Production Of Compatible Solutes Research Articles

Psychromarinibacter sediminicola sp. nov., a novel moderately halophilic, metabolically diverse bacterium isolated from a solar saltern sediment, and comparison between members of family Roseobacteraceae.

Known for its species abundance and evolutionary status complexity, family Roseobacteraceae is an important subject of many studies on the discovery, identification, taxonomic status, and ecological properties of marine bacteria. This study compared and analyzed the phylogenetic, genomic, biochemical, and chemo taxonomical properties of seven species from three genera (Psychromarinibacter, Lutimaribacter, and Maritimibacter) of the family Roseobacteraceae. Moreover, a novel strain, named C21-152T was isolated from solar saltern sediment in Weihai, China. The values of 16S rRNA gene sequence similarity, the average nucleotide identity (ANI), the average amino acid identity (AAI), and the digital DNA-DNA hybridization (dDDH) between genomes of the novel strain and Psychromarinibacter halotolerans MCCC 1K03203T were 97.19, 78.49, 73.45, and 21.90%, respectively. Genome sequencing of strain C21-152T revealed a complete Sox enzyme system related to thiosulfate oxidization as well as a complete pathway for the final conversion of hydroxyproline to α-ketoglutarate. In addition, strain C21-152T was resistant to many antibiotics and had the ability to survive below 13% salinity. This strain had versatile survival strategies in saline environments including salt-in, compatible solute production and compatible solute transport. Some of its physiological features enriched and complemented the knowledge of the characteristics of the genus Psychromarinibacter. Optimum growth of strain C21-152T occurred at 37℃, with 5-6% (w/v) NaCl and at pH 7.5. According to the results of the phenotypic, chemotaxonomic characterization, phylogenetic properties and genome analysis, strain C21-152T should represent a novel specie of the genus Psychromarinibacter, for which the name Psychromarinibacter sediminicola sp. nov. is proposed. The type strain is C21-152T (= MCCC 1H00808T = KCTC 92746T = SDUM1063002T).

Simulating compatible solute biosynthesis using a metabolic flux model of the biomining acidophile, Acidithiobacillus ferrooxidans ATCC 23270

Halotolerant, acidophilic, bioleaching microorganisms are crucial to biomining operations that utilize saline water. Compatible solutes play an important role in the adaptation of these microorganisms to saline environments. Acidithiobacillus ferrooxidans ATCC 23270, an iron- and sulfur-oxidizing acidophilic bacterium, synthesizes trehalose as its native compatible solute but is still sensitive to salinity. Recently, halotolerant bioleaching bacteria were found to use ectoine as their key compatible solute. Previously, bioleaching bacteria were recalcitrant to genetic manipulation; however, recent advancements in genetic tools and techniques allow successful genetic modification of A. ferrooxidans ATCC 23270. Therefore, this study aimed to test, in silico, the effect of native and synthetic compatible solute biosynthesis by A. ferrooxidans ATCC 23270 on its growth and metabolism. Metabolic network flux modelling was used to provide a computational framework for the prediction of metabolic fluxes during production of native and synthetic compatible solutes by A. ferrooxidans ATCC 23270, in silico. Complete pathways for trehalose biosynthesis by the bacterium are proposed and captured in the updated metabolic model including a newly discovered UDP-dependent trehalose synthesis pathway. Finally, the effect of nitrogen sources on compatible solute production was simulated and showed that using nitrogen gas as the sole nitrogen source enables the ectoine-producing ‘engineered’ microbe to oxidize up to 20% more ferrous iron in comparison to the native microbe that only produces trehalose. Therefore, the predictive outcomes of the model have the potential to guide the design and optimization of a halotolerant strain of A. ferrooxidans ATCC 23270 for saline bioleaching operations.

Salinity alleviator bacteria in rice (Oryza sativa L.), their colonization efficacy, and synergism with melatonin.

