The Role of Serine/Threonine-Specific Protein Kinases in Cyanobacteria - SpkB Is Involved in Acclimation to Fluctuating Conditions in Synechocystis sp. PCC 6803.

Protein phosphorylation via serine/threonine protein kinases (Spk) is a widespread mechanism to adjust cellular processes toward changing environmental conditions. To study their role(s) in cyanobacteria, we investigated a collection of 11 completely segregated spk mutants among the 12 annotated Spks in the model cyanobacterium Synechocystis sp. PCC 6803. Screening of the mutant collection revealed that especially the mutant defective in SpkB encoded by slr1697 showed clear deviations regarding carbon metabolism, that is, reduced growth rates at low CO2 or in the presence of glucose, and different glycogen accumulation patterns compared to WT. Alterations in the proteome of ΔspkB indicated changes of the cell surface but also metabolic functions. A phospho-proteome analysis revealed the absence of any phosphorylation in two proteins, while decreased phosphorylation of the carboxysome-associated protein CcmM and increased phosphorylation of the allophycocyanin alpha subunit ApcA was detected in ΔspkB. Furthermore, the regulatory PII protein appeared less phosphorylated in the mutant compared to WT, which was verified in Western blot experiments, indicating a clearly delayed PII phosphorylation in cells shifted from nitrate-containing to nitrate-free medium. Our results indicate that SpkB is an important regulator in Synechocystis that is involved in phosphorylation of the PII protein and additional proteins.

Photosynthetic response of Chara braunii towards different bicarbonate concentrations.

A variety of inorganic carbon acquisition modes have been proposed in Characean algae, however, a broadly applicable inorganic carbon uptake mechanism is unknown for the genus Chara. In the present study, we analyzed if C. braunii can efficiently use HCO3 - as a carbon source for photosynthesis. For this purpose, C. braunii was exposed to different concentrations of NaHCO3 - at different time scales. The photosynthetic electron transport through photosystem I (PSI) and II (PSII), the maximum electron transport rate (ETRmax ), the efficiency of the electron transport rate (α, the initial slope of the ETR), and the light saturation point of photosynthesis (Ek ) were evaluated. Additionally, pigment contents (chlorophyll a, chlorophyll b, and carotenoids) were determined. Bicarbonate addition positively affected ETRmax , after direct HCO3 - application, of both PSII and PSI, but this effect seems to decrease after 1 h and 24 h. Similar trends were seen for Ek , but no significant effect was observed for α. Pigment contents showed no significant changes in relation to different HCO3 - concentrations. To evaluate if cyclic electron flow around PSI was involved in active HCO3 - uptake, the ratio of PSI ETRmax /PSII ETRmax was calculated but did not show a distinctive trend. These results suggest that C. braunii can utilize NaHCO3 - in short-term periods as a carbon source but could rely on other carbon acquisition mechanisms over prolonged time periods. These observations suggest that the minor role of HCO3 - as a carbon source for photosynthesis in this alga might differentiate C. braunii from other examined Chara spp.

The high-light-induced protein SliP4 binds to NDH1 and photosystems facilitating cyclic electron transport and state transition in Synechocystis sp. PCC 6803.

An increasing number of small proteins has been identified in the genomes of well-annotated organisms, including the model cyanobacterium Synechocystis sp. PCC 6803. We describe a newly assigned protein comprising 37 amino acids that is encoded upstream of the superoxide dismutase SodB encoding gene. To clarify the role of SliP4, we analyzed a Synechocystis sliP4 mutant and a strain containing a fully active, Flag-tagged variant of SliP4 (SliP4.f). The initial hypothesis that this small protein might be functionally related to SodB could not be supported. Instead, we provide evidence that it fulfills important functions related to the organization of photosynthetic complexes. Therefore, we named it a small light-induced protein of 4 kDa, SliP4. This protein is strongly induced under high-light conditions. The lack of SliP4 causes a light-sensitive phenotype due to impaired cyclic electron flow and state transitions. Interestingly, SliP4.f was co-isolated with NDH1 complex and both photosystems. The interaction between SliP4.f and all three types of complexes was further confirmed by additional pulldowns and 2D-electrophoreses. We propose that the dimeric SliP4 serves as a molecular glue promoting the aggregation of thylakoid complexes, which contributes to different electron transfer modes and energy dissipation under stress conditions.

