Nitrogen Starvation Conditions Research Articles

Optimal nitrogen source for the recovery of phycoerythrin in discolored Rhodomonas sp.

Rhodomonas species are of interest as excellent aquaculture feed because of their ideal biochemical composition, especially their high protein content, resulting from the presence of intracellular phycoerythrin (PE). One of the most crucial issue with this species as a feed source is PE degradation under ambient nitrogen starvation, which leads to unpredictable growth defects and reduced nutritional value associated with cell discoloration. The aim of this study was to propose an optimal nitrogen source for the recovery of intracellular PE in discolored Rhodomonas sp. We supplied three different nitrogen sources, i.e., nitrate, ammonium, and urea, to PE-degraded cells and monitored changes in PE content and the expression of related genes. Supplementation with each nitrogen source increased cell density, with nitrate exhibiting the highest specific growth rate of 0.30 ± 0.02 day−1. The maximum PE content in each nitrogen group at 48 h after N supplementation was about 3.8 pg cell−1 with no major difference between groups. In contrast, the highest relative PE recovery rate of 5.73 ± 0.18-fold and the highest PE productivities of 0.32 ± 0.01 μg mL−1 h−1 were obtained in the nitrate group. Nitrogen transporter genes, including RhNrt2, RhAmt1, and RhDur3, were upregulated under nitrogen-starvation conditions, demonstrating the ability of this species to utilize each of the tested nitrogen sources. Furthermore, RhCpeb, which serves as a molecular indicator of PE biosynthesis, was significantly upregulated in all nitrogen-supplemented groups, with the strongest and fastest increase in expression in the nitrate group, emphasizing the potential of nitrate as the most favorable nitrogen source for PE recovery in this species. Our findings pave the way for informed strategies for the stable management and production of Rhodomonas species as high-protein feed in aquaculture.

Polyphosphate affects cytoplasmic and chromosomal dynamics in nitrogen-starved Pseudomonas aeruginosa

Polyphosphate (polyP) synthesis is a ubiquitous stress and starvation response in bacteria. In diverse species, mutants unable to make polyP have a wide variety of physiological defects, but the mechanisms by which this simple polyanion exerts its effects remain unclear. One possibility is that polyP's many functions stem from global effects on the biophysical properties of the cell. We characterize the effect of polyphosphate on cytoplasmic mobility under nitrogen-starvation conditions in the opportunistic pathogen Pseudomonas aeruginosa. Using fluorescence microscopy and particle tracking, we quantify the motion of chromosomal loci and cytoplasmic tracer particles. In the absence of polyP and upon starvation, we observe a 2- to 10-fold increase in mean cytoplasmic diffusivity. Tracer particles reveal that polyP also modulates the partitioning between a "more mobile" and a "less mobile" population: Small particles in cells unable to make polyP are more likely to be "mobile" and explore more of the cytoplasm, particularly during starvation. Concomitant with this larger freedom of motion in polyP-deficient cells, we observe decompaction of the nucleoid and an increase in the steady-state concentration of ATP. The dramatic polyP-dependent effects we observe on cytoplasmic transport properties occur under nitrogen starvation, but not carbon starvation, suggesting that polyP may have distinct functions under different types of starvation.

Glycogen Granules Are Degraded by Non-Selective Autophagy in Nitrogen-Starved Komagataella phaffii.

Autophagy was initially recognized as a bulk degradation process that randomly sequesters and degrades cytoplasmic material in lysosomes (vacuoles in yeast). In recent years, various types of selective autophagy have been discovered. Glycophagy, the selective autophagy of glycogen granules, is one of them. While autophagy of glycogen is an important contributor to Pompe disease, which is characterized by the lysosomal accumulation of glycogen, its selectivity is still a matter of debate. Here, we developed the Komagataella phaffii yeast as a simple model of glycogen autophagy under nitrogen starvation conditions to address the question of its selectivity. For this, we turned the self-glucosylating initiator of glycogen synthesis, Glg1, which is covalently bound to glycogen, into the Glg1-GFP autophagic reporter. Our results revealed that vacuolar delivery of Glg1-GFP and its processing to free GFP were strictly dependent on autophagic machinery and vacuolar proteolysis. Notably, this process was independent of Atg11, the scaffold protein common for many selective autophagy pathways. Importantly, the non-mutated Glg1-GFP (which synthesizes and marks glycogen) and mutated Glg1Y212F-GFP (which does not synthesize glycogen and is degraded by non-selective autophagy as cytosolic Pgk1-GFP) were equally well delivered to the vacuole and had similar levels of released GFP. Therefore, we concluded that glycogen autophagy is a non-selective process in K. phaffii yeast under nitrogen starvation conditions.

