Conditions Of Nitrogen Deprivation Research Articles

CPK28-mediated phosphorylation enhances nitrate transport activity of NRT2.1 during nitrogen deprivation.

Nitrate (NO3 -) serves as the primary inorganic nitrogen source assimilated by most terrestrial plants. The acquisition of nitrate from the soil is facilitated by NITRATE TRANSPORTERS (NRTs), with NRT2.1 being the key high-affinity nitrate transporter. The activity of NRT2.1, which has multiple potential phosphorylation sites, is intricately regulated under various physiological conditions. Here, we discovered that CALCIUM-DEPENDENT PROTEIN KINASE 28 (CPK28) positively regulates nitrate uptake under nitrogen deprivation conditions. We found CPK28 as the kinase targeted by immunoprecipitation followed by mass spectrometry and examined the in-planta phosphorylation status of NRT2.1 in cpk28 mutant plants by employing quantitative MS-based phosphoproteomics. Through a combination of in vitro phosphorylation experiment and immunoblotting using phospho-specific antibody, we successfully demonstrated that CPK28 specifically phosphorylates NRT2.1 at Ser21. Functional analysis conducted in Xenopus oocytes revealed that co-expression of CPK28 significantly enhanced high-affinity nitrate uptake of NRT2.1. Further investigation using transgenic plants showed that the phosphomimic variant NRT2.1S21E, but not the nonphosphorylatable variant NRT2.1S21A, fully restored high-affinity 15NO3 - uptake ability in both nrt2.1 and cpk28 mutant backgrounds. This study clarifies that the kinase activity of CPK28 is promoted during nitrogen deprivation conditions. These significant findings provide valuable insights into the intricate regulatory mechanisms that govern nitrate-demand adaptation.

Integration of Epigenome and Lactylome Reveals the Regulation of Lipid Production in Nannochloropsis oceanica.

Lysine lactylation (Kla) is a kind of novel post-translational modification (PTM) that participates in gene expression and various metabolic processes. Nannochloropsis has a remarkable capacity for triacylglycerol (TAG) production under nitrogen stress. To elucidate the involvement of lactylation in lipid synthesis, we conducted chromatin immunoprecipitation sequencing (ChIP-seq) and mRNA-seq analyses to monitor lactylation modifications and transcriptome alterations in Nannochloropsis oceanica. In all, 2057 genes showed considerable variation between nitrogen deprivation (ND) and nitrogen repletion (NR) conditions. Moreover, a total of 5375 differential Kla peaks were identified, including 5331 gain peaks and 44 loss peaks under ND vs NR. The differential Kla peaks were primarily distributed in the promoter (≤1 kb) (71.07%), 5'UTR (22.64%), and exon (4.25%). Integrative analysis of ChIP-seq, transcriptome, and previous proteome and lactylome data elucidates the potential mechanism by which lactylation promotes lipid accumulation under ND. Lactylation facilitates autophagy and protein degradation, leading to the recycling of carbon into the tricarboxylic acid (TCA) cycle, thereby providing carbon precursors for lipid synthesis. Additionally, lactylation induces the redirection of carbon from membrane lipids to TAG by upregulating lipases and enhancing the TCA cycle and β-oxidation pathways. This research offers a new perspective for the investigation of lipid biosynthesis in Nannochloropsis.

The cumulative impact of temperature and nitrogen availability on the potential nitrogen fixation and extracellular polymeric substances secretion by Dolichospermum

