The crane fly glycosylated triketide δ-lactone cornicinine elicits akinete differentiation of the cyanobiont in aquatic Azolla fern symbioses.

The restriction of plant-symbiont dinitrogen fixation by an insect semiochemical had not been previously described. Here we report on a glycosylated triketide δ-lactone from Nephrotoma cornicina crane flies, cornicinine, that causes chlorosis in the floating-fern symbioses from the genus Azolla. Only the glycosylated trans-A form of chemically synthesized cornicinine was active: 500 nM cornicinine in the growth medium turned all cyanobacterial filaments from Nostoc azollae inside the host leaf-cavities into akinetes typically secreting CTB-bacteriocins. Cornicinine further inhibited akinete germination in Azolla sporelings, precluding re-establishment of the symbiosis during sexual reproduction. It did not impact development of the plant Arabidopsis thaliana or several free-living cyanobacteria from the genera Anabaena or Nostoc but affected the fern host without cyanobiont. Fern-host mRNA sequencing from isolated leaf cavities confirmed high NH4-assimilation and proanthocyanidin biosynthesis in this trichome-rich tissue. After cornicinine treatment, it revealed activation of Cullin-RING ubiquitin-ligase-pathways, known to mediate metabolite signaling and plant elicitation consistent with the chlorosis phenotype, and increased JA-oxidase, sulfate transport and exosome formation. The work begins to uncover molecular mechanisms of cyanobiont differentiation in a seed-free plant symbiosis important for wetland ecology or circular crop-production today, that once caused massive CO2 draw-down during the Eocene geological past.

Effects of ultraviolet and photosynthetically active radiation on morphogenesis, antioxidants and photoprotective defense mechanism in a hot-spring cyanobacterium Nostoc sp. strain VKB02.

The continuous increase in global temperature and ultraviolet radiation (UVR) causes profound impacts on the growth and physiology of photosynthetic microorganisms. The hot-spring cyanobacteria have a wide range of mitigation mechanisms to cope up against current unsustainable environmental conditions. In the present investigation, we have explored the indispensable mitigation strategies of an isolated hot-spring cyanobacterium Nostoc sp. strain VKB02 under simulated ultraviolet (UV-A, UV-B) and photosynthetically active radiation (PAR). The adaptive morphological changes were more significantly observed under PAB (PAR, UV-A, and UV-B) exposure as compared to P and PA (PAR and UV-A) irradiations. PAB exposure also exhibited a marked decline in pigment composition and photosynthetic efficiency by multi-fold increment of free radicals. To counteract the oxidative stress, enzymatic and non-enzymatic antioxidants defense were significantly enhanced many folds under PAB exposure as compared to the control. In addition, the cyanobacterium has also produced shinorine as a strong free radicals scavenger and excellent UV absorber for effective photoprotection against UV radiation. Therefore, the hot-spring cyanobacterium Nostoc sp. strain VKB02 has unique defense strategies for survival under prolonged lethal UVR conditions. This study will help in the understanding of environment-induced defense strategies and production of highly value-added green photo-protectants for commercial applications.

Novel metabolic interactions and environmental conditions mediate the boreal peatmoss-cyanobacteria mutualism.

Interactions between Sphagnum (peat moss) and cyanobacteria play critical roles in terrestrial carbon and nitrogen cycling processes. Knowledge of the metabolites exchanged, the physiological processes involved, and the environmental conditions allowing the formation of symbiosis is important for a better understanding of the mechanisms underlying these interactions. In this study, we used a cross-feeding approach with spatially resolved metabolite profiling and metatranscriptomics to characterize the symbiosis between Sphagnum and Nostoc cyanobacteria. A pH gradient study revealed that the Sphagnum-Nostoc symbiosis was driven by pH, with mutualism occurring only at low pH. Metabolic cross-feeding studies along with spatially resolved matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) identified trehalose as the main carbohydrate source released by Sphagnum, which were depleted by Nostoc along with sulfur-containing choline-O-sulfate, taurine and sulfoacetate. In exchange, Nostoc increased exudation of purines and amino acids. Metatranscriptome analysis indicated that Sphagnum host defense was downregulated when in direct contact with the Nostoc symbiont, but not as a result of chemical contact alone. The observations in this study elucidated environmental, metabolic, and physiological underpinnings of the widespread plant-cyanobacterial symbioses with important implications for predicting carbon and nitrogen cycling in peatland ecosystems as well as the basis of general host-microbe interactions.

