explaining living moss controls on litter decomposition and N cycling should receive significant attention in further studies.

A hidden herbivory effect on Sphagnum reproduction.

Defence theories provide predictions about trade-offs in the allocation of resources to defence and growth. However, very little is known about how pressure from herbivores influences the allocation of resources during reproduction. Two common peatland bryophyte species, Sphagnum angustifolium and S. capillifolium, were chosen as study species. Vegetative and reproductive shoots of both Sphagnum species were subjected to treatments with and without herbivores in a lab experiment. After 4 weeks of exposure to herbivores in a growth chamber, we measured biomass production, net photosynthesis rate, defence traits (phenolics in leachate and phenolics in extract), nonstructural carbohydrates (soluble sugar and starch), and reproductive traits (capsule number, weight and diameter, and spore germination) of both Sphagnum species. Reproductive shoots had higher constitutive defence than vegetative shoots in S. angustifolium, and a similar pattern was observed in S. capillifolium. With herbivory, reproductive shoots showed stronger induced defence (released more phenolics) than vegetative shoots in S. capillifolium, but not in S. angustifolium. Herbivory had no effect on capsule number, weight, or diameter, but reduced spore germination percentage by more than half in both species. Our study highlights the hidden effects of herbivory on reproduction of Sphagnum and indicates the presence of maternal effects in bryophytes. Ecologists will benefit from examining both quality- and quantity-based traits when attempting to estimate the herbivory effect on plant fitness.

The impact of drying on the structure and photosynthesis of boreal peatland vegetation

Boreal peatlands harbour large stores of carbon as peat below their surfaces. Climate change is expected to cause drying in northern peatlands, which will in turn impact the carbon balance of these ecosystems that is maintained by high water tables and the hydrologically sensitive plants growing there. This study aims to quantify how vegetation will be structured (I) and photosynthesize (II, III) in a future climate as emulated by long-term water level drawdown (WLD). To do this, changes in the vegetation and its photosynthesis after WLD are linked, and the response of Sphagnum mosses to periodic drought is investigated. Field measurements were done at a long-term WLD field experiment that contained a rich (mesotrophic) fen, a poor (oligotrophic) fen and a bog (ombrotrophic) site. Measurements included vegetation surveys from existing permanent sample plots and leaf-level carbon dioxide exchange measurements. For an experiment in controlled conditions peatland surface cores from this field experiment were transported to a greenhouse where the photosynthesis of lawn Sphagna during and after an experimental periodic drought was measured. The field study revealed that the response of peatland vegetation to WLD depend on peatland type. The species composition in the rich fen was the most impacted by WLD, while the bog vegetation demonstrated stability. Similarly, large increases in photosynthesis occurred following WLD on the vascular plant-covered rich fen, while changes were negligible on the Sphagnum-carpeted bog. The vegetation on the two fens shifted from an open sedge-, or sedge and Sphagnum-dominated ecosystem, to a tree-dominated ecosystem. Canopy development following WLD further accelerated vegetation changes by shading and sheltering the understorey vegetation. Vascular plants were the most likely to increase productivity from WLD as they are best suited to utilize the nutrients made available by peat mineralization, while Sphagnum moss photosynthesis was impacted little. The greenhouse study revealed that lawn Sphagnum mosses exposed to long-term WLD were more vulnerable to drought compared to those from wet sites. Large capitula typical to fen Sphagnum species appeared to be beneficial for surviving periodic drought. This work demonstrated that climate change as emulated by long-term WLD will have a large impact on the vegetation composition of northern peatlands and increase photosynthetic function of these ecosystems, fens in particular. To better predict climate feedbacks from these changes, carbon dynamics including peatland vegetation dynamics should be updated in global process models. Future research to better understand the tipping point of different peatland types after WLD in different climatic regions will help us to predict changes in these diverse and globally important systems.