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.

Phosphate starvation decouples cell differentiation from DNA replication control in the dimorphic bacterium Caulobacter crescentus.

Upon nutrient depletion, bacteria stop proliferating and undergo physiological and morphological changes to ensure their survival. Yet, how these processes are coordinated in response to distinct starvation conditions is poorly understood. Here we compare the cellular responses of Caulobacter crescentus to carbon (C), nitrogen (N) and phosphorus (P) starvation conditions. We find that DNA replication initiation and abundance of the replication initiator DnaA are, under all three starvation conditions, regulated by a common mechanism involving the inhibition of DnaA translation. By contrast, cell differentiation from a motile swarmer cell to a sessile stalked cell is regulated differently under the three starvation conditions. During C and N starvation, production of the signaling molecules (p)ppGpp is required to arrest cell development in the motile swarmer stage. By contrast, our data suggest that low (p)ppGpp levels under P starvation allow P-starved swarmer cells to differentiate into sessile stalked cells. Further, we show that limited DnaA availability, and consequently absence of DNA replication initiation, is the main reason that prevents P-starved stalked cells from completing the cell cycle. Together, our findings demonstrate that C. crescentus decouples cell differentiation from DNA replication initiation under certain starvation conditions, two otherwise intimately coupled processes. We hypothesize that arresting the developmental program either as motile swarmer cells or as sessile stalked cells improves the chances of survival of C. crescentus during the different starvation conditions.

Influence of light absorption rate on the astaxanthin production by the microalga Haematococcus pluvialis during nitrogen starvation

Large amounts of astaxanthin (about 4% DW) can be produced under nitrogen starvation of Haematococcus pluvialis in photobioreactors (PBRs) exposed to high light conditions to induce a light stress. However, in PBR, the large biomass concentration usually achieved leads to strong light attenuation conditions, which makes complex the analysis of this “light stress”. This study aims to elucidate the role of light transfer in astaxanthin cell content and productivity from the microalga Haematococcus pluvialis during nitrogen starvation. Haematococcus pluvialis was cultivated in a flat-panel PBR in a batch mode with sudden nitrogen starvation conditions and an incident photon flux density (PFD) of 250 µmolhν m−2 s−1. Different initial biomass concentrations ({C}_{{x}_{0}}) were evaluated, 0.21, 0.52, 1.39 and 2.21 kg m−3. As a result, spectral mass absorption cross-sections of Haematococcus pluvialis were measured at different times during nitrogen starvation, and were used to relate the mean rate of photon absorption (MRPA) to the astaxanthin productivity. A minimum initial MRPA of 7000 ± 500 µmolhν kgx−1 s−1 was found necessary to trigger large accumulation of astaxanthin in Haematococcus pluvialis cells (up to 3.21% DW) during nitrogen starvation conditions. The results also demonstrated the link between the MRPA and the daily astaxanthin productivity of Haematococcus pluvialis cultures, introducing then the MRPA as a physical quantity of interest for a rational optimization of the light culture conditions in PBRs.Graphical

Complete Loss ofRelA andSpoT Homologs inArabidopsis Reveals the Importance ofthe Plastidial Stringent Response inthe Interplay between Chloroplast Metabolism andPlant Defense Response.

The highly phosphorylated nucleotide, guanosine tetraphosphate (ppGpp), functions as a secondary messenger in bacteria and chloroplasts. The accumulation of ppGpp alters plastidial gene expression and metabolism, which are required for proper photosynthetic regulation and robust plant growth. However, because four plastid-localized ppGpp synthases/hydrolases function redundantly, the impact of the loss of ppGpp-dependent stringent response on plant physiology remains unclear. We used CRISPR/Cas9 technology to generate an Arabidopsis thaliana mutant lacking all four ppGpp synthases/hydrolases and characterized its phenotype. The mutant showed over 20-fold less ppGpp levels than the wild type under normal growth conditions and exhibited leaf chlorosis and increased expression of defense-related genes as well as salicylic acid and jasmonate levels upon transition to nitrogen-starvation conditions. These results demonstrate that proper levels of ppGpp in plastids are required for controlling not only plastid metabolism but also phytohormone signaling, which is essential for plant defense.