Exposure of cyanobacterium Nostoc sp. to the Mars-like stratosphere environment.

During the HH-19-2 flight mission of the Chinese Scientific Experimental System, dried Nostoc sp. cells were exposed to the stratosphere environment (32,508 m altitude) for 3 h and 22 min. The atmospheric pressure, temperature, relative humidity, and ionizing and non-ionizing radiation levels at that altitude are similar to those on the surface of Mars. Although analyses revealed decreased photosynthetic activity, a decline in autofluorescence, and damage to the cellular morphology in the flight-exposed sample, the death rate was low (28%). Physiological changes were not obvious after the exposure to the Mars-like vacuum conditions. The ground-exposed samples showed a similar trend to the flight-exposed samples, but the damage was relatively slight. RNA-sequencing data revealed a number of affected metabolic pathways: photosynthetic system and CO2 fixation function, activation of antioxidant systems, heat shock protein, DNA repair, and protein synthesis. Results suggest that Nostoc sp. has the potential to survive in a Mars-like environment and that it may be a suitable pioneer species to colonize Mars in the future in closed life-support systems (base) or in localities with relatively suitable conditions for life, such as localities with water available.

ß-N-Methylamino-L-Alanine (BMAA) Causes Severe Stress in Nostoc sp. PCC 7120 Cells under Diazotrophic Conditions: A Proteomic Study.

Non-proteinogenic neurotoxic amino acid ß-N-methylamino-L-alanine (BMAA) is synthesized by cyanobacteria, diatoms, and dinoflagellates, and is known to be a causative agent of human neurodegenerative diseases. Different phytoplankton organisms' ability to synthesize BMAA could indicate the importance of this molecule in the interactions between microalgae in nature. We were interested in the following: what kinds of mechanisms underline BMAA's action on cyanobacterial cells in different nitrogen supply conditions. Herein, we present a proteomic analysis of filamentous cyanobacteria Nostoc sp. PCC 7120 cells that underwent BMAA treatment in diazotrophic conditions. In diazotrophic growth conditions, to survive, cyanobacteria can use only biological nitrogen fixation to obtain nitrogen for life. Note that nitrogen fixation is an energy-consuming process. In total, 1567 different proteins of Nostoc sp. PCC 7120 were identified by using LC-MS/MS spectrometry. Among them, 123 proteins belonging to different functional categories were selected-due to their notable expression differences-for further functional analysis and discussion. The presented proteomic data evidences that BMAA treatment leads to very strong (up to 80%) downregulation of α (NifD) and ß (NifK) subunits of molybdenum-iron protein, which is known to be a part of nitrogenase. This enzyme is responsible for catalyzing nitrogen fixation. The genes nifD and nifK are under transcriptional control of a global nitrogen regulator NtcA. In this study, we have found that BMAA impacts in a total of 22 proteins that are under the control of NtcA. Moreover, BMAA downregulates 18 proteins that belong to photosystems I or II and light-harvesting complexes; BMAA treatment under diazotrophic conditions also downregulates five subunits of ATP synthase and enzyme NAD(P)H-quinone oxidoreductase. Therefore, we can conclude that the disbalance in energy and metabolite amounts leads to severe intracellular stress that induces the upregulation of stress-activated proteins, such as starvation-inducible DNA-binding protein, four SOS-response enzymes, and DNA repair enzymes, nine stress-response enzymes, and four proteases. The presented data provide new leads into the ecological impact of BMAA on microalgal communities that can be used in future investigations.