Nitrogen-fixing cyanobacteria not only cause severe blooms but also play an important role in the nitrogen input processes of lakes. The production of extracellular polymeric substances (EPS) and the ability to fix nitrogen from the atmosphere provide nitrogen-fixing cyanobacteria with a competitive advantage over other organisms. Temperature and nitrogen availability are key environmental factors in regulating the growth of cyanobacteria. In this study, Dolichospermum (formerly known as Anabaena) was cultivated at three different temperatures (10 °C, 20 °C, and 30 °C) to examine the impact of temperature and nitrogen availability on nitrogen fixation capacity and the release of EPS. Initially, confocal laser scanning microscopy (CLSM) and the quantification of heterocysts at different temperatures revealed that lower temperatures (10 °C) hindered the differentiation of heterocysts under nitrogen-deprived conditions. Additionally, while heterocysts inhibited the photosynthetic activity of Dolichospermum, the secretion of EPS was notably affected by nitrogen limitation, particularly at 30 °C. Finally, real-time quantitative polymerase chain reaction (qPCR) was used to measure the expression of nitrogen-utilizing genes (ntcA and nifH) and EPS synthesis-related genes (wzb and wzc). The results indicated that under nitrogen-deprived conditions, the expression of each gene was upregulated, and there was a significant correlation between the upregulation of nitrogen-utilizing and EPS synthesis genes (P < 0.05). Our findings suggested that Dolichospermum responded to temperature variation by affecting the formation of heterocysts, impacting its potential nitrogen fixation capacity. Furthermore, the quantity of EPS released was more influenced by nitrogen availability than temperature. This research enhances our comprehension of interconnections between nitrogen deprivation and EPS production under the different temperatures.

The fundamental role of pH in CH4 bioconversion into polyhydroxybutyrate in mixed methanotrophic cultures

Climate change and plastic pollution are likely the most relevant challenges for the environment in the 21st century. Developing cost-effective technologies for the bioconversion of methane (CH4) into polyhydroxyalkanoates (PHAs) could simultaneously mitigate CH4 emissions and boost the commercialization of biodegradable polymers. Despite the fact that the role of temperature, nitrogen deprivation, CH4:O2 ratio or micronutrients availability on the PHA accumulation capacity of methanotrophs has been carefully explored, there is still a need for optimization of the CH4-to-PHA bioconversion process prior to becoming a feasible platform in future biorefineries. In this study, the influence of different cultivation broth pH values (5.5, 7, 8.5 and 10) on bacterial biomass growth, CH4 bioconversion rate, PHA accumulation capacity and bacterial community structure was investigated in a stirred tank bioreactor under nitrogen deprivation conditions. Higher CH4 elimination rates were obtained at increasing pH, with a maximum value of 50.4 ± 2.7 g CH4·m−3·h−1 observed at pH 8.5. This was likely mediated by an increased ionic strength in the mineral medium, which enhanced the gas-liquid mass transfer. Interestingly, higher PHB accumulations were observed at decreasing pH, with the highest PHB contents recorded at a pH 5.5 (43.7 ± 3.4 %w·w−1). The strong selective pressure of low pH towards the growth of Type II methanotrophic bacteria could explain this finding. The genus Methylocystis increased its abundance from 34 % up to 85 and 90 % at pH 5.5 and 7, respectively. On the contrary, Methylocystis was less abundant in the community enriched at pH 8.5 (14 %). The accumulation of intracellular PHB as energy and carbon storage material allowed the maintenance of high CH4 biodegradation rates during 48 h after complete nitrogen deprivation. The results here obtained demonstrated for the first time a crucial and multifactorial role of pH on the bioconversion performance of CH4 into PHA.

Influence of Nutrient Deprivation on the Antioxidant Capacity and Chemical Profile of Two Diatoms from Genus Chaetoceros.