The impact of salt stress on the physiology and the transcriptome of the model streptophyte green alga Chara braunii.

Chara braunii is a model for early land plant evolution and terrestrialization. Salt stress has a profound effect on water and ion transport activities, thereby interacting with many other processes, including inorganic carbon acquisition for photosynthesis. In this study, we analyzed the impact of salt stress (5 practical salt units, PSU) on the physiology and gene expression in C. braunii. Photosynthesis was only slightly affected 6 h after salt addition and returned to control levels after 48 h. Several organic compounds such as proline, glutamate, sucrose, and 2-aminobutyrate accumulated in salt-treated thalli and might contribute to osmotic potential acclimation, whereas the amount of K+ decreased. We quantified transcript levels for 17,387 genes, of which 95 were up-regulated and 44 down-regulated after salt addition. Genes encoding proteins of the functional groups ion/solute transport and cell wall synthesis/modulation were enriched among the up-regulated genes 24-48 h after salt stress, indicating their role in osmotic acclimation. However, a homolog to land plant ERD4 osmosensors was transiently upregulated after 6 h, and phylogenetic analyses suggested that these sensors evolved in Charophyceae. Down-regulated genes were mainly related to photosynthesis and carbon metabolism/fixation, consistent with the observed lowered growth after extended cultivation. The changed expression of genes encoding proteins for inorganic carbon acquisition might be related to the impact of salt on ionic relations and inorganic carbon uptake. The results indicate that C. braunii can tolerate enhanced salt concentrations in a defined acclimation process, including distinct gene expression changes to achieve new metabolic homeostasis.

The cell walls of different Chara species are characterized by branched galactans rich in 3-O-methylgalactose and absence of AGPs.

Streptophyte algae are the closest relatives to land plants; their latest common ancestor performed the most drastic adaptation in plant evolution around 500 million years ago: the conquest of land. Besides other adaptations, this step required changes in cell wall composition. Current knowledge on the cell walls of streptophyte algae and especially on the presence of arabinogalactan-proteins (AGPs), important signalling molecules in all land plants, is limited. To get deeper insights into the cell walls of streptophyte algae, especially in Charophyceae, we performed sequential cell wall extractions of four Chara species. The three species Chara globularis, Chara subspinosa and Chara tomentosa revealed comparable cell wall compositions, with pectins, xylans and xyloglucans, whereas Chara aspera stood out with higher amounts of uronic acids in the pectic fractions and lack of reactivity with antibodies binding to xylan- and xyloglucan epitopes. Search for AGPs in the four Chara species and in Nitellopsis obtusa revealed the presence of galactans with pyranosidic galactose in 1,3-, 1,6- and 1,3,6-linkage, which are typical galactan motifs in land plant AGPs. A unique feature of these branched galactans was high portions of 3-O-methylgalactose. Only Nitellopsis contained substantial amounts of arabinose A bioinformatic search for prolyl-4-hydroxylases, involved in the biosynthesis of AGPs, revealed one possible functional sequence in the genome of Chara braunii, but no hydroxyproline could be detected in the four Chara species or in Nitellopsis obtusa. We conclude that AGPs that is typical for land plants are absent, at least in these members of the Charophyceae.

The seasonal effects on the encrustation of charophytes in two hard-water lakes.

Encrustation and element content of six charophyte species from two hard-water lakes were investigated monthly for a period of 1 year. Seasonal patterns were analyzed for the interaction of water chemistry. Encrustation followed a seasonal pattern for Chara contraria, Chara subspinosa, and Nitellopsis obtusa in Lake Krüselin and for Chara globularis and Chara tomentosa in Lake Lützlow. However, no seasonality in the precipitated CaCO3 was observed for C. subspinosa in Lake Lützlow and for C. tomentosa in Lake Krüselin, indicating a lake-specific dependency. Species-specific encrustation was found. Chara contraria and N. obtusa encrusted the most in June and August, whereas C. subspinosa and Nitella flexilis/opaca exhibited lowest encrustation in March and April. The precipitated CaCO3 of charophytes correlated negatively to the concentration of total inorganic carbon in both lakes. Element content of plant dry weight was species-specific for Ca and K, and lake-specific for Mg. No specific pattern was found for the TP and Fe contents. The results showed seasonal, species, and lake-specific influences on the encrustation of charophytes.

