Nitrogen Deprivation Research Articles

Enhancement of ketocarotenoid production using the heterologous orange protein from Ipomoea batatas in indigenous microalga Ettlia sp.

Astaxanthin, one of the most powerful natural antioxidants, is used in high-value industries such as those of cosmetics, nutraceuticals, and food products derived from microalgae. In this study, Ettlia sp. mutants were generated by expressing two types of heterologous orange proteins, IbOr and IbOr-R96H, in which the 96th arginine of IbOr was substituted with histidine derived from Ipomoea batatas to enhance astaxanthin production. The Ett-IbOr-R96H mutant showed a 2.4-fold increase in β-carotene content compared to the wildtype (4.59 mg g−1DCW), reaching 10.82 mg g−1 under high-light conditions via two-phase cultivation. Under the stress treatment combination of high-light intensity and nitrogen deprivation, total carotenoid content increased to 17.24 mg L−1 and 21.94 mg L−1 in Ett-IbOr and Ett-IbOr-R96H, respectively. The astaxanthin and canthaxanthin contents in Ett-IbOr-R96H was 4.89 mg L−1 and 0.47 mg L−1, respectively, which were 1.8- and 1.5-fold higher, respectively, than those in Ett-IbOr. Additionally, photosynthetic efficiency (Fv/Fm) recovered in Ett-IbOr-R96H under dual-stress conditions compared to the wildtype while reactive oxygen species levels decreased throughout the cultivation period. Our findings suggest that the heterologous IbOr expression in Ettlia sp. may be an effective approach for enhancing the production of ketocarotenoids and improving stress resistance for industrial applications.

Enhancing methane yield from microalgae: abiotic stress and cells disruption with quartz powder

This study investigates the biochemical methane potential (BMP) of microalgal biomass, introducing a novel cells disruption method using quartz powder (SiO2). A two-phase algae cultivation, involving nitrogen deprivation and salinity shifts, was employed to biochemically modify the biomass of two brackish green algae strains, Chlorella vulgaris and Monoraphidium contortum, enhancing their methane (CH4) production potential. Mechanical disruption of the algal cells further increased BMP, with C. vulgaris yielding 305 mL CH4/g volatile solids (VS) and M. contortum reaching 324 mL CH4/g VS, reflecting respective increases of 51 % and 86 %. The integration of this efficient mechanical cell disruption method with a simple, stress-based cultivation strategy presents significant potential for enhancing the methane yield of microalgal biomass.

Metabolic profiling reveal changes in shoots and roots of nitrogen-deficient tea plants (Camellia sinensis cv. Jinxuan)

Tea plants require high levels of nitrogen for optimal growth. However, delayed or insufficient nitrogen supply often induces nitrogen deficiency stress. This stress can adversely affect the plant's growth, development, and metabolic processes. Therefore, studying how tea plants respond to nitrogen deficiency conditions is crucial for enhancing our understanding and management of their growth environment. In this study, we utilized High-Performance Liquid Chromatography (HPLC), Gas Chromatography-Mass Spectrometry (GC–MS), and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS) techniques to analyze the non-volatile and volatile components of both shoots and roots of the 'Jing Xuan (JX)' tea variety under nitrogen deprivation for 30 days (Nd). The results revealed that nitrogen deprivation led to a deceleration in shoot growth of 'JX', accompanied by reduced chlorophyll and water-soluble extract contents, with nitrogen accumulating predominantly in the roots. Significant decreases were observed in caffeine content in shoots and in the total amounts of theanine and catechins in both shoots and roots, while epigallocatechin gallate (EGCG) accumulation increased in shoots. Moreover, we identified 65 volatile components in shoots, including a notable decrease in floral and fruity compounds such as trans-β-Ionone, Geranylacetone, and α-Ionone. The biosynthesis of amino acids and phenylpropane metabolism are more active pathways under nitrogen stress. Nitrogen deficiency led to reduced levels of amino acids, carboxylic acids, flavonoids, steroids, and their derivatives in roots, with the most notable decline observed in carboxylic acids, typified by Leucine, l-Aspartate, and l-histidine. In contrast, shoots accumulate carbohydrates and phenolic compounds, mainly sucrose. Additionally, metabolites such as Myristic acid and 3-Dehydroshikimate could be a critical indicator of stress levels. These findings enhance our understanding of tea plant growth and metabolism under nitrogen deficiency, providing valuable insights for breeding nitrogen-efficient tea varieties.