Contrasting potential for biological N2 fixation at three polluted central European Sphagnum peat bogs: combining the 15N2-tracer and natural-abundance isotope approaches

Abstract. Availability of reactive nitrogen (Nr) is a key control on carbon (C) sequestration in wetlands. To complement the metabolic demands of Sphagnum in pristine rain-fed bogs, diazotrophs supply additional Nr via biological nitrogen fixation (BNF). As breaking the triple bond of atmospheric N2 is energy-intensive, it is reasonable to assume that increasing inputs of pollutant Nr will lead to BNF downregulation. However, recent studies have also documented measurable BNF rates in Sphagnum-dominated bogs in polluted regions, indicating the adaptation of N2 fixers to changing N deposition. Our aim was to quantify BNF in high-elevation peatlands located in industrialized central Europe. A 15N2-tracer experiment was combined with a natural-abundance N-isotope study at three Sphagnum-dominated peat bogs in the northern Czech Republic in an attempt to assess the roles of individual BNF drivers. High short-term BNF rates (8.2 ± 4.6 g N m2 d−1) were observed at Malé mechové jezírko, which receives ∼ 17 kg Nr ha−1 yr−1. The remaining two peat bogs, whose recent atmospheric Nr inputs differed from Malé mechové jezírko by only 1–2 kg ha−1 yr−1 (Uhlír̆ská and Brumiště), showed zero BNF. The following parameters were investigated to elucidate the BNF difference: the NH4+-N / NO3--N ratio, temperature, wetness, Sphagnum species, organic-N availability, possible P limitation, possible molybdenum (Mo) limitation, SO42- deposition, and pH. At Malé mechové jezírko and Uhlír̆ská, the same moss species (S. girgensohnii) was used for the 15N2 experiment; therefore, the host identity could not explain the difference in BNF at these sites. Temperature and moisture were also identical in all incubations and could not explain the between-site differences in BNF. The N : P stoichiometry in peat and bog water indicated that Brumiště may have lacked BNF due to P limitation, whereas non-detectable BNF at Uhlír̆ská may have been related to the 70-fold higher SO42- concentration in bog water. Across the sites, the mean natural-abundance δ15N values increased in the following order: atmospheric deposition (−5.3 ± 0.3 ‰) < Sphagnum (−4.3 ± 0.1 ‰) < bog water (−3.9 ± 0.4 ‰) < atmospheric N2 (0.0 ‰). Only at Brumiště was N in Sphagnum significantly isotopically heavier than in atmospheric deposition, possibly indicating a longer-term BNF effect. Collectively, our data highlight spatial heterogeneity in BNF rates under high Nr inputs as well as the importance of environmental parameters other than atmospheric Nr pollution in regulating BNF.

Sphagnum divinum Flatberg & K. Hassel ve Sphagnum girgensohnii Russow (Sphagnopsida)'nin Fitokimyasal İçeriklerinin ve Antioksidan Aktivitelerinin Belirlenmesi

This study aimed to explore the phytochemical constituents and antioxidant activity of methanol extracts from Sphagnum divinum and Sphagnum girgensohnii. Screening methods were employed to identify the phytochemical groups present in the methanol extracts of these two Sphagnum species, which have been relatively understudied for their antioxidant potential. The antioxidant properties of the extracts were evaluated using in vitro DPPH, CUPRAC, and phosphomolybdate assays. The experimental results revealed that the methanol extract of S. divinum exclusively contained phenols and tannins, while the methanol extract of S. girgensohnii contained phenols, tannins, and saponins. Alkaloids and flavonoids were not detected in either bryophyte. The methanol extracts of both S. divinum and S. girgensohnii exhibited DPPH scavenging activity exceeding 50% at all tested concentrations. An increase in DPPH scavenging ability was observed with increasing extract concentration. The DPPH scavenging activity of S. divinum methanol extract was consistently higher than that of S. girgensohnii methanol extract across all tested concentrations. These findings suggest that S. divinum and S. girgensohnii hold promise as potential sources of antioxidant compounds.