The limited availability of phosphate, nitrogen and silicon in the growth media affects the growth, cellular processes, and metabolism of diatoms. Silicon deficiency primarily affects diatom morphology, while phosphate deficiency reduces the production of nucleic acids and phospholipids. Differences in pigment and protein composition are mainly due to nitrogen deficiency. In this study, Chaetoceros socialis and Chaetoceros costatus were cultured under phosphate, nitrogen, and silicon deprivation conditions. The diatom biomass was collected during the stationary growth phase and extracted with 70% ethanol under ultrasonication. The chemical profiles of the extracts were analyzed by high-performance liquid chromatography with high-resolution mass spectrometry with electrospray ionisation (UHPLC-ESI-HRMS), while the antioxidant capacity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and oxygen radical absorbance capacity (ORAC) assays. Pigments, fatty acids, sterols, and derivatives were detected in both species. The total phenolic content in the extracts ranged from 46.25 ± 1.08 to 89.38 ± 6.21 mg of gallic acid equivalent (GAE)/L and from 29.58 ± 1.08 to 54.17 ± 1.18 mg GAE/L. for C. costatus and C. socialis, respectively. Antioxidant activity was higher in C. costatus extracts, especially those obtained from nitrogen-deprived media. The results of this study contribute to the existing knowledge and the ongoing efforts to overcome application and commercialization barriers of microalgae for wide-ranging potential in different industries.

Exogenous Trehalose Improves Growth, Glycogen and Poly-3-Hydroxybutyrate (PHB) Contents in Photoautotrophically Grown Arthrospira platensis under Nitrogen Deprivation.

Glycogen and poly-3-hydroxybutyrate (PHB) are excellent biopolymer products from cyanobacteria. In this study, we demonstrate that nitrogen metabolism is positively influenced by the exogenous application of trehalose (Tre) in Arthrospira platensis under nitrogen-deprived (-N) conditions. Cells were cultivated photoautotrophically for 5 days under -N conditions, with or without the addition of exogenous Tre. The results revealed that biomass and chlorophyll-a content of A. platensis experienced enhancement with the addition of 0.003 M and 0.03 M Tre in the -N medium after one day, indicating relief from growth inhibition caused by nitrogen deprivation. The highest glycogen content (54.09 ± 1.6% (w/w) DW) was observed in cells grown for 2 days under the -N + 0.003 M Tre condition (p < 0.05), while the highest PHB content (15.2 ± 0.2% (w/w) DW) was observed in cells grown for 3 days under the -N + 0.03 M Tre condition (p < 0.05). The RT-PCR analysis showed a significant increase in glgA and phaC transcript levels, representing approximately 1.2- and 1.3-fold increases, respectively, in A. platensis grown under -N + 0.003 M Tre and -N + 0.03 M Tre conditions. This was accompanied by the induction of enzyme activities, including glycogen synthase and PHA synthase with maximal values of 89.15 and 0.68 µmol min-1 mg-1 protein, respectively. The chemical structure identification of glycogen and PHB from A. platensis was confirmed by FTIR and NMR analysis. This research represents the first study examining the performance of trehalose in promoting glycogen and PHB production in cyanobacteria under nitrogen-deprived conditions.

Analysis of fate determination of vegetative cells with reduced phycocyanin content in a multicellular cyanobacterium using Raman microscopy.

The one-dimensional multicellular cyanobacterium Anabaena sp. PCC 7120 exhibits two different cell types under nitrogen-deprived conditions. We found that the intensity of the Raman band at 1,629 cm -1 , which is associated with phycocyanin, was higher in undifferentiated cells (vegetative cells) than in differentiated cells (heterocysts). We observed cells whose band intensity at 1,629 cm -1 was statistically lower than that of vegetative cells, and named them "proheterocysts". We found that proheterocysts did not necessarily differentiate, and could divide or revert to being vegetative cells, as defined by having a higher band intensity at 1,629 cm -1 .

Halotolerant bacterial endophyte Bacillus velezensis CBE mediates abiotic stress tolerance with minimal transcriptional modifications in Brachypodium distachyon.