Consequence of altered nitrogen cycles in the coupled human and ecological system under changing climate: The need for long-term and site-based research.

Anthropogenically derived nitrogen (N) has a central role in global environmental changes, including climate change, biodiversity loss, air pollution, greenhouse gas emission, water pollution, as well as food production and human health. Current understanding of the biogeochemical processes that govern the N cycle in coupled human-ecological systems around the globe is drawn largely from the long-term ecological monitoring and experimental studies. Here, we review spatial and temporal patterns and trends in reactive N emissions, and the interactions between N and other important elements that dictate their delivery from terrestrial to aquatic ecosystems, and the impacts of N on biodiversity and human society. Integrated international and long-term collaborative studies covering research gaps will reduce uncertainties and promote further understanding of the nitrogen cycle in various ecosystems.

The gene sml0013 of Synechocystis species strain PCC 6803 encodes for a novel subunit of the NAD(P)H oxidoreductase or complex I that is ubiquitously distributed among Cyanobacteria.

The NAD(P)H oxidoreductase or complex I (NDH1) complex participates in many processes such as respiration, cyclic electron flow, and inorganic carbon concentration in the cyanobacterial cell. Despite immense progress in our understanding of the structure-function relation of the cyanobacterial NDH1 complex, the subunits catalyzing NAD(P)H docking and oxidation are still missing. The gene sml0013 of Synechocystis 6803 encodes for a small protein of unknown function for which homologs exist in all completely known cyanobacterial genomes. The protein exhibits weak similarities to the NDH-dependent flow6 (NDF6) protein, which was reported from Arabidopsis (Arabidopsis thaliana) chloroplasts as a NDH subunit. An sml0013 inactivation mutant of Synechocystis 6803 was generated and characterized. It showed only weak differences regarding growth and pigmentation in various culture conditions; most remarkably, it exhibited a glucose-sensitive phenotype in the light. The genome-wide expression pattern of the Δsml0013::Km mutant was almost identical to the wild type when grown under high CO2 conditions as well as after shifts to low CO2 conditions. However, measurements of the photosystem I redox kinetic in cells of the Δsml0013::Km mutant revealed differences, such as a decreased capability of cyclic electron flow as well as electron flow into respiration in comparison with the wild type. These results suggest that the Sml0013 protein (named NdhP) represents a novel subunit of the cyanobacterial NDH1 complex, mediating its coupling either to the respiratory or the photosynthetic electron flow.

Wide ecological niches ensure frequent harmful dinoflagellate blooms.

Harmful algal blooms (HABs) and their consequences cause multiple devastating effects in various freshwater, brackish and marine ecosystems. However, HAB species at moderate population densities have positive ecological roles as primary producers of organic matter and food for zooplankton and fish. They also enhance benthic-pelagic coupling and participate in the biogeochemical cycles. The consequences of HABs are transported across the conventional environmental boundaries by numerous cascade effects in the food webs and beyond. Meanwhile, forecasts of bloom events are still limited, largely because of scarcity of reliable information on ecological niches of the bloom-forming algae. To fill up this knowledge gap, this study focused on dinoflagellates, a diverse group of mostly photosynthesizing protists (unicellular eukaryotes) capable of mixotrophy, since they play a key role in primary production and formation of blooms in marine and brackish waters worldwide. In this study, ecological niches of 17 abundant bloom-forming dinoflagellate species from coastal regions of the southern Baltic Sea were identified for the first time. It was hypothesized that wider ecological niches ensure more frequent dinoflagellate blooms compared to the species with narrower niches. This hypothesis was verified using the long-term (44 years) database on phytoplankton abundance and physical-chemical characteristics of the environment. It were analyzed 4534 datasets collected from 1972 to 2016. Fourteen abiotic parameters (water temperature, salinity, Secchi depth, pH, Chl a, and concentration of basic nutrients) were considered as ecological niche dimensions. The Principal Component Analysis presented the dissolved inorganic nitrogen, total nitrogen, Chl a, and temperature as principal niche dimensions of dinoflagellates. The algal bloom criteria were refined. It was for the first time proved statistically that HAB frequency of dinoflagellate species robustly correlated with the width of their ecological niches.