In this study, rhizospheric and endophytic bacteria were tested for the alleviation of salinity stress in rice. Endophytic isolates were taken from previous studies based on their salt stress-alleviating traits. The rhizospheric bacteria were isolated from rice and screened based on salt tolerance and plant growth-promoting traits. Molecular identification indicated the presence of class Gammaproteobacteria, Bacillota, and Actinomycetia. Two-two most potential isolates each from rhizospheric and endophytic bacteria were selected for in planta trials. Results showed that microbial inoculation significantly improved germination and seedling vigor under elevated salinity. The confocal scanning laser microscopy showed higher bacterial colonization in inoculated rice roots than in control. Based on this experiment, rhizospheric bacteria Brevibacterium frigoritolerans W19 and endophytic Bacillus safensis BTL5 were selected for pot trial along with a growth-inducing compound melatonin 20 ppm. Inoculation of these two bacteria improved the levels of chlorophyll, proline, phenylalanine ammonia-lyase, catalase, superoxide dismutase, polyphenol oxidase, root-shoot length, and dry weight under elevated salt concentration. The gene expression studies showed modulation of SOD1, CATa, NHX1, and PAL1 genes by the bacterial strains and melatonin application. The inoculation was found to have additive effects with 20 ppm melatonin. This enhancement in dry matter accumulation, compatible solute production, and oxidative stress regulation could help plants in mitigating the ill effects of high salinity. Exploring such a combination of microbes and inducer molecules could be potentially useful in developing stress-alleviating bioformulations.

Functional Insights of Salinity Stress-Related Pathways in Metagenome-Resolved Methanothrix Genomes.

ABSTRACTRecently, methanogenic archaea belonging to the genus Methanothrix were reported to have a fundamental role in maintaining stable ecosystem functioning in anaerobic bioreactors under different configurations/conditions. In this study, we reconstructed three Methanothrix metagenome-assembled genomes (MAGs) from granular sludge collected from saline upflow anaerobic sludge blanket (UASB) reactors, where Methanothrix harundinacea was previously implicated with the formation of compact and stable granules under elevated salinity levels (up to 20 g/L Na+). Genome annotation and pathway analysis of the Methanothrix MAGs revealed a genetic repertoire supporting their growth under high salinity. Specifically, the most dominant Methanothrix (MAG_279), classified as a subspecies of Methanothrix_A harundinacea_D, had the potential to augment its salinity resistance through the production of different glycoconjugates via the N-glycosylation process, and via the production of compatible solutes as Nε-acetyl-β-lysine and ectoine. The stabilization and reinforcement of the cell membrane via the production of isoprenoids was identified as an additional stress-related pathway in this microorganism. The improved understanding of the salinity stress-related mechanisms of M. harundinacea highlights its ecological niche in extreme conditions, opening new perspectives for high-efficiency methanisation of organic waste at high salinities, as well as the possible persistence of this methanogen in highly-saline natural anaerobic environments.IMPORTANCE Using genome-centric metagenomics, we discovered a new Methanothrix harundinacea subspecies that appears to be a halotolerant acetoclastic methanogen with the flexibility for adaptation in the anaerobic digestion process both at low (5 g/L Na+) and high salinity conditions (20 g/L Na+). Annotation of the recovered M. harundinacea genome revealed salinity stress-related functions, including the modification of EPS glycoconjugates and the production of compatible solutes. This is the first study reporting these genomic features within a Methanothrix sp., a milestone further supporting previous studies that identified M. harundinacea as a key-driver in anaerobic granulation under high salinity stress.

Impact of Two Strains of Rhizobium leguminosarum on the Adaptation to Terminal Water Deficit of Two Cultivars Vicia faba.