Effects of nitrogen starvation on TAG biosynthesis genes expression in Chlorella vulgaris

Microalgal oils and lipids are the potential sources of sustainable industrial products for pharmaceuticals, nutraceuticals, and cosmeceuticals. However, the production cost of microalgal-based products is still expensive and hinders their marketability. Therefore, research has been focusing on increasing microalgae's oil and lipids content to be economically reasonable. Nutrient stresses were often used to enhance oils and lipids production in microalgae. In this study, the microalga Chlorella vulgaris was exposed to nitrogen starvation for 21 days to examine the effect of nitrogen removal on cell density, total oil content, fatty acids composition, and the expression of two triacylglycerols (TAG) biosynthetic genes, glycerol-3-phosphate acyltransferase (GPAT) and diacylglycerol acyltransferase (DGAT). The results showed that the C. vulgaris cells grew extremely slowly (p<0.05) under nitrogen starvation conditions. However, despite the growth outcome, the nitrogen starvation increased (p<0.05) total oil contents on days 3, 11 and 14 relatives to control. Nitrogen starvation also stimulated (p<0.05) the production of saturated fatty acids (SFA). The primary fatty acids detected were C16:0, C18:0, C18:1, C18:2, C18:3 and C20:0. The effect of nitrogen starvation on the expression of TAG biosynthetic genes, GPAT and DGAT genes were enumerated using real-time PCR. Both GPAT and DGAT were downregulated in this study. Interestingly, the buildup of C18:2 and C18:3 was positively linked with GPAT expression, demonstrating that GPAT affected the synthesis of polyunsaturated fatty acids (PUFA) in nitrogen-starved circumstances. This suggests that the GPAT gene may be altered to increase PUFA in microalgae, notably C18:2 and C18:3.

Heterologous expression of a yeast lipid transporter promotes accumulation of storage compounds in Chlamydomonas

Microalgae offer renewable and sustainable sources for the bioindustrial production of next-generation biofuels and biomaterials. In this work, we heterologously expressed a lipid transporter gene (MMF1) from the major facilitator superfamily in the model green microalga Chlamydomonas reinhardtii, under the influence of a strong light and temperature-inducible promoter. The full-length cDNA sequence of MMF1 from Moesziomyces antarcticus T-34 (MaMMF1) encodes for a 542 amino acid polypeptide with transmembrane efflux pump activity. M. antarcticus is a fungus that can produce and secrete large amounts of glycolipid biosurfactants known as mannosylerythritol lipids (MELs) under nitrogen starvation conditions. Heterologous expression of MaMMF1 in C. reinhardtii resulted in a slight increase in cell biomass, with mutants exhibiting on average about 1.8-fold larger cell sizes. Total lipid content almost doubled in cells expressing MaMMF1 relative to the control strain when cultures were grown for two days under nitrogen starvation. Moreover, average starch content was over 2-fold higher in the mutants. Our results showed that algal strains engineered with the MaMMF1 MEL transporter were capable of possibly redirecting the carbon flux, which resulted in improved growth and accumulation of high-value storage compounds.

A new bacterial endosymbiotic relationship in Kluyveromyces marxianus isolated from the mezcal fermentation process

In Mexico, agave has a significant cultural and economic impact via its use as a raw material for producing fermented drinks. The agave plant grows in arid and nitrogen-deficient soil where it associates with fungi and bacteria to meet nutrient demands, particularly under extreme conditions of fungal growth; this community of microorganisms is called the microbiome. In the microbiome, aromatic volatile synthesis activates the defense response, resistance to abiotic stress, and plant growth. These aromatic volatiles also impact the organoleptic characteristics of fermented beverages and are produced by microorganisms during agave fermentation. This research presents the behavior of K. marxianus cultivated in the presence of extreme and deficient nitrogen concentrations. In addition, a new biological relationship between K. marxianus ITD0090 from mezcal fermentation and Bacillus tequilensis ITD-UANL-01 is reported, confirmed by fluorescence microscopy and intracellular recovery. Our results also reveal that B. tequilensis ITD-UANL-01 is a nitrogen-fixing bacterium that provides assimilable nitrogen to K. marxianus under nitrogen starvation conditions. This phenomenon allows for 144 h of viability without nitrogen. Furthermore, this is the first report of an endosymbiont of K. marxianus ITD0090 in fermentation.