Nitrogen and water are the primary resources limiting agricultural production worldwide. We have demonstrated the ability of a novel halotolerant bacterial endophyte, Bacillus velezensis CBE, to induce osmotic stress tolerance in Brachypodium distachyon under nitrogen-deprived conditions. Additionally, we aimed to identify the molecular factors in plants that contribute to the beneficial effects induced by B. velezensis CBE in B. distachyon. To achieve this, we conducted transcriptomic profiling using RNA-seq on 18-day-old B. distachyon seedlings treated with B. velezensis CBE in the presence or absence of available nitrogen, with and without osmotic stress. These profiles were then compared to those obtained from B. distachyon treated with known plant growth-promoting bacterial strains, Azospirillum brasilense Cd and Azoarcus olearius DQS4, under the same growth conditions. We identified differentially expressed genes (DEGs) in response to the combinations of bacterial strains and stress treatments. Interestingly, only 73 transcripts showed significant differential expression in B. velezensis CBE-treated plants under stress conditions, compared to 1,078 DEGs in plants treated with A. brasilense Cd and 2,015 DEGs in A. olearius DQS4. Our findings suggest that the novel endophyte B. velezensis CBE mediates osmotic stress tolerance in B. distachyon through the fine-tuning of molecular mechanisms with minimal transcriptional modifications.

Novel Insight of Nitrogen Deprivation Affected Lipid Accumulation by Genome-Wide Lactylation in Nannochloropsis oceanica.

Lysine lactylation (kla) as a post-translational modification (PTM) is linked to chromatin remodeling and gene transcription. Nitrogen stress can induce triacylglyceride (TAG) accumulation in most microalgae. To reveal lipid synthesis from another layer, the 4D label-free proteome method was used to track lactylation modifications and protein expression profiles after 3 days of nitrogen deprivation in Nannochloropsis oceanica to analyze the function and prevalence of lactylation. Under nitrogen deprivation (ND) and nitrogen repletion (NR) cultivation conditions, we mapped the proteome and lactylome of proliferating microalgal cells. The results showed that there were 5203 proteins identified and 1245 proteins differentially expressed; among them, 664 were upregulated and 581 were downregulated. The N. oceanica strain also has a total of 868 lactylation sites, which were found on 379 proteins. The lactylated proteins were linked to biological activities as diverse as lipid metabolism, carbon fixation, photosynthesis, glycolysis, and the tricarboxylic acid (TCA) cycle and were found in various subcellular compartments. Under ND, 414 specific lactylation sites were found in 252 proteins. More interestingly, under ND vs NR, lactylation showed a significantly upregulated trend. This study provides a foundation for the future epigenetic enhancement of microalgal cells by providing the first dataset of lactylome in Nannochloropsis.

The genomes of Vischeria oleaginous microalgae shed light on the molecular basis of hyper-accumulation of lipids

BackgroundWith the urgent need to reduce carbon emissions, and the dwindling reserves of easily exploitable fossil fuel, microalgae-based biofuels that can be used for transport systems and CO2 abatement have attracted great attention worldwide in recent years. One useful characteristic of microalgae is their ability to accumulate high levels of lipid content, in particular under conditions of nitrogen deprivation, with numerous species identified so far. However, a trade-off between levels of lipid accumulation and biomass productivity hinders the commercial applicability of lipids from microalgae. Here, we sequenced the genomes of Vischeria sp. CAUP H4302 and Vischeria stellata SAG 33.83, which can accumulate high content of lipids rich in nutraceutical fatty acids and with excellent biomass yield in nitrogen-limiting culture.ResultsA whole-genome duplication (WGD) event was revealed in V. sp. CAUP H4302, which is a rare event in unicellular microalgae. Comparative genomic analyses showed that a battery of genes encoding pivotal enzymes involved in fatty acids and triacylglycerol biosynthesis, storage polysaccharide hydrolysis, and nitrogen and amino acid-related metabolisms are expanded in the genus Vischeria or only in V. sp. CAUP H4302. The most highlighted is the expansion of cyanate lyase genes in the genus Vischeria, which may enhance their detoxification ability against the toxic cyanate by decomposing cyanate to NH3 and CO2, especially under nitrogen-limiting conditions, resulting in better growth performance and sustained accumulation of biomass under the aforementioned stress conditions.ConclusionsThis study presents a WGD event in microalgae, providing new insights into the genetic and regulatory mechanism underpinning hyper-accumulation of lipids and offering potentially valuable targets for future improvements in oleaginous microalgae by metabolic engineering.