The cyanobacterial taxis protein HmpF regulates type IV pilus activity in response to light.

Motility is ubiquitous in prokaryotic organisms including the photosynthetic cyanobacteria where surface motility powered by type 4 pili (T4P) is common and facilitates phototaxis to seek out favorable light environments. In cyanobacteria, chemotaxis-like systems are known to regulate motility and phototaxis. The characterized phototaxis systems rely on methyl-accepting chemotaxis proteins containing bilin-binding GAF domains capable of directly sensing light, and the mechanism by which they regulate the T4P is largely undefined. In this study we demonstrate that cyanobacteria possess a second, GAF-independent, means of sensing light to regulate motility and provide insight into how a chemotaxis-like system regulates the T4P motors. A combination of genetic, cytological, and protein-protein interaction analyses, along with experiments using the proton ionophore carbonyl cyanide m-chlorophenyl hydrazine, indicate that the Hmp chemotaxis-like system of the model filamentous cyanobacterium Nostoc punctiforme is capable of sensing light indirectly, possibly via alterations in proton motive force, and modulates direct interaction between the cyanobacterial taxis protein HmpF, and Hfq, PilT1, and PilT2 to regulate the T4P motors. Given that the Hmp system is widely conserved in cyanobacteria, and the finding from this study that orthologs of HmpF and T4P proteins from the distantly related model unicellular cyanobacterium Synechocystis sp. strain PCC6803 interact in a similar manner to their N. punctiforme counterparts, it is likely that this represents a ubiquitous means of regulating motility in response to light in cyanobacteria.

Cyanobacterial inoculation as resource conserving options for improving the soil nutrient availability and growth of maize genotypes.

Harnessing the benefits of plant-microbe interactions towards better nutrient mobilization and plant growth is an important challenge for agriculturists globally. In our investigation, the focus was towards analyzing the soil-plant-environment interactions of cyanobacteria-based formulations (Anabaena-Nostoc consortium, BF1-4 and Anabaena-Trichoderma biofilm, An-Tr) as inoculants for ten maize genotypes (V1-V10). Field experimentation using seeds treated with the formulations illustrated a significant increase of 1.3- to 3.8-fold in C-N mobilizing enzyme activities in plants, along with more than five- to six-fold higher values of nitrogen fixation in rhizosphere soil samples. An increase of 22-30% in soil available nitrogen was also observed at flag leaf stage, and 13-16% higher values were also recorded in terms of cob yield of V6 with An-Tr biofilm inoculation. Savings of 30 kg N ha-1 season-1 was indicative of the reduced environmental pollution, due to the use of microbial options. The use of cyanobacterial formulations also enhanced the economic, environmental and energy use efficiency. This was reflected as 37-41% reduced costs lowered GHG emission by 58-68 CO2 equivalents and input energy requirement by 3651-4296 MJ, over the uninoculated control, on hectare basis. This investigation highlights the superior performance of these formulations, not only in terms of efficient C-N mobilization in maize, but also making maize cultivation a more profitable enterprise. Such interactions can be explored as resource-conserving options, for future evaluation across ecologies and locations, particularly in the global climate change scenario.

Cold adaptation in drylands: transcriptomic insights into cold-stressed Nostoc flagelliforme and characterization of a hypothetical gene with cold and nitrogen stress tolerance.

