Effects Of Nitrogen Starvation Research Articles

Effect of alkaline pH and nitrogen starvation on the triacylglycerol (TAG) content, growth, biochemical composition, and fatty acid profile of Auxenochlorella protothecoides KP7

Microalgae can accumulate substantial amounts of triacylglycerol, the primary feedstock for biodiesel production, under different stress conditions. In this study, exposure to alkaline pH (pH 10) using carbonate-bicarbonate buffer, nitrogen starvation, and both were applied to induce triacylglycerol accumulation in Auxenochlorella protothecoides KP7. In addition to triacylglycerol accumulation, growth, biochemical composition, and fatty acid profiles were examined in detail and compared. Our results illustrated that the combination of nitrogen starvation and alkaline pH led to significant increases in triacylglycerol and starch contents and drastic decreases in chlorophyll and protein contents compared with their control levels. Triacylglycerol levels, the triacylglycerol/total lipid ratio, and lipid productivities were significantly increased after 7 days of cultivation under the combined stress compared with the effects of nitrogen starvation alone. After prolonged cultivation under the combined stress, triacylglycerol content reached up to 25.01% of the dry cell weight and constituted up to 81.25% of total lipids. Under the combined stress, the major fatty acids in the total lipid of A. protothecoides KP7 were C18:1 (57.5%), C18:2 (19.5%), C16:0 (12.5%), and C18:3 (5.6%), and the percentages of saturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids represented 16.3, 58.4, and 25.1% of the total fatty acids, respectively. Furthermore, the combined stress was associated with the lowest iodine value (97.64 g I2 (100 g)−1), highest oxidative stability (7.29 h), and highest cetane number (55.26) among the examined conditions. The use of nitrogen-free medium containing carbonate-bicarbonate buffer for cultivation appears promising for enhancing triacylglycerol production in A. protothecoides.

Effects of nitrogen starvation on protein and carbohydrate contents of some marine microalgae and their efficiency as food for the rotifer Brachionus plicatilis (Müller)

Effects of nitrogen starvation on protein and carbohydrate contents of some marine microalgae and their efficiency as food for the rotifer Brachionus plicatilis (Müller)

Physiological plasticity of symbiotic Desmodesmus (Chlorophyceae) isolated from taxonomically distant white sea invertibrates

The physiological heterogeneity of closely related symbiotic microalgae from taxonomically distant animal hosts was studied for the first time. Three strains of unicellular algae from the genus Desmodesmus (Chlorophyceae) isolated from White Sea benthic invertebrates were used as the object in this study. The effects of nitrogen starvation and high light intensity on the growth, changes in chlorophyll and total carotenoid contents and fatty acid content and composition of the microalgal cell lipids were followed. Nitrogen starvation declined the biomass accumulation rate as well as chlorophyll and carotenoid contents on the background of the enhanced fatty acid accumulation in all strains studied. The ultrastructural study revealed the reduction of photosynthetic apparatus and an increase in the proportion of cell volume occupied by oil bodies and starch grains as well as an increase in the cell wall thickness. A decline in the effective per cell irradiance in the cultures of higher cell density, as a rule, slowed down the changes in pigment and fatty acids composition characteristic of nitrogen starvation. The rates of biomass accumulation and cell biochemical composition under the nitrogen-starvation conditions were strain-specific. In most cases, the decline in chlorophylls proceeded at a higher rate in comparison with that of carotenoid decline. In two of the three strains studied, both these process occurred synchronously with the decline in the unsaturation of the cell lipid fatty acids. The possibilities of biotechnological application of the symbiotic microalgae are discussed with the peculiarities of their stress physiology in mind.

Effects of nitrogen starvation on the photosynthetic physiology of a tropical marine microalga Rhodomonas sp. (Cryptophyceae)

The effects of nitrogen starvation on biomass composition and photosynthetic function were examined in the marine cryptophyte Rhodomonas sp. Batch-cultured cells in N-sufficient medium showed a 2.5-fold increase in total carbohydrate content, and a 33% increase in cell volume when the cultures reached the stationary growth phase. These cultures also increased the ratio of phycoerythrin (PE)/hydrosoluble proteins from 6 to 22% by the 4th and 10th day of culture, respectively. In contrast, light-saturated photosynthetic activity ( P m) progressively decreased, and the value obtained at the beginning of the stationary phase was about 45% of that obtained for cells in the late exponential growth phase. Transfer to N-lacking medium caused a 3.2-fold increase in cell volume. N starvation also triggered a rapid decline in N-containing compounds such as hydrosoluble proteins and photosynthetic pigments, causing an almost complete loss of PE. The ratio of PE/hydrosoluble proteins decreased from 6 to 1% after 6 d of N deprivation. Furthermore, the PSII fluorescence capacity declined under N-starved conditions, which caused a pronounced decrease in both the P m (circa 90%) and the apparent photosynthetic efficiency (circa 55%). Under these conditions, photosynthetically fixed carbon was used to synthesize large amounts of carbohydrates. We suggest that, in addition to the role of phycoerythrin as a light-harvesting pigment, Rhodomonas sp. responds to N-depleted conditions by mobilizing combined nitrogen from biliproteins.