Photorespiration plays an important role in H2 production by marine Chlorella pyrenoidosa

Marine Chlorella pyrenoidosa was previously reported to produce H2 under nitrogen-deprived condition. In this study, the role of photorespiration in H2 production by C. pyrenoidosa was investigated. It was found that the expression level of genes encoding three enzymes involved in photorespiration decreased, but the expression level of gene encoding glutamate synthase increased in nitrogen-deprived culture compared with control, indicating a complex adjustment of photorespiration activity during H2 production by C. pyrenoidosa. The use of a photorespiration inhibitor led not only to a decrease in H2 production by 15 % under nitrogen deprivation condition, but also to a 16 % decrease in O2 evolution in the complete medium. Moreover, it was demonstrated that the total photosynthetic oxygen evolution and Fv/Fm declined when photorespiration was blocked. This suggested that, the photorespiration pathway does not directly participate in the production of photosynthetic electrons and the transfer of hydrogen producing electrons. However, it played an important role in photoprotection of PSII during H2 production by C. pyrenoidosa, which enabled PSII to provide more photosynthetic electrons to hydrogenase.

Carotenogenic Activity of Two Hypersaline Greek Dunaliella salina Strains under Nitrogen Deprivation and Salinity Stress

The culture growth and carotenogenic activity of two Greek Dunaliella salina strains (AthU-Al D30 and AthU-Al D31) under stress conditions are investigated herein, with emphasis on β-carotene production as well as on lutein and zeaxanthin. In particular, the strains were cultivated in “standard” conditions (60 ‰ salinity and 1.18 M of NaNO3), under salinity stress conditions (160 ‰ salinity and 1.18 M of NaNO3) and under nitrogen deprivation conditions (60 ‰ salinity and 0 M of NaNO3). In addition to the two Greek strains, the D. salina CCAP 19/18 strain, which has been extensively studied regarding carotenogenesis, is included in this study as a reference. All three strains were found to produce increased amounts of β-carotene when cultivated under nitrogen deprivation conditions, while the AthU-Al D31 strain also produced high amounts of carotenoids under salt stress. The HPLC carotenoid profiles of the strains revealed reduced production of lutein in nitrogen deprivation conditions, coupled with a high production of β-carotene. The strains exhibited various responses in terms of carotenogenic activity, indicating an intraspecific variation in the metabolic reactions related to carotenogenesis.

Position-dependent changes in phycobilisome abundance in multicellular cyanobacterial filaments revealed by Raman spectral analysis.

The one-dimensional filamentous cyanobacterium, Anabaena sp. PCC 7120, shows a simple morphological pattern consisting of two distinct cell types under nitrogen-deprived conditions. We found that microbial pigment composition in differentiated (heterocyst) and undifferentiated cells (vegetative cells) can be distinguished using Raman microscopy. The Raman bands associated with phycocyanin and allophycocyanin were of higher intensity in vegetative cells than those in heterocysts. However, these bands had statistically lower intensity in vegetative cells located further away from heterocysts. That is, the pigment composition in individual cells is affected by locational information in a filament.

Physiological and Genetic Regulation for High Lipid Accumulation by Chlorella sorokiniana Strains from Different Environments of an Arctic Glacier, Desert, and Temperate Lake under Nitrogen Deprivation Conditions.