Drought stress has become one of the most uncontrolled and unpredictable constraints on crop production. The purpose of this study was to evaluate the impacts of two different Rhizobium leguminosarum strains on terminal drought tolerance induction in two faba bean genotypes cultivated in Algeria, Aquadulce and Maltais. To this end, we measured physiological parameters—osmoprotectants accumulation, oxidative stress markers and enzyme activities—to assess the effect of R. leguminosarum inoculation on V. faba under terminal water deficiency conditions in greenhouse trials. Upregulation of anti-oxidative mechanisms and production of compatible solutes were found differentially activated according to Rhizobium strain. Drought stress resilience of the Maltais variety was improved using the local Rhizobium strain OL13 compared to the common strain 3841. Symbiosis with OL13 strain leads in particular to a much better production of proline and soluble sugar in nodules but also in roots and leaves of Maltais plant. Even if additional work is still necessary to decipher the mechanism by which a Rhizobium strain can affect the accumulation of osmoprotectants or cellular redox status in all the plants, inoculation with selected Rhizobium could be a promising strategy for improving water stress management in the forthcoming era of climate change.

Heterologous Production of Glycine Betaine Using Synechocystis sp. PCC 6803-Based Chassis Lacking Native Compatible Solutes.

Among compatible solutes, glycine betaine has various applications in the fields of nutrition, pharmaceuticals, and cosmetics. Currently, this compound can be extracted from sugar beet plants or obtained by chemical synthesis, resulting in low yields or high carbon footprint, respectively. Hence, in this work we aimed at exploring the production of glycine betaine using the unicellular cyanobacterium Synechocystis sp. PCC 6803 as a photoautotrophic chassis. Synechocystis mutants lacking the native compatible solutes sucrose or/and glucosylglycerol—∆sps, ∆ggpS, and ∆sps∆ggpS—were generated and characterized. Under salt stress conditions, the growth was impaired and accumulation of glycogen decreased by ∼50% whereas the production of compatible solutes and extracellular polymeric substances (capsular and released ones) increased with salinity. These mutants were used as chassis for the implementation of a synthetic device based on the metabolic pathway described for the halophilic cyanobacterium Aphanothece halophytica for the production of the compatible solute glycine betaine. Transcription of ORFs comprising the device was shown to be stable and insulated from Synechocystis’ native regulatory network. Production of glycine betaine was achieved in all chassis tested, and was shown to increase with salinity. The introduction of the glycine betaine synthetic device into the ∆ggpS background improved its growth and enabled survival under 5% NaCl, which was not observed in the absence of the device. The maximum glycine betaine production [64.29 µmol/gDW (1.89 µmol/mg protein)] was reached in the ∆ggpS chassis grown under 3% NaCl. Taking into consideration this production under seawater-like salinity, and the identification of main key players involved in the carbon fluxes, this work paves the way for a feasible production of this, or other compatible solutes, using optimized Synechocystis chassis in a pilot-scale.

How salt stress-responsive proteins regulate plant adaptation to saline conditions.

An overview is presented of recent advances in our knowledge of candidate proteins that regulate various physiological and biochemical processes underpinning plant adaptation to saline conditions. Salt stress is one of the environmental constraints that restrict plant distribution, growth and yield in many parts of the world. Increased world population surely elevates food demands all over the globe, which anticipates to add a great challenge to humanity. These concerns have necessitated the scientists to understand and unmask the puzzle of plant salt tolerance mechanisms in order to utilize various strategies to develop salt tolerant crop plants. Salt tolerance is a complex trait involving alterations in physiological, biochemical, and molecular processes. These alterations are a result of genomic and proteomic complement readjustments that lead to tolerance mechanisms. Proteomics is a crucial molecular tool that indicates proteins expressed by the genome, and also identifies the functions of proteins accumulated in response to salt stress. Recently, proteomic studies have shed more light on a range of promising candidate proteins that regulate various processes rendering salt tolerance to plants. These proteins have been shown to be involved in photosynthesis and energy metabolism, ion homeostasis, gene transcription and protein biosynthesis, compatible solute production, hormone modulation, cell wall structure modification, cellular detoxification, membrane stabilization, and signal transduction. These candidate salt responsive proteins can be therefore used in biotechnological approaches to improve tolerance of crop plants to salt conditions. In this review, we provided comprehensive updated information on the proteomic data of plants/genotypes contrasting in salt tolerance in response to salt stress. The roles of salt responsive proteins that are potential determinants for plant salt adaptation are discussed. The relationship between changes in proteome composition and abundance, and alterations observed in physiological and biochemical features associated with salt tolerance are also addressed.