From sink to source: Dynamic of greenhouse gases emissions from beach wrack accumulations in a temperate coastal bay.

Coastal ecosystems such as salt marshes, seagrass meadows, and kelp forests contribute to climate regulation as carbon sinks. However, coastal ecosystems may act as carbon sources as beach wrack accumulations may release greenhouse gases (GHG) during decomposition. The magnitude of GHG emissions of beach wrack accumulations under natural conditions are poorly understood, hampering accurate blue carbon accountings. In this study, we assessed the spatio-temporal variability and environmental factors driving CO2, CH4 and N2O emissions from beach wrack accumulations on a temperate sandy beach. Beach wrack accumulations, dominated by Zostera marina and opportunistic brown macroalgae, presented variable spatio-temporal dynamics. Annual beach wrack GHG emissions achieved up to 77,915 mg m-2 d-1 CO2e (CO2 equivalents) and varied largely throughout the study period due to interactive effects of temperature, wave exposure, beach wrack biomass moisture, abundance, and species composition. Our findings showed that methane emissions in new, freshly deposited, and in drifting wrack in the water reached up to 100 mg m-2 d-1, representing up to 57 % of annual CO2e emissions occurring throughout the year. Nitrous oxide emissions were highly variable and comprised a minor extent (i.e., up to 4 %) of annual CO2e emissions. Together, wrack CH4 and N2O emissions provided 13.69 g CO2 m-2 per year to the atmosphere. Our findings indicate that excessive opportunistic macroalgae biomass driven by eutrophication may explain increased CO2 and N2O emissions. We conclude that whilst beach wrack depositions are a natural and essential part of coastal ecosystems, they may provide an extra source of GHG to the atmosphere, potentially counteracting the role of vegetated coastal ecosystems as carbon sinks.

Plastid DNA sequences and oospore characters of some European taxa of Tolypella section Tolypella (Characeae) identify five clusters, including one new cryptic Tolypella taxon from Sardinia, but they do not coincide with current morphological descriptions.

In Europe, the genus Tolypella (Characeae) comprises four to eight Tolypella taxa in sections Rothia and Tolypella that have been distinguished by vegetative morphology and gametangial characters such as antheridial size and oospore wall ornamentation. However, morphological differentiation is difficult in some cases due to overlapping and variable vegetative features, which in many cases are difficult to observe clearly. To clarify the taxonomic status of the five European taxa of Tolypella in section Tolypella, sequence data of the plastid genes atpB, rbcL and psbC for Tolypella glomerata (Desv.) Leonh., Tolypella hispanica Allen, Tolypella nidifica (O.F. Müll.) A. Braun, Tolypella normaniana (Nordst.) Nordst. and Tolypella salina Cor. were combined with data on oospore morphology, including oospore wall ornamentation. Gene sequence data identified five distinct clusters, but they were not consistent with the morphologically identified five taxa. T. glomerata consisted of some of the samples morphologically identified as T. glomerata and seven samples of T. normaniana, while the remaining T. glomerata samples clustered with specimens of unclear affiliation (Tolypella sp.). We identified two clusters of T. hispanica within the European material: cluster T. hispanica I consisted of samples from various locations, whereas the second cluster (T. hispanica II) consisted of samples of T. hispanica from Sardinia Island. The remaining cluster consisted of all the specimens that had been determined as T. salina or T. nidifica in addition to two specimens of T. normaniana. Oospore morphology was most clearly distinguishable for T. glomerata. Oospore characteristics for all other taxa were not as informative but showed some geographical and/or environmentally influenced differences, especially for T. nidifica and T. salina. Our results suggest the need to further check the different taxonomy of Tolypella sect. Tolypella in which specimens normally identified as T. glomerata might be two different taxa, T. glomerata and an unidentified taxon; T. nidifica and T. salina are not separate taxa; T. normaniana is a diminutive variant of two different Tolypella taxa; and T. hispanica comprises two different taxa, one from the Mediterranean island Sardinia.