Environmental stressors, especially low temperature, are very common on the earth's dryland systems. Terrestrial cyanobacteria have evolved with cold adaptability in addition to extreme dryness and high irradiation resistance. The dryland soil surface-dwelling species, Nostoc flagelliforme, serves as a potential model organism to gain insights into cyanobacterial cold adaptation. In this study, we performed transcriptomic analysis of N. flagelliforme samples in response to low temperature. The results revealed that the biological processes, such as terpenoid biosynthetic process, oxidoreductase activity, carbohydrate metabolism, biosynthesis of secondary metabolites, lipid and nitrogen metabolism, were significantly and dynamically changed during the cold stress. It was noteworthy that the transcription of the denitrification pathway for ammonia accumulation was enhanced, implying an importance for nitrogen utilization in stress resistance. In addition, characterization of a cold-responsive hypothetical gene csrnf1 found that it could greatly improve the cold-resistant performance of cells when it was heterologously expressed in transgenic Nostoc sp. PCC 7120. It was also found that csrnf1 transgenic strain exhibited resistance to nitrogen-deficient environmental stress. Considering that dryland cyanobacteria have to cope with low temperature on infertile soils, this study would enrich our understanding on the importance of multifunction of the genes for environmental cold adaptation in drylands.

HetL, HetR and PatS form a reaction-diffusion system to control pattern formation in the cyanobacterium nostoc PCC 7120.

Local activation and long-range inhibition are mechanisms conserved in self-organizing systems leading to biological patterns. A number of them involve the production by the developing cell of an inhibitory morphogen, but how this cell becomes immune to self-inhibition is rather unknown. Under combined nitrogen starvation, the multicellular cyanobacterium Nostoc PCC 7120 develops nitrogen-fixing heterocysts with a pattern of one heterocyst every 10-12 vegetative cells. Cell differentiation is regulated by HetR which activates the synthesis of its own inhibitory morphogens, diffusion of which establishes the differentiation pattern. Here, we show that HetR interacts with HetL at the same interface as PatS, and that this interaction is necessary to suppress inhibition and to differentiate heterocysts. hetL expression is induced under nitrogen-starvation and is activated by HetR, suggesting that HetL provides immunity to the heterocyst. This protective mechanism might be conserved in other differentiating cyanobacteria as HetL homologues are spread across the phylum.

Cyanobacteria are the only bacteria on Earth able to draw their energy directly from the sun in the same way that plants do. In addition, some strains are able to 'fix' the nitrogen present in the atmosphere: they can extract this gas and transform it into nitrogen-based compounds necessary for life. However, both processes cannot happen in a given cell at the same time. A strain of cyanobacteria called Nostoc PCC 7120 can organise itself into long filaments of interconnected cells. Under certain conditions, one in every ten cells stops drawing its energy from the sun, and starts fixing atmospheric nitrogen instead. Exactly how the bacteria are able to 'count to ten' and organize themselves in such a precise pattern is still unclear. Cells can communicate and establish patterns by exchanging molecular signals that switch on and off certain cell programs. For instance, a protein called HetR turns on the genetic program that allows cyanobacteria to fix nitrogen; on the other hand, a signal known as PatS binds to HetR and turns it off. Cells starting to specialise in fixing nitrogen produce both HetR and PatS, with the latter diffusing in surrounding cells and preventing them from extracting nitrogen. However, it remained unclear how the nitrogen-fixing cell could ignore its own PatS signal and keep its HetR signal active. HetL ­ another protein produced by the future nitrogen-fixing cell ­ could potentially play this role, but how it acts was unknown. Here, Xu et al. show that HetL cannot diffuse from one cell to the other, and that it binds to HetR at the same place than PatS does. When both PatS and HetL are present, they compete to attach to HetR, which stops PatS from turning off HetR and deactivating the nitrogen-fixing program. Understanding how cyanobacteria fix nitrogen could help to develop new types of natural fertiliser. More generally, dissecting how these simple organisms can create patterns could help to grasp how patterning emerges in more complex creatures.

Identification of a hormogonium polysaccharide-specific gene set conserved in filamentous cyanobacteria.