The Ecophysiological Performance and Traits of Genera within the Stichococcus-like Clade (Trebouxiophyceae) under Matric and Osmotic Stress.

Changes in water balance are some of the most critical challenges that aeroterrestrial algae face. They have a wide variety of mechanisms to protect against osmotic stress, including, but not limited to, downregulating photosynthesis, the production of compatible solutes, spore and akinete formation, biofilms, as well as triggering structural cellular changes. In comparison, algae living in saline environments must cope with ionic stress, which has similar effects on the physiology as desiccation in addition to sodium and chloride ion toxicity. These environmental challenges define ecological niches for both specialist and generalist algae. One alga known to be aeroterrestrial and euryhaline is Stichococcus bacillaris Nägeli, possessing the ability to withstand both matric and osmotic stresses, which may contribute to wide distribution worldwide. Following taxonomic revision of Stichococcus into seven lineages, we here examined their physiological responses to osmotic and matric stress through a salt growth challenge and desiccation experiment. The results demonstrate that innate compatible solute production capacity under salt stress and desiccation tolerance are independent of one another, and that salt tolerance is more variable than desiccation tolerance in the Stichococcus-like genera. Furthermore, algae within this group likely occupy similar ecological niches, with the exception of Pseudostichococcus.

Fascinating regulatory mechanism of silicon for alleviating drought stress in plants

Plants confront several environmental stresses that are crucial in defining plant productivity. Among these environmental stresses, drought stress is recognized as the foremost abiotic factor which affects the food security around the globe due to its negative impact on the plant development, and quality of plant products. Because of this, drought stress has gained an imperative appearance in the field of plant sciences in recent years. Silicon (Si), an agronomically essential mineral nutrient, is recognized highly advantageous in enhancing plant growth at various phases of plant life cycle under water deficit circumstances. Si has been considered extensively useful in mitigating harmful consequences of drought stress by enhancing root H2O absorption, regulating uptake of nutrients, reducing transpiration rate, improving photosynthetic activity, increment in the production of compatible solutes and by elevating plant antioxidant defense functioning. Si also known to improve plant endurance to limited water availability by inducing the functioning of various stress associated genes. To maximize the potential benefits and sustainable efficiency of Si in agriculture, it is of crucial importance to gain knowledge about the underlying mechanisms of how Si counteract stress conditions. Various findings suggested that Si increases the plant's immune system against drought stress, and application of Si is an important approach to shield plants from adverse stress conditions and soil nutrient depletion. This paper shows that Si has imperative and noteworthy impacts on improving plant tolerance to drought stress via maintaining cellular homeostasis. Present review mainly provides an insight into silicon role as beneficial element to alleviate drought stress in plants by regulating their morphological, physicochemical and molecular characteristics.

Contact-dependent traits in Pseudomonas syringae B728a.

Production of the biosurfactant syringafactin by the plant pathogen Pseudomonas syringae B728a is a surface contact-dependent trait. Expression of syfA, as measured using a gfp reporter gene fusion was low in planktonic cells in liquid cultures but over 4-fold higher in cells immobilized on surfaces as varied as glass, plastic, paper, parafilm, agar, membrane filters, and leaves. Induction of syfA as measured by GFP fluorescence was rapid, occurring within two hours after immobilization of cells on surfaces. Comparison of the global transcriptome by RNA sequencing of planktonic cells in a nutrient medium with that of cells immobilized for 2 hours on filters placed on this solidified medium revealed that, in addition to syfA, 3156 other genes were differentially expressed. Genes repressed in immobilized cells included those involved in quaternary ammonium compound (QAC) metabolism and transport, compatible solute production, carbohydrate metabolism and transport, organic acid metabolism and transport, phytotoxin synthesis and transport, amino acid metabolism and transport, and secondary metabolism. Genes induced in immobilized cells included syfA plus those involved in translation, siderophore synthesis and transport, nucleotide metabolism and transport, flagellar synthesis and motility, lipopolysaccharide (LPS) synthesis and transport, energy generation, transcription, chemosensing and chemotaxis, replication and DNA repair, iron-sulfur proteins, peptidoglycan/cell wall polymers, terpenoid backbone synthesis, iron metabolism and transport, and cell division. That many genes are rapidly differentially expressed upon transfer of cells from a planktonic to an immobilized state suggests that cells experience the two environments differently. It seems possible that surface contact initiates anticipatory changes in P. syringae gene expression, which enables rapid and appropriate physiological responses to the different environmental conditions such as might occur in a biofilm. Such responses could help cells survive transitions from aquatic habitats fostering planktonic traits to attachment on surfaces, conditions that alternatively occur on leaves.