Disrupted H2 synthesis combined with methyl viologen treatment inhibits photosynthetic electron flow to synergistically enhance glycogen accumulation in the cyanobacterium Synechocystis sp. PCC 6803.

Under nitrogen deprivation (-N), cyanobacterium Synechocystis sp. PCC 6803 exhibits growth arrest, reduced protein content, and remarkably increased glycogen accumulation. However, producing glycogen under this condition requires a two-step process with cell transfer from normal to -N medium. Metabolic engineering and chemical treatment for rapid glycogen accumulation can bypass the need for two-step cultivation. For example, recent studies indicate that individually disrupting hydrogen (H2) or poly(3-hydroxybutyrate) (PHB) synthesis, or treatment with methyl viologen (MV), effectively increases glycogen accumulation in Synechocystis. Here we explore the effects of disrupted H2 or poly(3-hydroxybutyrate) synthesis, together with MV treatment to on enhanced glycogen accumulation in Synechocystis grown in normal medium. Wild-type cells without MV treatment exhibited low glycogen content of less than 6% w/w dry weight (DW). Compared with wild type, disrupting PHB synthesis combined with MV treatment did not increase glycogen content. Disrupted H₂ production without MV treatment yielded up to 11% w/w DW glycogen content. Interestingly, when combined, disrupted H2 production with MV treatment synergistically enhanced glycogen accumulation to 51% and 59% w/w DW within 3 and 7days, respectively. Metabolomic analysis suggests that MV treatment mediated the conversion of proteins into glycogen. Metabolomic and transcriptional-expression analysis suggests that disrupted H2 synthesis under MV treatment positively influenced glycogen synthesis. Disrupted H₂ synthesis under MV treatment significantly increased NADPH levels. This increased NADPH content potentially contributed to the observed enhancements in antioxidant activity against MV-induced oxidants, O2 evolution, and metabolite substrates levels for glycogen synthesis in normal medium, ultimately leading to enhanced glycogen accumulation in Synechocystis. KEY MESSAGE: Combining disrupted hydrogen-gas synthesis and the treatment by photosynthesis electron-transport inhibitor significantly enhance glycogen production in cyanobacteria.

Autotrophic poly-3-hydroxybutyrate accumulation in Cupriavidus necator for sustainable bioplastic production triggered by nutrient starvation

Cupriavidus necator is a facultative chemolithoautotrophic bacterium able to convert carbon dioxide into poly-3-hydroxybutyrate. This is highly promising as the conversion process allows the production of sustainable and biodegradable plastics. Poly-3-hydroxybutyrate accumulation is known to be induced by nutrient starvation, but information regarding the optimal stress conditions controlling the process is still heterogeneous and fragmentary. This study presents a comprehensive comparison of the effects of nutrient stress conditions, namely nitrogen, hydrogen, phosphorus, oxygen, and magnesium deprivation, on poly-3-hydroxybutyrate accumulation in C. necator DSM545. Nitrogen starvation exhibited the highest poly-3-hydroxybutyrate accumulation, achieving 54% of total cell dry weight after four days of nutrient stress, and a carbon conversion efficiency of 85%. The gas consumption patterns indicated flexible physiological mechanisms underlying polymer accumulation and depolymerization. These findings provide insights into strategies for efficient carbon conversion into bioplastics, and highlight the key role of C. necator for future industrial-scale applications.

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.

Red light induces starch accumulation in Chlorella vulgaris without affecting photosynthesis efficiency, unlike abiotic stress