Cyanobacteria comprise a phylum defined by the capacity for oxygenic photosynthesis. Members of this phylum are frequently motile as well. Strains that display gliding or twitching motility across semisolid surfaces are powered by a conserved type IV pilus system (T4P). Among the filamentous, heterocyst-forming cyanobacteria, motility is usually confined to specialized filaments known as hormogonia, and requires the deposition of an associated hormogonium polysaccharide (HPS). The genes involved in assembly and export of HPS are largely undefined, and it has been hypothesized that HPS exits the outer membrane via an atypical T4P-driven mechanism. Here, several novel hps loci, primarily encoding glycosyl transferases, are identified. Mutational analysis demonstrates that the majority of these genes are essential for both motility and production of HPS. Notably, most mutant strains accumulate wild-type cellular levels of the major pilin PilA, but not extracellular PilA, indicating dysregulation of the T4P motors, and, therefore, a regulatory interaction between HPS assembly and T4P activity. A co-occurrence analysis of Hps orthologs among cyanobacteria identified an extended set of putative Hps proteins comprising most components of a Wzx/Wzy-type polysaccharide synthesis and export system. This implies that HPS may be secreted through a more canonical pathway, rather than a T4P-mediated mechanism.

Evidence for a mutualistic relationship between the cyanobacteria Nostoc and fungi Aspergilli in different environments.

Symbiotic partnerships are widespread in nature and in industrial applications yet there are limited examples of laboratory communities. Therefore, using common photobionts and mycobionts similar to those in natural lichens, we create an artificial lichen-like symbiosis. While Aspergillus nidulans and Aspergillus niger could not obtain nutrients from the green algae, Chlorella, and Scenedesmus, the cyanobacteria Nostoc sp. PCC 6720 was able to support fungal growth and also elevated the accumulation of total biomass. The Nostoc-Aspergillus co-cultures grew on light and CO2 in an inorganic BG11 liquid medium without any external organic carbon and fungal mycelia were observed to peripherally contact with the Nostoc cells in liquid and on solid media at lower cell densities. Overall biomass levels were reduced after implementing physical barriers to indicate that physical contact between cyanobacteria and heterotrophic microbes may promote symbiotic growth. The synthetic Nostoc-Aspergillus nidulans co-cultures also exhibited robust growth and stability when cultivated in wastewater over days to weeks in a semi-continuous manner when compared with axenic cultivation of either species. These Nostoc-Aspergillus consortia reveal species-dependent and mutually beneficial design principles that can yield stable lichen-like co-cultures and provide insights into microbial communities that can facilitate sustainability studies and broader applications in the future. KEY POINTS: • Artificial lichen-like symbiosis was built with wild-type cyanobacteria and fungi. • Physical barriers decreased biomass production from artificial lichen co-cultures. • Artificial lichen adapted to grow and survive in wastewater for 5 weeks.

Endophytic Colonization of Rice (Oryza sativa L.) by the Symbiotic Strain Nostoc punctiforme PCC 73102.

Cyanobacteria are phototrophic microorganisms able to establish nitrogen-fixing symbiotic associations with representatives of all four of the major phylogenetic divisions of terrestrial plants. Despite increasing knowledge on the beneficial effects of cyanobacteria in rice fields, the information about the interaction between these microorganisms and rice at the molecular and structural levels is still limited. We have used the model nitrogen-fixing cyanobacterium Nostoc punctiforme to promote a long-term stable endophytic association with rice. Inoculation with this strain of hydroponic cultures of rice produces a fast adherence of the cyanobacterium to rice roots. At longer times, cyanobacterial growth in the proximity of the roots increased until reaching a plateau. This latter phase coincides with the intracellular colonization of the root epidermis and exodermis. Structural analysis of the roots revealed that the cyanobacterium use an apoplastic route to colonize the plant cells. Moreover, plant roots inoculated with N. punctiforme show both the presence of heterocysts and nitrogenase activity, resulting in the promotion of plant growth under nitrogen deficiency, thus providing benefits for the plant.

Dynamic expression of intra- and extra-cellular proteome and the influence of epiphytic bacteria for Nostoc flagelliforme in response to rehydration.