Identification and Expression Analysis of Salinity-induced Genes in Rangpur lime (Citrus limonia)

Among the economically important horticultural crops, citrus is one of the most vulnerable crops to soil salinity. Rangpur lime is more tolerant to high soil salinity than other commonly used citrus rootstocks. However, the molecular mechanism involved in salinity tolerance has not been explored in Rangpur lime. In this study, a cDNA library was constructed from leaves of Rangpur lime watered for 30 days with 100 mmol · L−1 NaCl in tap water or tap water using suppression subtractive hybridization (SSH). Two hundred cDNA clones randomly selected from this library were sequenced, and an average of 569 bp was obtained from the majority of clones. Fifty-six salinity-induced genes, showing 2- to 6-fold increases in their expression levels, were identified by macroarray hybridizations. Salinity-induced genes were associated with transcription (5.36%), stress response and signaling (21.43%), metabolism (16.07%), transport facilitation (10.71%), photosynthesis (10.71%), protein synthesis and fate (19.64%) and cellular biogenesis (3.57%). Stress response- and signaling-related genes constituted the largest functional group, associated with the production of compatible solutes, regulation of stomatal movement, lipid modification, oxidative stress, antioxidant defense, and stress signaling. Expression levels of 13 identified genes were induced 1.8- to 3.1-fold, which were validated in salt-treated and untreated Rangpur lime. The functions of salinity-induced, stress-related genes and their potential roles in salinity tolerance in Rangpur lime were discussed.

Proteome Comparison Between Natural Desiccation-Tolerant Plants and Drought-Protected Caspicum annuum Plants by Microbacterium sp. 3J1.

Desiccation-tolerant plants are able to survive for extended periods of time in the absence of water. The molecular understanding of the mechanisms used by these plants to resist droughts can be of great value for improving drought tolerance in crops. This understanding is especially relevant in an environment that tends to increase the number and intensity of droughts. The combination of certain microorganisms with drought-sensitive plants can improve their tolerance to water scarcity. One of these bacteria is Microbacterium sp. 3J1, an actinobacteria able to protect pepper plants from drought. In this study, we supplemented drought-tolerant and drought-sensitive plant rhizospheres with Microbacterium sp. 3J1 and analyzed their proteomes under drought to investigate the plant-microbe interaction. We also compare this root proteome with the proteome found in desiccation-tolerant plants. In addition, we studied the proteome of Microbacterium sp. 3J1 subjected to drought to analyze its contribution to the plant-microbe interaction. We describe those mechanisms shared by desiccation-tolerant plants and sensitive plants protected by microorganisms focusing on protection against oxidative stress, and production of compatible solutes, plant hormones, and other more specific proteins.