The effect of nitrogen starvation on the ultrastructure and pigment composition of chloroplasts in the acidothermophilic microalga Galdieria sulphuraria

We studied the effect of nitrogen starvation on growth indices, vitality, ultrastructure, and the photosynthetic apparatus of unique acidothermophilic microalga Galdieria sulphuraria (Galdieri) Merola. Long-term nitrogen starvation ceased G. sulphuraria growth and cell division. During the first days of starvation, phycobiliproteins degraded first, then the content of chlorophyll and carotenoids decreased to trace amounts, chloroplast reduced, cell wall became thinner, and storage compounds accumulated. However, the cells were alive. A comparison with the effects of nitrogen starvation on other photosynthesizing organisms showed that suppression of cell division, reduction of the photosynthetic apparatus to some minimum, and accumulation of storage compounds are a universal response to this stress.

CHANGES IN CELL COMPOSITION AND LIPID METABOLISM MEDIATED BY SODIUM AND NITROGEN AVAILABILITY IN THE MARINE DIATOM PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

ABSTRACTThe effects of nitrogen starvation in the presence or absence of sodium in the culture medium were monitored in batch cultures of the marine diatom Phaeodactylum tricornutum Bohlin. During nitrogen starvation in the presence of sodium, cell nitrogen and chlorophyll a decreased, mainly as a consequence of continued cell division. These decreases were accompanied by decreases in the rates of photosynthesis and respiration. There was no change in either cell volume or carbohydrate, but both carbon and lipid increased. During nitrogen starvation in the absence of sodium, cell division ceased. Cell nitrogen and chlorophyll a remained constant, and respiration did not decrease, but the changes in the photosynthetic rate and the lipid content per cell were similar to cultures that were nitrogen‐starved in the presence of sodium. The carbon‐to‐nitrogen ratio increased in both cultures. Nitrogen, in the form of nitrate, and sodium were resupplied to cultures that had been preconditioned in nitrogen‐ and sodium‐deficient medium for 5 d. Control cultures to which neither nitrate or sodium were added remained in a static state with respect to cell number, volume, and carbohydrate but showed slight increases in lipid. Cells in cultures to which 10 mM nitrate alone was added showed a similar response to cultures where no additions were made. Cells in cultures to which 50 mM sodium alone was added divided for 2 d, with concomitant small decreases in all measured constituents. Cell division resumed in cultures to which both sodium and nitrate were added. The lipid content fell dramatically in these cells and was correlated to metabolic oxidation via measured increases in the activity of the glyoxylate cycle enzyme, isocitrate lyase. We conclude that lipids are stored as a function of decreased growth rate and are metabolized to a small extent when cell division resumes. However, much higher rates of metabolism occur if cell division resumes in the presence of a nitrogen source.

New Elements in the Mitotic Control of the Fission Yeast Schizosaccharomyces pombe

The p107wee1 protein kinase plays a central role in regulating the cell cycle of fission yeast. It mediates transmission of signal(s) related to the nutritional status of the cell to the p34cdc2 protein kinase, which is an active component of the MPF complex driving cells into mitosis. p107wee1 is itself subject to control by the products of other genes such as nim1+/cdr1+, win1+, and perhaps wis1+ and other wis+ genes. At present, the relationships between these genes and their possible roles in the mitotic control are unclear and must await further analysis (Fig. 5). It is likely that some of the gene products are concerned with the sensing and/or transmission of nutritional signals. p107wee1 negatively regulates the activity of p34cdc2, probably by direct tyrosine phosphorylation, and also appears to regulate the activities of the cdc1+ and cdc27+ gene products. The effects of nitrogen starvation and of wee1 mutations on conditional lethal mutations at the cdc1, cdc2, and cdc27 loci, taken together, support the largely speculative model shown in Figure 5. During the normal cycle, the balance between phosphorylated and dephosphorylated p34cdc2 changes such that at the appropriate time, p34cdc2 is activated and the cell enters mitosis. We suggest that the cdc1+ and cdc27+ products may be regulated in a similar way. Such a mechanism would ensure coordinated activation of these and perhaps other proteins required for the G2/M transition. There are, of course, many uncertainties, and these must await elucidation by biochemical and genetic analysis.