Nostoc flagelliforme is well known for its strong ecological adaptability in inhabiting desert biological soil crusts. However, the mechanism of its recovery from quiescent to active state after prolonged dormancy remains poorly characterized. Especially how exoproteome be related to the adaptive strategies and participate in the microalgae-bacteria interaction. In the present work, we analysed the intra- and extra-cellular proteome of N. flagelliforme over a complete rehydration period both in sterilization and in natural condition for the first time. The protein expression profile for N. flagelliforme has more fluctuations during the first 1 h after wetting but been relatively steady after fully hydrated. According to the extracellular proteomic datasets, we found a dynamic secretion of various extracellular hydrolytic enzymes and membrane transport proteins, which were related to peptidoglycan digestion and nutrient exchange respectively. Two-hundred and thirteen differentially expressed proteins induced by sterilization also reflect variation in nutrient exchange and highlight symbiosis between N. flagelliforme and surrounding bacteria. We also identified 112 phosphopeptides and 217 phosphorylation site of 95 protein of hydrated N. flagelliforme. The time course datasets we present here will be a reference for understanding the molecular processes underlying N. flagelliforme resuscitation and its potential role in microbial community diversification and soil desertification control.

The Integrity of the Cell Wall and Its Remodeling during Heterocyst Differentiation Are Regulated by Phylogenetically Conserved Small RNA Yfr1 in Nostoc sp. Strain PCC 7120.

Yfr1 is a strictly conserved small RNA in cyanobacteria. A bioinformatic prediction to identify possible interactions of Yfr1 with mRNAs was carried out by using the sequences of Yfr1 from several heterocyst-forming strains, including Nostoc sp. strain PCC 7120. The results of the prediction were enriched in genes encoding outer membrane proteins and enzymes related to peptidoglycan biosynthesis and turnover. Heterologous expression assays with Escherichia coli demonstrated direct interactions of Yfr1 with mRNAs of 11 of the candidate genes. The expression of 10 of them (alr2458, alr4550, murC, all4829, all2158, mraY, alr2269, alr0834, conR, patN) was repressed by interaction with Yfr1, whereas the expression of amiC2, encoding an amidase, was increased. The interactions between Yfr1 and the 11 mRNAs were confirmed by site-directed mutagenesis of Yfr1. Furthermore, a Nostoc strain with reduced levels of Yfr1 had larger amounts of mraY and murC mRNAs, supporting a role for Yfr1 in the regulation of those genes. Nostoc strains with either reduced or increased expression of Yfr1 showed anomalies in cell wall completion and were more sensitive to vancomycin than the wild-type strain. Furthermore, growth in the absence of combined nitrogen, which involves the differentiation of heterocysts, was compromised in the strain overexpressing Yfr1, and filaments were broken at the connections between vegetative cells and heterocysts. These results indicate that Yfr1 is an important regulator of cell wall homeostasis and correct cell wall remodeling during heterocyst differentiation.IMPORTANCE Bacterial small RNAs (sRNAs) are important players affecting the regulation of essentially every aspect of bacterial physiology. The cell wall is a highly dynamic structure that protects bacteria from their fluctuating environment. Cell envelope remodeling is particularly critical for bacteria that undergo differentiation processes, such as spore formation or differentiation of heterocysts. Heterocyst development involves the deposition of additional layers of glycolipids and polysaccharides outside the outer membrane. Here, we show that a cyanobacterial phylogenetically conserved small regulatory RNA, Yfr1, coordinates the expression of proteins involved in cell wall-related processes, including peptidoglycan metabolism and transport of different molecules, as well as expression of several proteins involved in heterocyst differentiation.

Physiological and thylakoid ultrastructural changes in cyanobacteria in response to toxic manganese concentrations.