ABSTRACTMicroalgae can adapt to extreme environments with specialized metabolic mechanisms. Here, we report comparative physiological and genetic regulation analyses of Chlorella sorokiniana from different environmental regions of an arctic glacier, desert, and temperate native lake in response to N deprivation, for screening the optimal strain with high lipid accumulation. Strains from the three regions showed different growth and biochemical compositions under N deprivation. The arctic glacier and desert strains produced higher soluble sugar content than strains from the native lake. The arctic glacier strains produced the highest levels of lipid content and neutral lipids under N deprivation compared with strains from desert and native lake. At a molecular level, the arctic strain produced more differentially expressed genes related to fatty acid biosynthesis, glycolysis gluconeogenesis, and glycerolipid metabolism. The important functional genes acetyl coenzyme A (acetyl-CoA) carboxylase (ACCase), fatty acid synthase complex, pyruvate dehydrogenase component, and fatty acyl-acyl carrier protein (acyl-ACP) thioesterase were highly expressed in arctic strains. More acetyl-CoA was produced from glycolysis gluconeogenesis and glycerolipid metabolism, which then produced more fatty acid with the catalytic function of ACCase and acyl-ACP thioesterase in fatty acid biosynthesis. Our results indicated that the C. sorokiniana strains from the arctic region had the fullest potential for biodiesel production due to special genetic regulation related to fatty acid synthesis, glycolysis gluconeogenesis, and glycerolipid metabolism.IMPORTANCE It is important to reveal the physiological and genetic regulation mechanisms of microalgae for screening potential strains with high lipid production. Our results showed that Chlorella sorokiniana strains from arctic glacier, desert, and temperate native lake had different growth, biochemical composition, and genetic expression under N deprivation. The strains from an arctic glacier produced the highest lipid content (including neutral lipid), which was related to the genetic regulation of fatty acid biosynthesis, glycolysis gluconeogenesis, and glycerolipid metabolism. The functional genes for acetyl-CoA carboxylase, fatty acid synthase complex, pyruvate dehydrogenase component, and fatty acyl-ACP thioesterase in the three pathways were highly expressed in arctic strains. The revelation of physiological and genetic regulation of strains from different environmental regions will contribute to the microalgae selection for high lipid accumulation.

Lipid turnover and SQUAMOSA promoter-binding proteins mediate variation in fatty acid desaturation under early nitrogen deprivation revealed by lipidomic and transcriptomic analyses in Chlorella pyrenoidosa.

Nitrogen deprivation induces variations in fatty acid desaturation in microalgae, which determines the performance of biodiesel and the nutritional value of bioproducts. However, the detailed scenario and the underlying regulatory mechanism remain unclear. In this study, we attempt to outline these scenario and mechanisms by performing biochemical, lipidomic, and transcriptomic analyses in Chlorella pyrenoidosa and functional characterization of transcription factors in Yarrowia lipolytica. We found that early nitrogen deprivation dramatically reduced fatty acid desaturation without increasing lipid content. The contents of palmitic acid (16:0) and oleic acid (18:1) dramatically increased to 2.14 and 2.87 times that of nitrogen repletion on the second day, respectively. Lipidomic analysis showed the transfer of polyunsaturated fatty acids from phospholipids and glycolipids to triacylglycerols, and an increase in lipid species with 16:0 or 18:1 under nitrogen deprivation conditions. Upregulated stearoyl-ACP desaturase and oleyl-ACP thioesterase promoted the synthesis of 18:1, but restricted acetyl-CoA supply revealed that it was the intensive lipid turnover instead of an attenuated Kennedy pathway that played an important role in the variation in fatty acid composition under early nitrogen deprivation. Finally, two differentially expressed SQUAMOSA promoter-binding proteins (SBPs) were heterologously expressed in Y. lipolytica, demonstrating their role in promoting the accumulation of total fatty acid and the reduction in fatty acid desaturation. These results revealed the crucial role of lipid turnover and SBPs in determining fatty acid desaturation under early nitrogen deprivation, opening new avenues for the metabolic engineering of fatty acid desaturation in microalgae.

Possible involvement of extracellular polymeric substrates of Antarctic cyanobacterium Nostoc sp. strain SO-36 in adaptation to harsh environments.