Efficient production of a polyhydroxyalkanoate by Azotobacter vinelandii OP using apple residues as promising feedstock

Fruit residues are attractive substrates for the production of bacterial polyhydroxyalkanoates due to the high contents of fermentable sugars and the fast, simple, and efficient pretreatment methods required. In this study, apple residues, mainly apple peel, were used as the sole carbon source in cultures of the bacterium Azotobacter vinelandii OP to produce poly-3-hydroxybutyrate (P3HB). Conversion from the residue to total sugars was highly effective, achieving conversions of up to 65.4 % w w−1 when using 1 % v v−1 sulfuric acid and 58.3 % w w−1 in the absence of acid (only water). The cultures were evaluated at the shake-flask scale and in 3-L bioreactors using a defined medium under nitrogen starvation conditions. The results showed the production of up to 3.94 g L−1 P3HB in a bioreactor, reaching an accumulation of 67.3 % w w−1 when using apple residues. For the PHB obtained from the cultures with apple residues, a melting point of 179.99 °C and a maximum degradation temperature of 274.64 °C were calculated. A P3HB production strategy is shown using easily hydrolysable fruit residues to achieve production yields comparable to those obtained with pure sugars under similar cultivation conditions.

Expression profile of genes associated with the accumulation of triacylglycerol (TAG) in Auxenochlorella protothecoides KP7 under alkaline pH and nitrogen starvation conditions

Identification of target genes related to lipid accumulation in microalgae can increase triacylglycerol production without affecting biomass production through genetic manipulation. This study is part of our previous study and aimed to determine the effects of nitrogen starvation, alkaline pH (pH 10), and a combination of these factors on the transcriptional profiles of eight genes potentially related to triacylglycerol accumulation in Auxenochlorella protothecoides KP7. As shown previously, triacylglycerol content in Auxenochlorella protothecoides KP7 reached the maximum level under combined stress conditions. In this study, the highest upregulation for the gene encoding phosphoenolpyruvate carboxylase (PEPC) was observed under both nitrogen starvation and combined stress. Nitrogen starvation alone resulted in a significant increase in the expression level of all genes except NADP-malic enzyme (MEchl), which is localized in the chloroplast. Compared with nitrogen starvation alone, the upregulation of pyruvate dehydrogenase complex E2 component (PDHC-E2), glycerol-3-phosphate dehydrogenase (GPDH), diacylglycerol acyltransferase 1 (DGAT1), and MEchl genes further increased under the combined stress. These results suggest that genes encoding the enzymes DGAT1, GPDH, ME, PEPC, and PDHC-E2 are associated with triacylglycerol accumulation and could be used as target genes for genetic modification of Auxenochlorella protothecoides and possibly other microalgal species.

Identification and Functional Analysis of Acyl-Acyl Carrier Protein Δ9 Desaturase from Nannochloropsis oceanica.

The oleaginous marine microalga Nannochloropsis oceanica strain IMET1 has attracted increasing attention as a promising photosynthetic cell factory due to its unique excellent capacity to accumulate large amounts of triacylglycerols and eicosapentaenoic acid. To complete the genomic annotation for genes in the fatty acid biosynthesis pathway of N. oceanica, we conducted the present study to identify a novel candidate gene encoding the archetypical chloroplast stromal acyl-acyl carrier protein Δ9 desaturase. The full-length cDNA was generated using rapid-amplification of cDNA ends, and the structure of the coding region interrupted by four introns was determined. The RT-qPCR results demonstrated the upregulated transcriptional abundance of this gene under nitrogen starvation condition. Fluorescence localization studies using EGFP-fused protein revealed that the translated protein was localized in chloroplast stroma. The catalytic activity of the translated protein was characterized by inducible expression in Escherichia coli and a mutant yeast strain BY4389, indicating its potential desaturated capacity for palmitoyl-ACP (C16:0-ACP) and stearoyl-ACP (C18:0-ACP). Further functional complementation assay using BY4839 on plate demonstrated that the expressed enzyme restored the biosynthesis of oleic acid. These results support the desaturated activity of the expressed protein in chloroplast stroma to fulfill the biosynthesis and accumulation of monounsaturated fatty acids in N. oceanica strain IMET1.