Rhizobacteria Isolated from Saline Soil Induce Systemic Tolerance in Wheat (Triticum aestivum L.) against Salinity Stress

Halo-tolerant plant growth-promoting rhizobacteria (PGPR) have the inherent potential to cope up with salinity. Thus, they can be used as an effective strategy in enhancing the productivity of saline agro-systems. In this study, a total of 50 isolates were screened from the rhizospheric soil of plants growing in the salt range of Pakistan. Out of these, four isolates were selected based on their salinity tolerance and plant growth promotion characters. These isolates (SR1. SR2, SR3, and SR4) were identified as Bacillus sp. (KF719179), Azospirillum brasilense (KJ194586), Azospirillum lipoferum (KJ434039), and Pseudomonas stutzeri (KJ685889) by 16S rDNA gene sequence analysis. In vitro, these strains, in alone and in a consortium, showed better production of compatible solute and phytohormones, including indole acetic acid (IAA), gibberellic acid (GA), cytokinin (CK), and abscisic acid (ABA), in culture conditions under salt stress. When tested for inoculation, the consortium of all four strains showed the best results in terms of improved plant biomass and relative water content. Consortium-inoculated wheat plants showed tolerance by reduced electrolyte leakage and increased production of chlorophyll a, b, and total chlorophyll, and osmolytes, including soluble sugar, proline, amino acids, and antioxidant enzymes (superoxide dismutase, catalase, peroxidase), upon exposure to salinity stress (150 mM NaCl). In conclusion, plant growth-promoting bacteria, isolated from salt-affected regions, have strong potential to mitigate the deleterious effects of salt stress in wheat crop, when inoculated. Therefore, this consortium can be used as potent inoculants for wheat crop under prevailing stress conditions.

Comparative metabolomic responses of low- and high-cadmium accumulating genotypes reveal the cadmium adaptive mechanism in Brassica napus

Recently, oilseed rape has gathered interest for its ability to withstand elevated metal contents in plant, a key feature for remediation of contaminated soils. In this study, comparative and functional metabolomic analyses using liquid chromatography/mass spectrometry were undertaken to explore the metabolic basis of this attribute under cadmium (Cd) stress. Results revealed both conserved and differential metabolomic responses between genotype CB671 (tolerant Cd-accumulating) and its sensitive counterpart ZD622. CB671 responded to Cd stress by rearranging carbon flux towards production of compatible solutes, sugar storage forms and ascorbate, as well as jasmonates, ethylene and vitamin B6. Intriguingly, IAA abundance was reduced by 1.91-fold, which was in connection with tryptophan funnelling into serotonin (3.48-fold rise). In ZD622 by contrast, Cd provoked drastic depletion of carbohydrates and vitamins, but subtle hormones alteration. A striking accumulation of unsaturated fatty acids and oxylipins in CB671, paralleled by glycerophospholipids build-up and induction of inositol-derived signalling metabolites (up to 5.41-fold) suggested ability for prompt triggering of detoxifying mechanisms. Concomitantly, phytosteroids, monoterpenes and carotenoids were induced, denoting fine-tuned mechanisms for membrane maintenance, which was not evident in ZD622. Further, ZD622 markedly accumulated phenolics from upstream sub-classes of flavonoids; in CB671 however, a distinct phenolic wiring was activated, prioritizing anthocyanins and lignans instead. Along with cell wall (CW) saccharides, the activation of lignans evoked CW priming in CB671. Current results have demonstrated existence of notable metabolomic-based strategies for Cd tolerance in metal-accumulating oilseed rapes, and provided a holistic view of metabolites potentially contributing to Cd tolerance in this species.

Evidence for chaotropicity/kosmotropicity offset in a yeast growth model.

Chaotropes are compounds which cause the disordering, unfolding and denaturation of biological macromolecules. It is the chaotropicity of fermentation products that often acts as the primary limiting factor in ethanol and butanol fermentations. Since ethanol is mildly chaotropic at low concentrations, it prevents the growth of the producing microbes via its impacts on a variety of macromolecular systems and their functions. Kosmotropes have the opposite effect to chaotropes and we hypothesised that it might be possible to use these to mitigate chaotrope-induced inhibition of Saccharomyces cerevisiae growth. We also postulated that kosmotrope-mediated mitigation of chaotropicity is not quantitatively predictable. The chaotropes ethanol and urea, and compatible solutes glycerol and betaine (kosmotrope), and the highly kosmotropic salt ammonium sulphate all inhibited the growth rate of Saccharomyces cerevisiae in the concentration range 5-15%. They resulted in increased lag times, decreased maximum specific growth rates, and decreased final optical densities. Surprisingly, neither the stress protectants nor ammonium sulphate reduced the inhibition of growth caused by ethanol. Whereas, in some cases, compatible solutes and kosmotropes mitigated against the inhibitory effects of urea. However, this effect was not mathematically additive from the quantification of chao-/kosmotropicity of each individual compound. The potential effects of glycerol, betaine and/or ammonium sulphate may have been reduced or masked by the metabolic production of compatible solutes. It may nevertheless be that the addition of kosmotropes to fermentations which produce chaotropic products can enhance metabolic activity, growth rate, and/or product formation.