In this study, two cyanobacterial strains (morphologically identified as Microcystis novacekii BA005 and Nostoc paludosum BA033) were exposed to different Mn concentrations: 7.0, 10.5, 15.7, 23.6 and 35.4 mg L-1 for BA005; and 15.0, 22.5, 33.7, 50.6, and 76.0 mg L-1 for BA033. Manganese toxicity was assessed by growth rate inhibition (EC50), chlorophyll a content, quantification of Mn accumulation in biomass and monitoring morphological and ultrastructural effects. The Mn EC50 values were 16 mg L-1 for BA005 and 39 mg L-1 for BA033, respectively. Reduction of chlorophyll a contents and ultrastructural changes were observed in cells exposed to Mn concentrations greater than 23.6 and 33.7 mg L-1 for BA005 and BA033. Damage to intrathylakoid spaces, increased amounts of polyphosphate granules and an increased number of carboxysomes were observed in both strains. In the context of the potential application of these strains in bioremediation approaches, BA005 was able to remove Mn almost completely from aqueous medium after 96 h exposure to an initial concentration of 10.5 mg L-1, and BA033 was capable of removing 38% when exposed to initial Mn concentration of 22.5 mg L-1. Our data shed light on how these cyanobacterial strains respond to Mn stress, as well as supporting their utility as organisms for monitoring Mn toxicity in industrial wastes and potential bioremediation application.

Dryland cyanobacterial exopolysaccharides show protection against acid deposition damage.

Soil surface-dwelling cyanobacteria constitute an important part of the dryland ecosystem. The exopolysaccharide (EPS) matrix they establish plays multiple roles in helping cells cope with harsh environments and also improves soil physicochemical properties. Anthropogenic atmospheric nitrogen or sulfur depositions have arisen as an important environmental change in drylands. The acid moisture derived from the depositions will be absorbed by cyanobacterial EPS matrix and thus may pose a threat to cells. In this communication, we evaluated this potential impact in a dryland cyanobacterium, Nostoc flagelliforme, which is a representative polysaccharide-rich species and shows remarkable resistance to desiccation stress. A strong and resilient pH buffering property was found for the EPS matrix, mainly of the polysaccharide's role, and this could protect the cells from acid damage of pH 4-6, a general acidity range of rainwater in the world. Unlike in acid aquatic environments, terrestrial xeric environments ensure N. flagelliforme unlikely to undertake lasting severe acidification. Thus, protection of the EPS matrix for dryland cyanobacteria would be conducive to sustain their growth and ecological roles in face of atmospheric acid pollution.

Orange and red carotenoid proteins are involved in the adaptation of the terrestrial cyanobacterium Nostoc flagelliforme to desiccation.

The remarkable drought-resistance of the terrestrial cyanobacterium Nostoc flagelliforme (N. flagelliforme) has attracted attention for many years. In this study, we purified a group of red proteins that accumulate in dried field samples of N. flagelliforme. These red proteins contain canthaxanthin as the bound chromophore. Native-PAGE analysis revealed that the purified red proteins resolved into six visible red bands and were composed of four helical carotenoid proteins (HCPs), HCP1, HCP2, HCP3, and HCP6 (homologs to the N-terminal domain of the orange carotenoid protein (OCP)). Seven genes encode homologs of the OCP in the genome of N. flagelliforme: two full-length ocp genes (ocpx1 and ocpx2), four N-terminal domain hcp genes (hcp1, hcp2, hcp3, and hcp6), and one C-terminal domain ccp gene. The expression levels of hcp1, hcp2, and hcp6 were highly dependent on the water status of field N. flagelliforme samples, being downregulated during rehydration and upregulated during subsequent dehydration. Transcripts of ocpx2 were dominant in the dried field samples, which we confirmed by detecting the presence of OCPx2-derived peptides in the purified red proteins. The results shed light on the relationship between carotenoid-binding proteins and the desiccation resistance of terrestrial cyanobacteria, and the physiological functions of carotenoid-binding protein complexes in relation to desiccation are discussed.

Functional sustainability of periphytic biofilms in organic matter and Cu2+ removal during prolonged exposure to TiO2 nanoparticles.