Microalgae show great promise as sources of starch, one of the most widely consumed macromolecules. In this study, we evaluated the impact of three starch-inducing factors, namely nitrogen deprivation, supra-optimal temperature, and red light, on the physiology and starch accumulation capacity of Chlorella vulgaris. This starch accumulation was monitored by measuring the total carbohydrate content and transmission electron microscopy (TEM) imaging. Nitrogen deprivation and a supra-optimal temperature of 39 °C resulted in carbohydrate contents of 69.7 and 64.3 % of dry weight (DW) respectively. This constituted a 5.3- and 3.3-fold increase in carbohydrate productivity compared to the control, after 4 days of cultivation. During this period, carbohydrates represented over 80 % of the produced material (DW basis). However, nitrogen deprivation and supra-optimal temperature were accompanied by extensive stress, leading to lower cell division rates and damage to the photosynthetic apparatus. Red light illumination resulted in a more moderate production of carbohydrates. After 4 days of cultivation, the carbohydrate content reached 46.8 %, representing a 3.0-fold increase in productivity compared to control. The composition of the starch formed under red light was surprisingly poor in amylose, similar to transitory-type starch rather than storage starch. Most notably, the starch accumulation under red light was sustained over 7 days without affecting the rate of cell division and quantum yield efficiency. To the best of our knowledge, red light is the only factor reported so far to induce a significant starch accumulation without hindering cell division and photosynthesis efficiency, even after long-term exposure (7 days). Furthermore, all three conditions induced a cell wall thickening, albeit without affecting the recovery of accumulated starch by high-pressure homogenization. These results highlight the potential of red light as a starch inducer in Chlorella vulgaris and open up perspectives for the production of starch-based bioplastics from microalgae.

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.

Changes in the Root Architecture of Oil Palm Seedlings in Response to Nitrogen Starvation

Oil palm (Elaeis guineensis) is a widely cultivated crop known for its high oil yield. It is cultivated extensively across tropical regions, notably in Southeast Asia, Africa, and Latin America. It plays a vital role in global vegetable oil supply, meeting approximately 35% of the world’s demand. However, the expansion of oil palm plantations often involves the utilization of degraded soils where nutrient availability, particularly nitrogen, is limited, posing challenges to plant growth and productivity. Roots are crucial in responding to nitrogen deficiency by adjusting their growth and distribution; however, research on root system distribution patterns in oil palm still needs to be completed. This study analyzes the root system architecture using RhizoVision Explorer, a 2D root image processing software while assessing its relationship with nitrogen availability across two commercial cultivars: Deli × La Mé of African oil palm (Elaeis guineensis) and the interspecific hybrid O×G Coari × La Mé. Our findings reveal significant associations between eight root traits in oil palm seedlings and treatments with and without nitrogen availability. Notably, total root morphology (length, surface area, and volume), rotation angle, solidity, and hole characteristics decreased under nitrogen deprivation, whereas surface angle frequency increased. We highlight the variability of these traits across cultivars, suggesting genetic dependence and potential utility in breeding programs. Moreover, interactions observed in primary root morphology and hole size indicate greater differences between control and nitrogen-treated groups in C × LM than in D × LM cultivars. On the other hand, cultivar differences, regardless of nitrogen availability, influenced lateral root morphology, while nitrogen availability, irrespective of cultivar, affected inclined angle frequency. Significant differences were observed in growth and development parameters such as root and shoot biomass, root-to-shoot ratio, and leaf emission numbers between nitrogen-optimal and nitrogen-starved conditions. Nitrogen significantly affects root architecture and plant growth in oil palm, particularly in the C × LM cultivar during the nursery stage.

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.

Comparison of sulfur and nitrogen deprivation effects on photosynthetic pigments, polyphenols, photosystems activity and H2 generation in Chlorella vulgaris and Parachlorella kessleri

Biohydrogen (H2) production by green algae has great biotechnological potential. This study aimed to analyze photosynthetic pigment levels, photosynthetic systems activity, total phenolic/flavonoid content, and H2 generation in Chlorella vulgaris IBCE C-19 and Parachlorella kessleri MDC6524 in response to sulfur (S) and nitrogen (N) deprivation. The results showed that S and N deficiency caused a growth rate decrease in both cultures that was accompanied by a decrease in the photosynthetic pigments levels, PS II activity, and polyphenols synthesis. In addition, S and N deprivation led to an increase in the H2 generation. Moreover, the H2 photoproduction by P. kessleri was higher compared to that of C. vulgaris under N- and S-deprived anaerobic conditions. DCMU inhibited the H2 production by both algae showing the involvement of PS II in this process. The findings contribute to the ongoing research on the development of strategies to optimize algae cultivation for enhanced H2 yield.

Optimizing light regimes for neutral lipid accumulation in Dunaliella salina MCC 43: a study on physiological status and carbon allocation.