Two parthenogenetic populations of Chara canescens differ in their capacity to acclimate to irradiance and salinity.

The parthenogens of Chara canescens (Charophyceae) occupy broader geographical and ecological ranges than their sexual counterparts. Two possible hypotheses explain the ubiquity of parthenogens: the occurrence of one or several parthenogens with wide niches, or of many parthenogens that are restricted to narrow ecological niches. For the purposes of this study, C. canescens individuals from two neighbouring populations of the Baltic Sea (Bodstedter Bodden = BB; Salzhaff = SH), which differed significantly in water transparency and salinity, were investigated for significant differences in physiological capacity. Individuals of both habitats acclimated quickly to daily changes in irradiances in the field, but the photosynthetic efficiency of PS II showed a significant decrease with increasing daily irradiance in the habitat BB, which has lower levels of salinity and water transparency. In addition to the field study, individuals were reared under different levels of environmental factors in the laboratory: four irradiances (70-600 µmol m(-2) s(-1)) and five salinity levels (0-24 psu). The individuals of both habitats grew almost equally well at intermediate salinity levels. Growth under the artificial light supply was highest at levels corresponding to the in situ conditions for each population. Total chlorophyll was highest at intermediate salinities (BB), or hardly changed with salinity (SH). The physiological capacity for individuals from SH clearly depends upon changing growth irradiance, whereas the capacity for individuals from BB was relatively independent of salinity and irradiance. These findings indicate that both parthenogenetic C. canescens populations are locally adapted to light. However, to test adaptive potential of the parthenogens, more than two populations should be tested in future.

Establishment and optimization of a new model organism to study early land plant evolution: Germination, cultivation and oospore variation of Chara braunii Gmelin, 1826.

For studying land plant evolution, the establishment and optimization of model organisms representing streptophytic algae, sister to land plants, is essential. Long-term cultivation experiments with Chara braunii S276 were performed over 8 years, since 4 years (Nov. 2018) under constant conditions. Additionally, short-term experiments for optimization of culture conditions were performed with three strains of C. braunii (S276, NIES-1604 and Lausiger Teiche, LaT-2708). Germination success after application of sterilization agents, addition of gibberellic acid and under different incubation conditions with respect to pre-treatment, irradiance regime and substrate was investigated in order to develop protocols for generative cultivation of at least unialgal cultures. The resulting cultivation protocols for C. braunii S276, allowing maintenance of vegetative as well as generative cultures are presented in detail, including protocols for germination induction and growth of sterilized and unsterilized oospores.

Stress ecology in fucus: abiotic, biotic and genetic interactions.

Stress regimes defined as the synchronous or sequential action of abiotic and biotic stresses determine the performance and distribution of species. The natural patterns of stress to which species are more or less well adapted have recently started to shift and alter under the influence of global change. This was the motivation to review our knowledge on the stress ecology of a benthic key player, the macroalgal genus Fucus. We first provide a comprehensive review of the genus as an ecological model including what is currently known about the major lineages of Fucus species with respect to hybridization, ecotypic differentiation and speciation; as well as life history, population structure and geographic distribution. We then review our current understanding of both extrinsic (abiotic/biotic) and intrinsic (genetic) stress(es) on Fucus species and how they interact with each other. It is concluded that (i) interactive stress effects appear to be equally distributed over additive, antagonistic and synergistic categories at the level of single experiments, but are predominantly additive when averaged over all studies in a meta-analysis of 41 experiments; (ii) juvenile and adult responses to stress frequently differ and (iii) several species or particular populations of Fucus may be relatively unaffected by climate change as a consequence of pre-adapted ecotypes that collectively express wide physiological tolerences. Future research on Fucus should (i) include additional species, (ii) include marginal populations as models for responses to environmental stress; (iii) assess a wider range of stress combinations, including their temporal fluctuations; (iv) better differentiate between stress sensitivity of juvenile versus adult stages; (v) include a functional genomic component in order to better integrate Fucus' ecological and evolutionary responses to stress regimes and (vi) utilize a multivariate modelling approach in order to develop and understand interaction networks.