Monthly dynamics of phenolic release and allelopathic effect in hollow and hummock Sphagnum

Monthly variation in accumulation and release of phenolics in bryophytes, and their allelopathic effects are little known despite their ecological significance. We conducted a field transplant experiment to investigate Sphagnum growth and extracellular enzymes that modulate phenolic contents and release along water table level (WTL) gradient. Specifically, we assessed monthly changes in height increment, phenolic release, and allelopathic effect during the growing season, as well as extracellular enzymes at the end of the experiment, for the two Sphagnum species: Sphagnum angustifolium C.E.O. Jensen, 1896, a hollow dwelling species, and Sphagnum magellanicum Bridel, 1798, a hummock species. Both Sphagnum species showed season- and WTL-dependent height increment and released phenolics. Sphagnum angustifolium exhibited negative allelopathy regardless of WTL, while S. magellanicum demonstrated mainly positive allelopathy at high WTL and negative allelopathy at low WTL. The degree of allelopathy was not contingent on phenolic release, but phenolic production was negatively correlated with height growth of the two species at their atypical habitats. Moreover, phenoloxidase or peroxidase did not show high activity at low WTL compared to high WTL. Our findings demonstrate for the first time that the strength and direction of allelopathy in Sphagnum may vary depending on the growing precipitation and temperature, WTL, and inter- and intraspecific trait variability. Longer term experiments may further elucidate the seasonal dynamics and mechanism of Sphagnum allelopathy.

Predicted structural proteome of Sphagnum divinum and proteome-scale annotation.

Sphagnum-dominated peatlands store a substantial amount of terrestrial carbon. The genus is undersampled and under-studied. No experimental crystal structure from any Sphagnum species exists in the Protein Data Bank and fewer than 200 Sphagnum-related genes have structural models available in the AlphaFold Protein Structure Database. Tools and resources are needed to help bridge these gaps, and to enable the analysis of other structural proteomes now made possible by accurate structure prediction. We present the predicted structural proteome (25134 primary transcripts) of Sphagnum divinum computed using AlphaFold, structural alignment results of all high-confidence models against an annotated nonredundant crystallographic database of over 90,000 structures, a structure-based classification of putative Enzyme Commission (EC) numbers across this proteome, and the computational method to perform this proteome-scale structure-based annotation. All data and code are available in public repositories, detailed at https://github.com/BSDExabio/SAFA. The structural models of the S. divinum proteome have been deposited in the ModelArchive repository at https://modelarchive.org/doi/10.5452/ma-ornl-sphdiv.

Effects of Water-Ethanol Extracts from Four Sphagnum Species on Gene Expression of Selected Enzymes in Normal Human Dermal Fibroblasts and Their Antioxidant Properties.

Mosses (Bryophyta), particularly species of the genus Sphagnum, which have been used for centuries for the treatment of skin diseases and damage, are still not explored enough in terms of their use in cosmetics. The purpose of this study was to determine the antioxidant properties of water-ethanol extracts from four selected species of the genus Sphagnum (S. girgenshonii Russow, S. magellanicum Brid., S. palustre L., and S. squarrosum Crome) and their impact on the expression of genes encoding key enzymes for the functioning of the skin. In this study, the effects of Sphagnum extracts on the expression of genes encoding tyrosinase, collagenase, elastase, hyaluronidase and hyaluronic acid synthase in human dermal fibroblasts were determined for the first time in vitro. The extracts inhibited tyrosinase gene expression and showed antioxidant activity. The experiment showed an increase in the expression of some genes encoding collagenase (MMP1) or hyaluronidase (HYAL2, HYAL3 and HYAL4) and a decrease in the hyaluronan synthase (HAS1, HAS2 and HAS3) genes expression by the tested extracts. The obtained results suggest that using extracts from the tested Sphagnum species in anti-aging cosmetics does not seem beneficial. Further studies are needed to clarify their impact on the skin.

Divergent selection and climate adaptation fuel genomic differentiation between sister species of Sphagnum (peat moss).