Dunaliella salina is a favourable source of high lipid feedstock for biofuel and medicinal chemicals. Low biomass output from microalgae is a significant barrier to industrial-scale commercialisation. The current study aimed to determine how photosynthetic efficiency, carbon fixation, macromolecular synthesis, accumulation of neutral lipids, and antioxidative defence (ROS scavenging enzyme activities) of D. salina cells were affected by different light intensities (LI) (50, 100, 200, and 400µmolm-2s-1). The cells when exposed to strong light (400µmolm-2s-1) led to reduction in chlorophyll a but the carotenoid content increased by 19% in comparison to the control (LI 100). The amount of carbohydrate changed significantly under high light and in spite of stress inflicted on the cells by high irradiation, a considerable increase in activity of carbonic anhydrase and fixation rate of CO2 were recorded, thus, preserving the biomass content. The high light exposed biomass when subjected to nitrogen-deficient medium led to increase in lipid content (59.92% of the dry cell weight). However, neutral lipid made up 78.26% of the total lipid while other lipids like phospholipid and glycolipid content decreased, showing that the lipid was redistributed in these cells under nitrogen deprivation, making the organism more appropriate for biodiesel/jet fuel use. Although D. salina cells had a relatively longer generation time (3.5 d) than other microalgal cells, an economic analysis concluded that the amount of carotenoid they produced and the quality of their lipids made them more suited for commercialization.

Low nitrogen status affects isoflavonoid production and flavonol decoration in Lotus corniculatus

Nitrogen availability is key factor for plant metabolism and has a strong impact on the biosynthesis of secondary plant metabolites. A number of these metabolites are involved in the response to stress. In this paper, we investigated the effect of different N conditions in leaves of Lotus corniculatus. Low nitrogen conditions strongly enhanced the biosynthesis of phenolic compounds in leaves, inducing the expression genes encoding for key enzymes of phenolic metabolism such as phenylalanine ammonia lyase, chalcone synthase and isoflavone synthase. Kaempferol and quercetin glycosides as well as isoflavanes accumulated in leaves. Surprisingly, if the nitrogen availability was limited, but without nitrogen deprivation symptoms, an increase in the levels of phenylpropane acids and a decrease in the levels of kaempferol glycosides was detected. Especially the decrease in malonylated kaempferol derivatives and the kaempferol glycosides substituted by rhamnose in the 3-O position. The L. corniculatus malonyltransferase 1 gene is likely involved in the malonylation of the kaempferol glycosides.

Iron excess and nitrogen deprivation influence photosynthetic metabolism in grasses used for mineland rehabilitation

Iron excess and nitrogen deprivation influence photosynthetic metabolism in grasses used for mineland rehabilitation

Spatio-temporal coherence of circadian clocks and temporal control of differentiation in Anabaena filaments.

Circadian clock arrays in multicellular filaments of the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 display remarkable spatio-temporal coherence under nitrogen-replete conditions. To shed light on the interplay between circadian clocks and the formation of developmental patterns, we followed the expression of a clock-controlled gene under nitrogen deprivation, at the level of individual cells. Our experiments showed that differentiation into heterocysts took place preferentially within a limited interval of the circadian clock cycle, that gene expression in different vegetative intervals along a developed filament was discoordinated, and that the circadian clock was active in individual heterocysts. Furthermore, Anabaena mutants lacking the kaiABC genes encoding the circadian clock core components produced heterocysts but failed in diazotrophy. Therefore, genes related to some aspect of nitrogen fixation, rather than early or mid-heterocyst differentiation genes, are likely affected by the absence of the clock. A bioinformatics analysis supports the notion that RpaA may play a role as master regulator of clock outputs in Anabaena, the temporal control of differentiation by the circadian clock and the involvement of the clock in proper diazotrophic growth. Together, these results suggest that under nitrogen-deficient conditions, the clock coherent unit in Anabaena is reduced from a full filament under nitrogen-rich conditions to the vegetative cell interval between heterocysts.IMPORTANCECircadian clocks, from unicellular organisms to animals, temporally align biological processes to day and night cycles. We study the dynamics of a circadian clock-controlled gene at the individual cell level in the multicellular filamentous cyanobacterium Anabaena, under nitrogen-stress conditions. Under these conditions, some cells along filaments differentiate to carry out atmospheric nitrogen fixation and lose their capability for oxygenic photosynthesis. We found that clock synchronization is limited to organismic units of contiguous photosynthetic cells, contrary to nitrogen-replete conditions in which clocks are synchronized over a whole filament. We provided evidence that the circadian clock regulates the process of differentiation, allowing it to occur preferentially within a limited time window during the circadian clock period. Lastly, we present evidence that the signal from the core clock to clock-regulated genes is conveyed in Anabaena as in unicellular cyanobacteria.