Effects of nitrogen starvation on the function and organization of the photosynthetic membranes in Cryptomonas maculata (Cryptophyceae)

Nitrogen deficiency affects both photosystems and the antennae pigment systems in the photosynthetic apparatus of the marine alga, Cryptomonas maculata. Under increasing energy fluence rates, O2 evolution in nitrogen-deficient (-N) cell suspensions never reached a positive value; in control cultures (+N), O2 evolution increased and was saturated at about 6.4 W·m(-2) with about 100 μmol O2·mg chlorophyll(-1)·h(-1). During fluorescence-induction experiments at room temperature, Fo and Fmax were significantly increased in-N cells whereas the Fvar/Fmax ratio decreased from 0.6 to 0.1. These observations can be correlated with a significantly decreased population of 12.5-nm-size particles in the exoplasmic-fracture (EF) faces of freeze-cleaved thylakoid membranes in-N cells (Rhiel et al., 1985, Protoplasma 129, 62-73). The EF particles are suggested to represent photosystem II associated with chlorophyll a/c-protein complexes (LHCP). The banding pattern of isolated and Triton X-100-solubilized thylakoid membranes of both +N and-N cells in sucrose gradients showed that the LHCP is still present in-N cells. The same applies to sodium dodecyl sulfate-polyacrylamide gel electrophoresis of these membrane fractions. The reduced number of the 12.5-nm particles in the EF faces of-N cells may be a result of decoupling of the LHCP constituents of the photosystem-II complex rather than their degradation. This is supported by high values for the initial fluorescence Fo in fluorescence-induction experiments and, in part, is indicated by the shift of the maximal fluorescence emission from 693 nm in +N to 684 nm in-N cells. The lack of the CP1 band in the gels of sodium dodecyl sulfate-solubilized thylakoid membranes from-N cells after electrophoresis demonstrates that photosystem I is also severely affected.

Effect of nitrogen starvation on the morphology and ultrastructure of the cyanobacterium Mastigocladus laminosus

The effects of nitrogen starvation on the morphology and ultrastructure of the branching, filamentous cyanobacterium Mastigocladus laminosus were examined with light and electron microscopy. The internal ultrastructural characteristics of vegetative cells changed markedly during nitrogen starvation. Carboxysomes were degraded, while polyphosphate bodies and lipid bodies accumulated. The ultrastructure of mature heterocysts was also affected by nitrogen starvation; their intracytoplasmic membranes vesiculated to form vacuolelike structures and, eventually, large empty regions in the cytoplasm. Nitrogen starvation stimulated extensive heterocyst differentiation in M. laminosus, producing heterocyst frequencies of 17.5% in narrow filaments and 28.3% in wide filaments within 44 h after transfer to N-free conditions. Cells in wide filaments differentiated so extensively that only 16.8% of them failed to initiate the differentiation process within 44 h.

The effects of nitrogen starvation on the concentration of free amino acids in Myzus persicae (Sulzer) (Homoptera, Aphidae)

Amino acid analyses performed on aphids taken directly from their host, and on aphids which had been nitrogen-starved for 24 and 48 hr, indicated that as the starvation period increased, the concentration of sixteen free amino acids decreases. Final concentrations were 50–70 per cent of the original values. Cystine increased 1100 per cent. In addition, twenty-eight of thirty-one unidentified ninhydrin-positive compounds increased in concentration as the starvation period continued. The aphids used in these experiments had been artificially fed on 18% sucrose solution.

The formation of 4-hydroxyphenylpyruvic acid and phenylpyruvic acid by tryptophan auxotrophs and wild-type Aerobacter aerogenes considered in relation to the general aromatic pathway

4-Hydroxyphenylpyruvic acid and phenylpyruvic acid are accumulated by cell suspensions of tryptophan auxotrophs and a wild-type strain of Aerobacter aerogenes when these are transferred from conditions of adequate nitrogen to conditions of nitrogen starvation. A mutant strain which requires the full supplement of five aromatic compounds does not form hydroxyphenylpyruvic acid or phenylpyruvic acid. The compounds 4-hydroxybenzaldehyde and 3,4-dihydroxybenzoic acid may also be detected in supernatants after the cells are incubated with substrates lacking nitrogen. The effects of nitrogen starvation are discussed in relation to the critical specific steps of tryptophan biosynthesis and the problem of the branch point in aromatic biosynthesis. It is suggested that nitrogen is introduced in tryptophan biosynthesis at the first step specific to this pathway.