Compatible solutes profiling and carbohydrate feedstock from diversified cyanobacteria

Cyanobacteria are promising cell factories for producing high-value bioproducts. They accumulate low molecular mass organic molecules under salinity stress conditions, which are known as compatible solutes, e.g. sucrose, trehalose, and glucosylglycerol (GG). The physiological properties of compatible solutes sort them as the potential bioproducts for applications in the field of health, pharmacy, and cosmetic sectors. During last few decades, mostly model cyanobacterial strains (unicellular from freshwater and marine habitats) have been targeted for the biochemical and molecular characterization of compatible solutes accumulation, while, the study on other cyanobacteria from extreme/stressful habitats are scanty. The advantage for selecting multicellular cyanobacteria in the present study is their easier harvesting mode over unicellular strains which are one of the primary concerns for the cost-effective production of desired products at pilot scale. Therefore, five cyanobacterial strains with diverse morphotypes belonging to subsection I (Unicellular), II (Baeocytous), III (Filamentous), and V (Ramified) isolated from extreme/stressful habitats (sewage pond and hot spring including freshwater strain as a reference) were screened for salt tolerance, compatible solutes profiling, and sugar feedstock production. High GG, sucrose, and glycogen production were found in Leptolyngbya sp. PCC 8913, Chroogloeocystis siderophila NIES-1031 and Fischerella major NIES-592, respectively. Under salt stress, Leptolyngbya sp. PCC 8913 showed a GG production of 101 ± 12 mg/L/OD730, which is significantly higher than that of the model strain Synechocystis sp. PCC 6803. Fischerella major NIES-592 and Oscillatoria laetevirens NIES-31 were identified as novel trehalose-producing cyanobacteria. The key genes involved in production of compatible solutes among these cyanobacteria were also studied. Thus, the results not only displayed the potential cyanobacteria for production of GG, sucrose, trehalose and glycogen but also address the metabolic pathways for the compatible solutes biosynthesis in diversified cyanobacteria, which can be further targeted for the in-depth study to enhance the production using molecular approaches.

Effect of Seed Inoculation on Alfalfa Tolerance to Water Deficit Stress

Water deficit is one of the most important environmental stresses that adversely affect crop growth and production and mycorrhizal fungi and symbiotic bacteria have important role in resistance to drought stress. The effect of biofertilizers on alfalfa stress tolerance was studied at the greenhouse condition. Treatments comprised three water-deficit stresses (35%, 55% and 75% of field capacity) and four seeds inoculations (Glomus mosseae, Sinorhizobium meliloti, G. mosseae + S. meliloti and non-inoculated). Water-deficit stress decrease cell membrane stability (39%), total Chl (24.05%), carotenoid (35.55%), quantum yield (50.64%) and forage yield (28.20%), while increased the proline and soluble sugars content (68.55 and 46.53% respectively) and osmotic potential (45.84%). The inoculation of seeds increased the capability of the plants in counteracting the stress, so that the production of compatible solutes was increased and the photosynthetic indices, proline, osmotic potential, membrane stability and forage yield were improved by seed inoculation. Mycorrhiza improved photosynthetic indexes and proline, but bacteria had more efficacy on membrane stability and forage yield. However, double inoculation due to the synergistic effect of mycorrhiza and Sinorhizobium, had the greatest effect than Solitary inoculation. Our results suggest that biofertilized alfalfa plants were better adapted than non- biofertilized ones to cope with water deficit.