New plant species can evolve through the reinforcement of reproductive isolation via local adaptation along habitat gradients. Peat mosses (Sphagnaceae) are an emerging model system for the study of evolutionary genomics and have well-documented niche differentiation among species. Recent molecular studies have demonstrated that the globally distributed species Sphagnum magellanicum is a complex of morphologically cryptic lineages that are phylogenetically and ecologically distinct. Here, we describe the architecture of genomic differentiation between two sister species in this complex known from eastern North America: the northern S. diabolicum and the largely southern S. magniae. We sampled plant populations from across a latitudinal gradient in eastern North America and performed whole genome and restriction-site associated DNA sequencing. These sequencing data were then analyzed computationally. Using sliding-window population genetic analyses we find that differentiation is concentrated within 'islands' of the genome spanning up to 400 kb that are characterized by elevated genetic divergence, suppressed recombination, reduced nucleotide diversity and increased rates of non-synonymous substitution. Sequence variants that are significantly associated with genetic structure and bioclimatic variables occur within genes that have functional enrichment for biological processes including abiotic stress response, photoperiodism and hormone-mediated signalling. Demographic modelling demonstrates that these two species diverged no more than 225 000 generations ago with secondary contact occurring where their ranges overlap. We suggest that this heterogeneity of genomic differentiation is a result of linked selection and reflects the role of local adaptation to contrasting climatic zones in driving speciation. This research provides insight into the process of speciation in a group of ecologically important plants and strengthens our predictive understanding of how plant populations will respond as Earth's climate rapidly changes.

Clonality, local population structure and gametophyte sex ratios in cryptic species of the Sphagnum magellanicum complex.

Sphagnum (peatmoss) comprises a moss (Bryophyta) clade with ~300-500 species. The genus has unparalleled ecological importance because Sphagnum-dominated peatlands store almost a third of the terrestrial carbon pool and peatmosses engineer the formation and microtopography of peatlands. Genomic resources for Sphagnum are being actively expanded, but many aspects of their biology are still poorly known. Among these are the degree to which Sphagnum species reproduce asexually, and the relative frequencies of male and female gametophytes in these haploid-dominant plants. We assess clonality and gametophyte sex ratios and test hypotheses about the local-scale distribution of clones and sexes in four North American species of the S. magellanicum complex. These four species are difficult to distinguish morphologically and are very closely related. We also assess microbial communities associated with Sphagnum host plant clones and sexes at two sites. Four hundred and five samples of the four species, representing 57 populations, were subjected to restriction site-associated DNA sequencing (RADseq). Analyses of population structure and clonality based on the molecular data utilized both phylogenetic and phenetic approaches. Multi-locus genotypes (genets) were identified using the RADseq data. Sexes of sampled ramets were determined using a molecular approach that utilized coverage of loci on the sex chromosomes after the method was validated using a sample of plants that expressed sex phenotypically. Sex ratios were estimated for each species, and populations within species. Difference in fitness between genets was estimated as the numbers of ramets each genet comprised. Degrees of clonality [numbers of genets/numbers of ramets (samples)] within species, among sites, and between gametophyte sexes were estimated. Sex ratios were estimated for each species, and populations within species. Sphagnum-associated microbial communities were assessed at two sites in relation to Sphagnum clonality and sex. All four species appear to engage in a mixture of sexual and asexual (clonal) reproduction. A single ramet represents most genets but two to eight ramets were dsumbers ansd text etected for some genets. Only one genet is represented by ramets in multiple populations; all other genets are restricted to a single population. Within populations ramets of individual genets are spatially clustered, suggesting limited dispersal even within peatlands. Sex ratios are male-biased in S. diabolicum but female-biased in the other three species, although significantly so only in S. divinum. Neither species nor males/females differ in levels of clonal propagation. At St Regis Lake (NY) and Franklin Bog (VT), microbial community composition is strongly differentiated between the sites, but differences between species, genets and sexes were not detected. Within S. divinum, however, female gametophytes harboured two to three times the number of microbial taxa as males. These four Sphagnum species all exhibit similar reproductive patterns that result from a mixture of sexual and asexual reproduction. The spatial patterns of clonally replicated ramets of genets suggest that these species fall between the so-called phalanx patterns, where genets abut one another but do not extensively mix because of limited ramet fragmentation, and the guerrilla patterns, where extensive genet fragmentation and dispersal result in greater mixing of different genets. Although sex ratios in bryophytes are most often female-biased, both male and female biases occur in this complex of closely related species. The association of far greater microbial diversity for female gametophytes in S. divinum, which has a female-biased sex ratio, suggests additional research to determine if levels of microbial diversity are consistently correlated with differing patterns of sex ratio biases.