The growth and lipid accumulation of Scenedesmus quadricauda under nitrogen starvation stress during xylose mixotrophic/heterotrophic cultivation.

In order to conquer the block of high cost and low yields which limit to realize the commercialization of microalgal biodiesel, the mixotrophic and heterotrophic cultivation of Scenedesmus quadricauda FACHB-1297 fed on xylose was separately studied employing six forms of media: phosphorus sufficient, phosphorus restricted, and phosphorus starvation were combined with nitrogen sufficient and nitrogen starvation conditions. The maximum lipid content (about 41% of dry weight) was obtained on the 5th day (heterotrophic cultivation) and 8th day (mixotrophic cultivation) under the nitrogen starved and phosphorus sufficient (N0&P) conditions, which was about twofold in comparison to the final lipid content on the sufficient nitrogen condition (control). Under mixotrophic and heterotrophic modes, the highest lipid production was achieved in the N0&P trial, with the value of 274.96mg/L and 193.77mg/L, respectively. Xylose utilization rate of 30-96% under heterotrophic modes was apparently higher than that of 20-50% in mixotrophic modes. In contrast, phosphorus uptake rate of 100% under mixotrophic cultivation was significantly more than that of 60-90% in heterotrophic cultivation. Furthermore, under the condition of heterotrophic cultivation using xylose as a carbon source, the phosphorus had a positive impact on microalgae cell synthesis and the lipid content enhanced with the augmentation in phosphorus concentrations. We suggested that sufficient phosphorus should be supplied for obtaining higher microalgal lipid production in the lack of nitrogen under xylose heterotrophic/mixotrophic condition. This was a highly effective way to obtain efficient microalgae lipid production.

Iron–Sulfur Clusters toward Stresses: Implication for Understanding and Fighting Tuberculosis

Tuberculosis (TB) remains the leading cause of death due to a single pathogen, accounting for 1.5 million deaths annually on the global level. Mycobacterium tuberculosis, the causative agent of TB, is persistently exposed to stresses such as reactive oxygen species (ROS), reactive nitrogen species (RNS), acidic conditions, starvation, and hypoxic conditions, all contributing toward inhibiting bacterial proliferation and survival. Iron–sulfur (Fe-S) clusters, which are among the most ancient protein prosthetic groups, are good targets for ROS and RNS, and are susceptible to Fe starvation. Mtb holds Fe-S containing proteins involved in essential biological process for Mtb. Fe-S cluster assembly is achieved via complex protein machineries. Many organisms contain several Fe-S assembly systems, while the SUF system is the only one in some pathogens such as Mtb. The essentiality of the SUF machinery and its functionality under the stress conditions encountered by Mtb underlines how it constitutes an attractive target for the development of novel anti-TB.

THE SAGA COMPLEX IS ESSENTIAL FOR THE REGULATION OF GENES INVOLVED IN YEAST TREHALOSE METABOLISM

Saccharomyces cerevisiae accumulates trehalose as a stress metabolite in adverse environmental conditions. The trehalose synthesis and breakdown are important for the regulation of trehalose levels within the yeast cell. Therefore, TPS1 and NTH1 gene expressions are tightly regulated during transcription and also translation. Since both genes contain Stress Response Elements (STRE) in the promoter regions, they are co-activated under stress conditions. However, the presence of similar regulatory elements in the promoter of both genes shows that these genes undergo a different regulation at the transcriptional level. In our study, the role of the Spt-Ada-Gcn5 Acetyltransferase (SAGA) complex in the transcriptional regulation of TPS1 and NTH1 genes was determined in nutrient-poor environment. For that purpose, the wild type and Δada1 mutant yeast cells, where Ada1p is a member of the SAGA complex, were grown in normal and nitrogen starvation conditions. In addition, trehalose level was detected enzymatically in both wild type and mutant yeast cells. In silico promoter analysis of TPS1 and NTH1 promoters revealed that the STRE sequences required for binding of Msn2/4 transcription factors are closed by nucleosomes at the NTH1 promoter, but open at the TPS1 promoter. In the absence of Ada1p, stress-induced promoter activation in the TPS1 gene was observed, while NTH1 gene expression was not activated. According to these results, the nucleosomes spanning the STRE sequences could not be mobilized in the absence of Ada1 protein, and therefore the Msn2/4 transcription factors cannot bind to the promoter and activate the NTH1 gene expression under stress conditions. It was also observed that in the absence of Ada1p, trehalose accumulation was reduced regardless of stress conditions.