Lipid accumulation mechanism of Amphora coffeaeformis under nitrogen deprivation and its application as a feed additive in Carassius auratus aquaculture

BackgroundAmphora coffeaeformis, a unicellular diatom, can significantly accumulate lipids under nitrogen (N) limitation. However, the molecular mechanism underlying lipid accumulation in A. coffeaeformis remains unknown and its application development is lagging.ResultsThis work analyzed the lipid composition of A. coffeaeformis under N deprivation and investigated its mechanism underlying lipid accumulation using RNA-seq. The results showed that the total lipid content of A. coffeaeformis increased from 28.22 to 44.05% after 5 days of N deprivation, while the neutral lipid triacylglycerol (TAG) content increased from 10.41 to 25.21%. The transcriptional profile showed that N deprivation induced wide-ranging reprogramming of regulation and that most physiological activities were repressed, while the upregulation of glycerol-3-phosphate acyltransferase directly determined TAG accumulation. Moreover, we explored the effect of A. coffeaeformis as a food additive on the lipid composition of crucian carp. The results showed that the contents of unsaturated fatty acids in the meat of fish supplemented with A. coffeaeformis were significantly increased, indicating its potential application in animal nutrition for improving meat quality indicators.ConclusionThe findings shed light on the molecular mechanisms of neutral lipid accumulation and revealed the key genes involved in lipid metabolism in A. coffeaeformis. Moreover, we also confirmed that A. coffeaeformis can be used as feed additive for improving the lipid composition of crucian carp meat, which provided evidence for the biotechnology application of this high-oil microalgae.

The combination of uridine and nitrogen-deprivation promotes the efficient formation of astaxanthin-rich motile cells in Haematococcus pluvialis

Astaxanthin accumulation in Haematococcus pluvialis typically occurs alongside the formation of secondary cell wall (SCW), hindering astaxanthin extraction and bio-accessibility. A potential solution lies in cultivating astaxanthin-rich motile cells lacking SCW. This study explored the influence and underlying mechanism of nitrogen-deprivation (ND) on SCW formation and established a connection between pyrimidine metabolism and SCW development. Then, various pyrimidine and ND combinations were examined to cultivate astaxanthin-rich motile cells. The results indicated that, compared to the nitrogen-replete group, the combination of uridine and ND increased the proportion of motile cells by 25–33 times, achieving 95 %, and enhanced astaxanthin yield by 26.52 %. Moreover, the efficiency of astaxanthin extraction from intact, wet motile cells was 91 % – 95 %, which was 5.6–9.0 times that from non-motile cells. This study not only presents a promising method for producing astaxanthin-rich motile cells in H. pluvialis but also provides insights into the relationship between pyrimidine metabolism and SCW development.

Optimal Laboratory Cultivation Conditions of Limnospira maxima for Large-Scale Production.

Cultivating Limnospira maxima, renowned for its abundant proteins and valuable pigments, faces substantial challenges rooted in the limited understanding of its optimal growth parameters, associated high costs, and constraints in the procurement of traditional nitrogen sources, particularly NaNO3. To overcome these challenges, we conducted a comprehensive 4 × 3 factorial design study. Factors considered included white, red, blue, and yellow light spectra, along with nitrogen sources NaNO3 and KNO3, as well as a nitrogen-free control, for large-scale implementation. Optimal growth, measured by Optical Density, occurred with white and yellow light combined with KNO3 as the nitrogen source. These conditions also increased dry weight and Chl-a content. Cultures with nitrogen deprivation exhibited high values for these variables, attributed to carbon accumulation in response to nitrogen scarcity. Phycocyanin, a crucial pigment for nutrition and industry, reached its highest levels in cultures exposed to white light and supplemented with KNO3, with an impressive content of 384.11 g kg-1 of dry weight. These results highlight the efficacy and cost-efficiency of using a combination of white light and KNO3 for large-scale L. maxima cultivation. This strategy offers promising opportunities to address global food security challenges and enhance the production of industrially relevant pigments.