Cyanobacteria are some of the primary producers in extremely cold biospheres such as the Arctic, Antarctic, and vast ice sheets. Many genera of cyanobacteria are identified from these harsh environments, but their specific mechanisms for cold adaptation are not fully understood. Nostoc sp. strain SO-36 is a cyanobacterium isolated in Antarctica more than 30years ago and regarded as a psychrotolelant species. To determine whether the strain is psychrotolelant or psychrophilic, it was first grown at 30°C and 10°C. The cells grew exponentially at 30°C, but their growth stopped at 10°C, indicating that the strain is only psychrotolerant. Microscopic analysis revealed that the morphology of the cells grown at 30°C was filamentous and differentiated heterocysts, which are specialized cells for gaseous nitrogen fixation under nitrogen-deprived conditions, indicating that the strain can grow diazotrophically. The cells grown at 10°C have a smaller size, shortened filament length and decreased chlorophyll content per cell. At 10°C, the cells are aggregated with extracellular polymeric substrates (EPSs), which is a common mechanism to protect cells from ultraviolet light. These results imply that segmentation into short filaments was induced by photodamage at low temperatures. To fully understand the adaptation mechanisms of Nostoc sp. strain SO-36 for low-temperature conditions, next-generation sequencing analyses were conducted. Complete genome sequence of the strain revealed that it has one main chromosome of approximately 6.8 Mbp with 4 plasmids, including 6855 coding sequences, 48 tRNA genes, 4 copies of rRNA operons, and 5 CRISPR regions. Putative genes for EPS biosynthesis were found to be conserved in Nostocaceae regardless of their habitat. These results provide basic information to understand the adaptation mechanisms at low temperatures, and the strain can be a model organism to analyze adaptation to extreme environments.

Growth characteristics, biohydrogen production and photochemical activity of photosystems in green microalgae Parachlorella kessleri exposed to nitrogen deprivation

The biohydrogen (H2) generation in green microalgae is associated with electron transfer during photosynthesis. In this study, the ability of Parachlorella kessleri RA-002 isolated in Armenia to generate biohydrogen (H2) in TAP and Tamiya media with and without nitrogen deprivation have been determined. Nitrogen deprivation led to a suppression of the growth rate and a decrease in the content of photosynthetic pigments in algae, as well as to increase of H2 yield. The highest H2 yield was found during the growth of algae in TAP and Tamiya media under nitrogen-deprived conditions, which was 4–5 times more compared to control cells grown without nitrogen deprivation. Diuron inhibited this process, indicating that H2 generation was due to PS II activities. The analysis of the effect of nitrogen-deprived conditions on the activity of photosystem I and II has been carried out. Nitrogen deprivation suppressed of photosynthetic activity of PS II (up to 20% compared to control) that facilitated the formation of anaerobic conditions increasing the total yield of H2.

Vegetative cells may perform nitrogen fixation function under nitrogen deprivation in Anabaena sp. strain PCC 7120 based on genome-wide differential expression analysis.

Nitrogen assimilation is strictly regulated in cyanobacteria. In an inorganic nitrogen-deficient environment, some vegetative cells of the cyanobacterium Anabaena differentiate into heterocysts. We assessed the photosynthesis and nitrogen-fixing capacities of heterocysts and vegetative cells, respectively, at the transcriptome level. RNA extracted from nitrogen-replete vegetative cells (NVs), nitrogen-deprived vegetative cells (NDVs), and nitrogen-deprived heterocysts (NDHs) in Anabaena sp. strain PCC 7120 was evaluated by transcriptome sequencing. Paired comparisons of NVs vs. NDHs, NVs vs. NDVs, and NDVs vs. NDHs revealed 2,044 differentially expressed genes (DEGs). Kyoto Encyclopedia of Genes and Genomes enrichment analysis of the DEGs showed that carbon fixation in photosynthetic organisms and several nitrogen metabolism-related pathways were significantly enriched. Synthesis of Gvp (Gas vesicle synthesis protein gene) in NVs was blocked by nitrogen deprivation, which may cause Anabaena cells to sink and promote nitrogen fixation under anaerobic conditions; in contrast, heterocysts may perform photosynthesis under nitrogen deprivation conditions, whereas the nitrogen fixation capability of vegetative cells was promoted by nitrogen deprivation. Immunofluorescence analysis of nitrogenase iron protein suggested that the nitrogen fixation capability of vegetative cells was promoted by nitrogen deprivation. Our findings provide insight into the molecular mechanisms underlying nitrogen fixation and photosynthesis in vegetative cells and heterocysts at the transcriptome level. This study provides a foundation for further functional verification of heterocyst growth, differentiation, and water bloom control.