Nitrogen Starvation Enhances the Production of Saturated and Unsaturated Fatty Acids in Aurantiochytrium sp. PKU#SW8 by Regulating Key Biosynthetic Genes.

Nitrogen deprivation is known to improve lipid accumulation in microalgae and thraustochytrids. However, the patterns of fatty acid production and the molecular mechanisms underlying the accumulation of unsaturated and saturated fatty acids (SFAs) under nitrogen starvation remain largely unknown for thraustochytrids. In this study, batch culture experiments under nitrogen replete and nitrogen starvation conditions were performed, and the changes in the transcriptome of Aurantiochytrium sp. PKU#SW8 strain between these conditions were investigated. Our results showed improved yields of total fatty acids (TFAs), total unsaturated fatty acids, and total SFAs under nitrogen starvation, which suggested that nitrogen starvation favors the accumulation of both unsaturated and saturated fatty acids in PKU#SW8. However, nitrogen starvation resulted in a more than 2.36-fold increase of SFAs whereas a 1.7-fold increase of unsaturated fatty acids was observed, indicating a disproportionate increase in these groups of fatty acids. The fabD and enoyl-CoA hydratase genes were significantly upregulated under nitrogen starvation, supporting the observed increase in the yield of TFAs from 2.63 ± 0.22 g/L to 3.64 ± 0.16 g/L. Furthermore, the pfaB gene involved in the polyketide synthase (PKS) pathway was significantly upregulated under nitrogen starvation. This suggested that the increased expression of the pfaB gene under nitrogen starvation may be one of the explanations for the increased yield of docosahexaenoic acid by 1.58-fold. Overall, our study advances the current understanding of the molecular mechanisms that underlie the response of thraustochytrids to nitrogen deprivation and their fatty acid biosynthesis.

Expression levels of nitrogen assimilation-related genes, physiological responses, and morphological adaptations of three indica rice (Oryza sativa L. ssp. indica) genotypes subjected to nitrogen starvation conditions.

Nitrogen (N) is an essential nutrient available to the plants in form of nitrate and ammonium. It is a macronutrient important for the plant growth and development, especially in cereal crops, which consume it for the production of amino acids, proteins/enzymes, nucleic acids, cell wall complexes, plant hormones, and vitamins. In rice production, 17kgN uptake is required to produce 1 ton of rice. Considering this, many techniques have been developed to evaluate leaf greenness or SPAD value for assessing the amount of N application in the rice cultivar to maximize the grain yield. The aim of the present study was to investigate the morpho-physiological characteristics and relative expression level of N assimilation in three different rice genotypes (MT2, RD31, KDML105) under 1.00 × (full N), 0.50 × , 0.25 × (N depletion), and 0.00 × (N deficiency) at seedling stage and the morpho-physiological traits and the grain yield attributes under 1.00 × (full N) and 0.25 × (N depletion) were compared. Leaf chlorosis and growth inhibition in rice seedlings under N deficiency were evidently observed. Shoot height, number of leaves, shoot fresh weight, shoot dry weight, and root fresh weight in KDML105 under N deficiency were decreased by 27.65%, 42.11%, 65.44%, 47.90%, and 54.09% over the control (full N). Likewise, leaf greenness was lowest in KDML105 under N deficiency (78.57% reduction over the full N), leading to low photosynthetic abilities. In addition, expression of nitrogen assimilation-related genes, OsNR1, OsGln1;1, and OsGln2, in KDML105 under N depletion were increased within 3h and then declined after the long incubation period, whereas those were unchanged in cvs. MT2 and RD31. Similarly, relative expression level of OsNADH-GOGAT, OsFd-GOGAT, and OsAspAt1 in KDML105 was peaked when subjected to 0.50 × N for 6h and then declined after the long incubation period. Moreover, overall growth characters and physiological changes in cv. RD31 at vegetative stage under 0.25 × N were retained better than those in cvs. KDML105 and MT2, resulting in high yield at the harvesting process. In summary, N assimilated-related genes in rice seedlings under N depletion were rapidly regulated within 3-6h, especially cv. KDML105 and MT2, then downregulated, resulting in physiological changes, growth inhibition, and yield reduction.