Chara zeylanica J.G.Klein ex Willd. (Charophyceae, Charales, Characeae): First European Record from the Island of Sardinia, Italy.

The first record of a species belonging to the genus Chara L. subgenus Chara R.D.Wood section Grovesia R.D.Wood subsect. Willdenowia R.D.Wood from Europe is presented here, thus challenging the interpretation of its distribution pattern as an intertropical group of charophytes. The morphological characters of the specimens, as well as the results of a phylogenetic analysis, clearly identified them as Chara zeylanica J.G.Klein ex Willd. Although the subsection Willdenowia has yet to receive a thorough taxonomic treatment, a discussion of its relationship to other taxa of this subsection is provided despite the lack of a commonly agreed upon taxonomic concept. The ecological conditions of the Sardinian site of C. zeylanica are presented. Moreover, the status of and threats to this taxon, and hypotheses regarding potential pathways through which it reached Europe, are discussed.

Bioremediation approach using charophytes-preliminary laboratory and field studies of mine drainage water from the Mansfeld Region, Germany.

Mine drainage water from the Schlenze stream, Mansfeld Region, Central Germany, which have shown an increase in heavy metal concentrations of Cd2+, Cu2+, Pb2+, and Zn2+, was used to investigate the bioremediation potential of charophytes. The removal of heavy metals by Chara subspinosa from the water was tested in single- and multi-metal additions. The uptake capacity of C. subspinosa decreased during the course of the experiment and was higher in single-metal addition than in multi-metal addition of Pb2+, Zn2+, and Cd2+. Accumulation of heavy metals in the carbonate encrustation of charophytes was far lower than those to which they were exposed. Cu, Cd, Pb, and Zn co-precipitated more in the encrustation of C. subspinosa exposed to single-metal approach than to multi-metal approach. The carbonate composition of charophytes was influenced by the water chemistry. Content of Na in the carbonate encrustation correlated with the Na+ concentration of the respective water. The toxic effect of heavy metals on photosynthesis was species-specific. Electron transport rates (ETRmax) were higher in Chara tomentosa than in C. subspinosa. Charophytes withstand the heavy metal concentrations when diluted with river water from the Altarm cut-off lake and can therefore be used for the bioremediation of diluted mine drainage waters by co-precipitating Cd, Cu, and Zn.

Chaos theory discloses triggers and drivers of plankton dynamics in stable environment.

Despite the enticing discoveries of chaos in nature, triggers and drivers of this phenomenon remain a classical enigma which needs irrefutable empirical evidence. Here we analyze results of the yearlong replicated mesocosm experiment with multi-species plankton community that allowed revealing signs of chaos at different trophic levels in strictly controlled abiotic environment. In mesocosms without external stressors, we observed the "paradox of chaos" when biotic interactions (internal drivers) were acting as generators of internal abiotic triggers of complex plankton dynamics. Chaos was registered as episodes that vanished unpredictably or were substituted by complex behaviour of other candidates when longer time series were considered. Remarkably, episodes of chaos were detected even in the most abiotically stable conditions. We developed the Integral Chaos Indicator to validate the results of the Lyapunov exponent analysis. These findings are essential for modelling and forecasting behaviour of a variety of natural and other global systems.

Geographic variation in fitness-related traits of the bladderwrack Fucus vesiculosus along the Baltic Sea-North Sea salinity gradient.