Enhanced polyhydroxybutyrate (PHB) accumulation in heterotrophically grown Arthrospira platensis under nitrogen deprivation

The non-heterocystous cyanobacterium Arthrospira platensis was investigated for the conditions under which high accumulation of polyhydroxybutyrate (PHB) occurred. Cells grown photoautotrophically under nitrogen-deprived conditions had high accumulation of PHB whose granules were clearly visualized using Nile-Red staining. A further increase of PHB was observed in cells grown photoheterotrophically for 3 days under nitrogen-deprived condition with 0.50%(w/v) acetate supplementation (−N +Act) showing the maximum PHB contents of 19.2 ± 0.5%(w/w DW). This was significantly higher than that in cells grown under the same condition but with either 0.1%(w/v) butyrate, or 0.75%(w/v) glucose, or 0.1%(w/v) propionate supplementation (P < 0.05). RT-PCR analysis revealed that photoheterotrophically grown cells under −N+ Act condition had a significant increase of phaB and phaC transcript levels (P < 0.05) compared with those supplemented with other organic carbons. The representative FTIR spectra of the extracted polymer from A. platensis showed prominent peaks at 1721 cm−1 and 1279 cm−1 for the carbonyl (C=O) stretching of the ester group and asymmetric C–O–C stretching vibration, respectively. This spectral pattern is comparable with that of the standard PHB spectra, thus confirming the chemical nature of the extract. The overall results demonstrated that the enhanced PHB accumulation in A. platensis was attributed to the increased expression of phaB and phaC responsible for increased PHB biosynthesis when cells were grown photoheterotrophically in nitrogen-deprived medium containing organic carbon compounds.

Assessment of Protein Content and Phosphorylation Level in Synechocystis sp. PCC 6803 under Various Growth Conditions Using Quantitative Phosphoproteomic Analysis.

The photosynthetic apparatus and metabolic enzymes of cyanobacteria are subject to various controls, such as transcriptional regulation and post-translational modifications, to ensure that the entire cellular system functions optimally. In particular, phosphorylation plays key roles in many cellular controls such as enzyme activity, signal transduction, and photosynthetic apparatus restructuring. Therefore, elucidating the governing functions of phosphorylation is crucial to understanding the regulatory mechanisms underlying metabolism and photosynthesis. In this study, we determined protein content and phosphorylation levels to reveal the regulation of intracellular metabolism and photosynthesis in Synechocystis sp. PCC 6803; for this, we obtained quantitative data of proteins and their phosphorylated forms involved in photosynthesis and metabolism under various growth conditions (photoautotrophic, mixotrophic, heterotrophic, dark, and nitrogen-deprived conditions) using targeted proteomic and phosphoproteomic analyses with nano-liquid chromatography-triple quadrupole mass spectrometry. The results indicated that in addition to the regulation of protein expression, the regulation of phosphorylation levels of cyanobacterial photosynthetic apparatus and metabolic enzymes was pivotal for adapting to changing environmental conditions. Furthermore, reduced protein levels of CpcC and altered phosphorylation levels of CpcB, ApcA, OCP, and PsbV contributed to the cellular response of the photosynthesis apparatus to nitrogen deficiency.