Inhibition of monogalactosyldiacylglycerol synthesis by down-regulation of MGD1 leads to membrane lipid remodeling and enhanced triacylglycerol biosynthesis in Chlamydomonas reinhardtii

BackgroundMembrane lipid remodeling involves regulating the physiochemical modification of cellular membranes against abiotic stress or senescence, and it could be a trigger to increase neutral lipid content. In algae and higher plants, monogalactosyldiacylglycerol (MGDG) constitutes the highest proportion of total membrane lipids and is highly reduced as part of the membrane lipid remodeling response under several abiotic stresses. However, genetic regulation of MGDG synthesis and its influence on lipid synthesis has not been studied in microalgae. For development of an industrial microalgae strain showing high accumulation of triacylglycerol (TAG) by promoting membrane lipid remodeling, MGDG synthase 1 (MGD1) down-regulated mutant of Chlamydomonas reinhardtii (Cr-mgd1) was generated and evaluated for its suitability for biodiesel feedstock.ResultsThe Cr-mgd1 showed a 65% decrease in CrMGD1 gene expression level, 22% reduction in MGDG content, and 1.39 and 5.40 times increase in diacylglyceryltrimethylhomoserines (DGTS) and TAG, respectively. The expression levels of most genes related to the decomposition of MGDG (plastid galactoglycerolipid degradation1) and TAG metabolism (diacylglycerol O-acyltransferase1, phospholipid:diacylglycerol acyltransferase, and major lipid droplet protein) were increased. The imbalance of DGDG/MGDG ratio in Cr-mgd1 caused reduced photosynthetic electron transport, resulting in less light energy utilization and increased reactive oxygen species levels. In addition, endoplasmic reticulum stress was induced by increased DGTS levels. Thus, accelerated TAG accumulation in Cr-mgd1 was stimulated by increased cellular stress as well as lipid remodeling. Under high light (HL) intensity (400 µmol photons/m2/s), TAG productivity in Cr-mgd1–HL (1.99 mg/L/d) was 2.71 times higher than that in wild type (WT–HL). Moreover, under both nitrogen starvation and high light intensity, the lipid (124.55 mg/L/d), TAG (20.03 mg/L/d), and maximum neutral lipid (56.13 mg/L/d) productivity were the highest.ConclusionsBy inducing lipid remodeling through the mgd1 gene expression regulation, the mutant not only showed high neutral lipid content but also reached the maximum neutral lipid productivity through cultivation under high light and nitrogen starvation conditions, thereby possessing improved biomass properties that are the most suitable for high quality biodiesel production. Thus, this mutant may help understand the role of MGD1 in lipid synthesis in Chlamydomonas and may be used to produce high amounts of TAG.

Introgression of a novel gene AhBINR differentially expressed during PGPR Brachybacterium saurashtrense-Arachis hypogaea interaction enhances plant performance under nitrogen starvation and salt stress in tobacco

Plant growth-promoting rhizobacteria provide endurance during environmental stress conditions. Previously, we have shown that the interaction of the halotolerant diazotrophic bacteria Brachybacterium saurashtrense JG06 induces physio-biochemical and molecular changes in Arachis hypogaea under nitrogen starvation conditions. Here we deciphered the role of a novel gene AhBINR that was differentially overexpressed in A. hypogaea after interaction with B. saurashtrense JG06 under nitrogen deficit conditions. Overexpression of the AhBINR gene in the model plant (tobacco) showed higher growth parameters (root length, shoot length, fresh weight, and dry weight) under nitrogen starvation and salt stress in comparison to the wild type and vector control. Transgenic plants were enabled with a higher photosynthesis rate, which provides the support for better performance under N2 starvation and salt stress. Results showed that the transgenic plants overexpressing the AhBINR gene had better physiological status and lower ROS accumulation under adverse conditions. Microarray transcriptome analysis showed that the transcription factors, biotic and abiotic stress, photosynthesis, and metabolism-related genes were differentially expressed (total 736 and 6530 genes were expressed under nitrogen deficit and salt stress conditions, respectively at a 5-fold change level) in comparison to wild type plants. Overall results showed the involvement of the AhBINR gene in the activation of the abiotic stress (nitrogen starvation and salt stress) related pathways, which can be overexpressed after legume-rhizobacterial interaction.