Responses of microbial communities to nanotoxicity in aquatic ecosystems are largely unknown, particularly with respect to relationship between community dynamics and functions. Here, periphytic biofilms were selected as a model of species-rich microbial communities to elucidate their responses when exposed to titanium dioxide nanoparticles (TiO2-NPs). Especially, the relationships between the functions (e.g. organic matter and Cu2+ removal) and community dynamics after long-term exposure to TiO2-NPs were assessed systematically. After 5days exposure to TiO2-NPs (5mgL-1), periphytic biofilms showed sustainable functions in pollutant removal and strong plasticity in defensing the toxic disturbance of TiO2-NPs, including photosynthesis and carbon metabolic diversity. The sustainable pollutant removal functions of periphytic biofilms were attributed to their functional redundancy. Specifically, periphytic biofilms altered their composition with cyanobacteria, Sphingobacteriia and Spirochaetes being the newly dominant taxa, and changed the carbon substrate utilization pattern to maintain high photosynthesis and metabolic rates. Moreover, extracellular polymeric substances (EPS) especially proteins were overproduced to bind the NPs and thereby reduce the nanotoxicity. The information obtained in this study may greatly help to understand the interactions between microbial community dynamics and function under NPs exposure conditions and functional redundancy is an important mechanism of periphytic biofilms to maintain sustainable functions.

Cyanobacterial Catalase Activity Prevents Oxidative Stress Induced by Pseudomonas fluorescens DUS1-27 from Inhibiting Brassica napus L. (canola) Growth.

Plant growth-promoting bacteria (PGPB) inhabit the rhizosphere of plants and are capable of enhancing plant growth through a number of mechanisms. A strain of Pseudomonas fluorescens DUS1-27 was identified as a potential PGPB candidate based on its ability to increase the growth of Brassica napus L. (canola) over that of uninoculated control plants in a soil-based system. The same P. fluorescens isolate was found to reduce plant growth in a hydroponic growth system, with plants showing the symptoms of a microbe-associated molecular pattern (MAMP) response to the bacteria. The amperometric quantification of H2O2, fluorescence-based total peroxidase assays, and quantification of catalase gene expression levels using qRT-PCR revealed that oxidative stress reduced plant growth in the hydroponic system. The addition of the cyanobacterium Nostoc punctiforme (known to have high catalase activity levels) in the hydroponic system as a co-inoculant reduced oxidative stress (49.7% decrease in H2O2 concentrations) triggered by the addition of P. fluorescens DUS1-27, thereby enabling plants to grow larger than uninoculated control plants. These results show the advantage of inoculating with multiple bacteria to promote plant growth and, for the first time, demonstrate that N. punctiforme beneficially assists plants under oxidative stress through its catalase activity in planta.

Distinctive characters of Nostoc genomes in cyanolichens.

BACKGROUND: Cyanobacteria of the genus Nostoc are capable of forming symbioses with a wide range of organism, including a diverse assemblage of cyanolichens. Only certain lineages of Nostoc appear to be able to form a close, stable symbiosis, raising the question whether symbiotic competence is determined by specific sets of genes and functionalities. RESULTS: We present the complete genome sequencing, annotation and analysis of two lichen Nostoc strains. Comparison with other Nostoc genomes allowed identification of genes potentially involved in symbioses with a broad range of partners including lichen mycobionts. The presence of additional genes necessary for symbiotic competence is likely reflected in larger genome sizes of symbiotic Nostoc strains. Some of the identified genes are presumably involved in the initial recognition and establishment of the symbiotic association, while others may confer advantage to cyanobionts during cohabitation with a mycobiont in the lichen symbiosis. CONCLUSIONS: Our study presents the first genome sequencing and genome-scale analysis of lichen-associated Nostoc strains. These data provide insight into the molecular nature of the cyanolichen symbiosis and pinpoint candidate genes for further studies aimed at deciphering the genetic mechanisms behind the symbiotic competence of Nostoc. Since many phylogenetic studies have shown that Nostoc is a polyphyletic group that includes several lineages, this work also provides an improved molecular basis for demarcation of a Nostoc clade with symbiotic competence.