In the course of the ongoing global intensification and diversification of human pressures, the study of variation patterns of biological traits along environmental gradients can provide relevant information on the performance of species under shifting conditions. The pronounced salinity gradient, co-occurrence of multiple stressors, and accelerated rates of change make the Baltic Sea and its transition to North Sea a suitable region for this type of study. Focusing on the bladderwrack Fucus vesiculosus, one of the main foundation species on hard-bottoms of the Baltic Sea, we analyzed the phenotypic variation among populations occurring along 2,000 km of coasts subjected to salinities from 4 to >30 and a variety of other stressors. Morphological and biochemical traits, including palatability for grazers, were recorded at 20 stations along the Baltic Sea and four stations in the North Sea. We evaluated in a common modeling framework the relative contribution of multiple environmental drivers to the observed trait patterns. Salinity was the main and, in some cases, the only environmental driver of the geographic trait variation in F. vesiculosus. The decrease in salinity from North Sea to Baltic Sea stations was accompanied by a decline in thallus size, photosynthetic pigments, and energy storage compounds, and affected the interaction of the alga with herbivores and epibiota. For some traits, drivers that vary locally such as wave exposure, light availability or nutrient enrichment were also important. The strong genetic population structure in this macroalgae might play a role in the generation and maintenance of phenotypic patterns across geographic scales. In light of our results, the desalination process projected for the Baltic Sea could have detrimental impacts on F. vesiculosus in areas close to its tolerance limit, affecting ecosystem functions such as habitat formation, primary production, and food supply.

Age and site-specific pattern on encrustation of charophytes.

Encrustation and element content (Ca, Fe, K, Mg and P) of charophytes was studied along plant thalli to investigate the dependency of thallus age and site-specificity. Charophytes were collected from five sampling sites (Angersdorfer Teiche, Asche, Bruchwiesen, Krüselinsee and Lützlower See) which were distinct with respect to water chemistry. Furthermore, photosynthesis was measured to identify the physiological state of plants in habitat waters and with the addition of different ion concentrations (Ca2+, K+, Mg2+ and Na+). Age pattern on encrustation of charophytes was site-specific: carbonate content increased from the youngest to the oldest part (Angersdorfer Teiche), younger parts were less encrusted than older parts in Asche, Bruchwiesen and Krüselinsee, whereas encrustation in Lützlower See was the same along plants thallus. Charophytes showed species-specific encrustation in investigated sites. Encrustation of C. hispida in Angersdorfer Teiche was also as high as of individuals from hard-water lakes irrespective of 10.15 mS cm-1 (salinity of 6.3). For species growing in Angersdorfer Teiche, K/Na content and photosynthesis was lowest when compared to other sites. Photosynthesis of charophytes was enhanced after the addition of KCl and adversely affected by CaCl2, MgCl2 and NaCl. In summary, it was shown that encrustation of charophytes in water sites with strong ion anomalies could be as high as in hard-water lakes. It is assumed that ion composition, rather than ion concentration of Na+, Mg2+ and SO42-, impact on the encrustation of charophytes. The age pattern on encrustation in this study showed a strong site-specificity, whereas encrustation of charophytes was species-specific. Ion concentrations, either of habitats or actively added in laboratory measurements, impact on encrustation, element content and photosynthesis of charophytes.

Eelgrass Leaf Surface Microbiomes Are Locally Variable and Highly Correlated with Epibiotic Eukaryotes.

Eelgrass (Zostera marina) is a marine foundation species essential for coastal ecosystem services around the northern hemisphere. Like all macroscopic organisms, it possesses a microbiome (here defined as an associated prokaryotic community) which may play critical roles in modulating the interaction of eelgrass with its environment. For example, its leaf surface microbiome could inhibit or attract eukaryotic epibionts which may overgrow the eelgrass leading to reduced primary productivity and subsequent eelgrass meadow decline. We used amplicon sequencing of the 16S and 18S rRNA genes of prokaryotes and eukaryotes to assess the leaf surface microbiome (prokaryotes) as well as eukaryotic epibionts in- and outside lagoons on the German Baltic Sea coast. Prokaryote microbiomes varied substantially both between sites inside lagoons and between open coastal and lagoon sites. Water depth, leaf area and biofilm chlorophyll a concentration explained a large amount of variation in both prokaryotic and eukaryotic community composition. The prokaryotic microbiome and eukaryotic epibiont communities were highly correlated, and network analysis revealed disproportionate co-occurrence between a limited number of eukaryotic taxa and several bacterial taxa. This suggests that eelgrass leaf surfaces are home to a mosaic of microbiomes of several epibiotic eukaryotes, in addition to the microbiome of the eelgrass itself. Our findings thereby underline that eukaryotic diversity should be taken into account in order to explain prokaryotic microbiome assembly and dynamics in aquatic environments.