Belowground Response to Drought in a Tropical Forest Soil. I. Changes in Microbial Functional Potential and Metabolism.

Global climate models predict a future of increased severity of drought in many tropical forests. Soil microbes are central to the balance of these systems as sources or sinks of atmospheric carbon (C), yet how they respond metabolically to drought is not well-understood. We simulated drought in the typically aseasonal Luquillo Experimental Forest, Puerto Rico, by intercepting precipitation falling through the forest canopy. This approach reduced soil moisture by 13% and water potential by 0.14 MPa (from -0.2 to -0.34). Previous results from this experiment have demonstrated that the diversity and composition of these soil microbial communities are sensitive to even small changes in soil water. Here, we show prolonged drought significantly alters the functional potential of the community and provokes a clear osmotic stress response, including the production of compatible solutes that increase intracellular C demand. Subsequently, a microbial population emerges with a greater capacity for extracellular enzyme production targeting macromolecular carbon. Significantly, some of these drought-induced functional shifts in the soil microbiota are attenuated by prior exposure to a short-term drought suggesting that acclimation may occur despite a lack of longer-term drought history.

Hydrogen isotope response to changing salinity and rainfall in Australian mangroves.

Hydrogen isotope ratios ((2) H/(1) H, δ(2) H) of leaf waxes covary with those in precipitation and are therefore a useful paleohydrologic proxy. Mangroves are an exception to this relationship because their δ(2) H values are also influenced by salinity. The mechanisms underlying this response were investigated by measuring leaf lipid δ(2) H and leaf and xylem water δ(2) H and δ(18) O values from three mangrove species over 9.5 months in a subtropical Australian estuary. Net (2) H/(1) H fractionation between surface water and leaf lipids decreased by 0.5-1.0‰ ppt(-1) for n-alkanes and 0.4-0.8‰ ppt(-1) for isoprenoids. Xylem water was (2) H depleted relative to surface water, reflecting (2) H discrimination of 4-10‰ during water uptake at all salinities and opportunistic uptake of freshwater at high salinity. However, leaf water (2) H enrichment relative to estuary water was insensitive to salinity and identical for all species. Therefore, variations in leaf and xylem water δ(2) H values cannot explain the salinity-dependent (2) H depletion in leaf lipids, nor the 30‰ range in leaf lipid δ(2) H values among species. Biochemical changes in direct response to salt stress, such as increased compatible solute production or preferential use of stored carbohydrates, and/or the timing of lipid production and subsequent turnover rates, are more likely causes.

Metagenomic analysis reveals adaptations to a cold-adapted lifestyle in a low-temperature acid mine drainage stream.

An acid mine drainage (pH 2.5-2.7) stream biofilm situated 250 m below ground in the low-temperature (6-10°C) Kristineberg mine, northern Sweden, contained a microbial community equipped for growth at low temperature and acidic pH. Metagenomic sequencing of the biofilm and planktonic fractions identified the most abundant microorganism to be similar to the psychrotolerant acidophile, Acidithiobacillus ferrivorans. In addition, metagenome contigs were most similar to other Acidithiobacillus species, an Acidobacteria-like species, and a Gallionellaceae-like species. Analyses of the metagenomes indicated functional characteristics previously characterized as related to growth at low temperature including cold-shock proteins, several pathways for the production of compatible solutes and an anti-freeze protein. In addition, genes were predicted to encode functions related to pH homeostasis and metal resistance related to growth in the acidic metal-containing mine water. Metagenome analyses identified microorganisms capable of nitrogen fixation and exhibiting a primarily autotrophic lifestyle driven by the oxidation of the ferrous iron and inorganic sulfur compounds contained in the sulfidic mine waters. The study identified a low diversity of abundant microorganisms adapted to a low-temperature acidic environment as well as identifying some of the strategies the microorganisms employ to grow in this extreme environment.