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Effects of nitrogen allocation and photosynthetic proteins response in peanut leaves on photosynthesis under conditions of water scarcity and nitrogen deficiency.

Leaf nitrogen allocation and photosynthetic proteins response can affect net photosynthetic rate (Pn), ultimately influencing crop yield under diverse environmental stresses. However, the internal relationship between Pn with leaf nitrogen allocation and photosynthetic proteins response under nitrogen or water scarcity in peanut (Arachis hypogaea L.) remains elusive. Here, comprehensive physiological property and proteomic analyses of peanut were conducted, revealing that both nitrogen and water scarcity remarkably impeded leaf growth and reduced Pn. Nitrogen deficiency significantly reduced the total nitrogen content per unit leaf area (Narea), chlorophyll content, and Pn, whereas drought stress caused a greater decline in photosynthetic nitrogen use efficiency (PNUE). The allocation of leaf nitrogen to photosynthetic components, including the carboxylation system and electron transport system in leaves, was significantly reduced when subjected to individual or combined deficiency. Proteomic analyses exhibited that several key photosynthetic proteins underwent a decrease under both single and combined water and nitrogen deficiency conditions. Thereby, Pn may decline due to the disruption of nitrogen allocation and down-regulated expression of photosynthetic proteins under these stress conditions. Our findings establish a benchmark for future research exploring the roles of leaf nitrogen allocation and photosynthetic proteins in the plant's response to nitrogen or water deficiency.

Nitrogen↗

Regulation of photosynthesis in nitrogen deficient wheat seedlings.

Nitrogen effects on the regulation of photosynthesis in wheat (Triticum aestivum L., cv Remia) seedlings were examined. Ribulose 1,5-bisphosphate carboxylase/oxygenase was rapidly extracted and tested for initial activity and for activity after incubation in presence of CO(2) and Mg(2+). Freeze clamped leaf segments were extracted for determinations of foliar steady state levels of ribulose 1,5-bisphosphate, triose phosphate, 3-phosphoglycerate, ATP, and ADP. Nitrogen deficient leaves showed increased ATP/ADP and triose phosphate/3-phosphoglycerate ratios suggesting increased assimilatory power. Ribulose 1,5-bisphosphate levels were decreased due to reduced pentose phosphate reductive cycle activity. Nevertheless, photosynthesis appeared to be limited by ribulose 1,5-bisphosphate carboxylase/oxygenase, independent of nitrogen nutrition. Its degree of activation was increased in nitrogen deficient plants and provided for maximum photosynthesis at decreased enzyme protein levels. It is suggested that ribulose 1,5-bisphosphate carboxylase/oxygenase activity is regulated according to the amount of assimilatory power.

Journal Article↗

Photosynthetic CO(2) assimilation, chlorophyll fluorescence and photoinhibition as affected by nitrogen deficiency in maize plants.

Effects of nitrogen deficiency on photosynthetic CO(2) assimilation, PSII photochemistry and photoinhibition were investigated in maize plants grown under natural illumination. Nitrogen-deficient plants had a significantly smaller CO(2) assimilatory capacity, but they showed little changes in the maximal efficiency of PSII photochemistry, the rate of Q(A) reduction and the heterogeneity of PSII reaction centers, suggesting that nitrogen deficiency had little effects on PSII primary photochemistry and photoinhibition even under natural illumination. However, modifications in PSII photochemistry under the steady state of photosynthesis induced by nitrogen deficiency were observed. This is reflected in decreases in the quantum yield of PSII electron transport, the efficiency of excitation energy capture by open PSII reaction centers, and the photochemical quenching coefficient and an increase in the non-photochemical quenching coefficient. These results suggest that modifications of PSII photochemistry under the steady state of photosynthesis may be a mechanism to downregulate photosynthetic electron transport so that production of ATP and NADPH would be in equilibrium with the decreased demand in the Calvin cycle in nitrogen-deficient plants. On the other hand, the nitrogen-deficient plants still exhibited increased susceptibility to photoinhibition when exposed to very high irradiance, although nitrogen deficiency induced no photoinhibition under natural illumination. Our results suggest that such increased susceptibility to photoinhibition was associated with the increased accumulation of inactivated PSII reaction centers, the decreased capacity of non-photochemical quenching, and the increased fraction of the reduction state of Q(A).

Journal Article↗

Batch biological treatment of nitrogen deficient synthetic wastewater using Azotobacter supplemented activated sludge.

Biological treatment of nitrogen deficient wastewaters are usually accomplished by external addition of nitrogen sources to the wastewater which is an extra cost item. As an alternative for effective biological treatment of nitrogen deficient wastewaters, the nitrogen fixing bacterium, Azotobacter vinelandii, was used in activated sludge and also in pure culture. Total organic carbon (TOC) removal performances of Azotobacter-added and free activated sludge cultures were compared at different initial TN/TOC ratios. The rate and extent of TOC removal were comparable for all cultures when initial TN/TOC ratio was larger than 0.12; however, both the rate and extent of TOC removal from nitrogen deficient (TN/TOC<12%) synthetic wastewater were improved by using Azotobacter-added activated sludge as compared to the Azotobacter-free activated sludge culture. More than 90% TOC removal was obtained with pure Azotobacter or Azotobacter-added activated sludge culture from a nitrogen deficient synthetic wastewater.

Azotobacter↗

Effect of Nitrogen Deficiency upon Translocation of C in Sugarcane.

Withholding nitrogen decreased the percentages of nitrogen and chlorophyll in the blades; reduced the total fixation of radioactive carbon dioxide at 15, 37, and 178 seconds; and changed the relative composition of fixation products. Translocation of radioactive photosynthate from the fed part down the attached blade and into the stalk was less in the plants deprived of nitrogen than in the control plants supplied with nitrogen. Both the percentage of total activity translocated and the velocity of transport were decreased by nitrogen deficiency. During a translocation period of 90 minutes the minus nitrogen blade retained more (14)C-sucrose than the control in the fed part and the blade below the fed part, but it sent less (14)C-sucrose to the sheath of the fed leaf. Thus translocation decreased with nitrogen deficiency not for lack of sucrose but for some other reason. Although withholding nitrogen decreased translocation of labeled carbon in and from attached blades, there was no effect upon transport in detached blades. The effect of nitrogen deficiency upon translocation may be indirect and secondary to the effect upon growth of the plant as a whole.

Journal Article↗

Nitrogen deficiency enhances expression of specific MYB and bHLH transcription factors and accumulation of end products in the flavonoid pathway.

Expression of regulators of the flavonoid pathway was examined in Arabidopsis thaliana wild type and pap1D plants, the latter being a T-DNA activation-tagged line over-expressing the PAP1/MYB75 gene which is a positive regulator of the pathway. Anthocyanin accumulation was induced in plants grown in soil, on agar plates, and hydroponics by withdrawing nitrogen from the growth medium. The agar-grown seedlings and rosette stage plants in hydroponics were further explored, and showed that nitrogen deficiency resulted in the accumulation of not only anthocyanins, but also flavonols. The examination of transcript levels showed that the general flavonoid pathway regulators PAP1 and PAP2 were up-regulated in response to nitrogen deficiency in wild type as well as pap1D plants. Interestingly, PAP2 responded much stronger to nitrogen deficiency than PAP1, 200- and 6-fold increase in transcript levels, respectively, for wild-type seedlings. In rosette leaves the increase was 900-fold for PAP2 and 6-fold for PAP1. At least three different bHLH domain transcription factors promote anthocyanin synthesis, and transcripts for one of these, i.e. GL3 were found to be sixfold enhanced by nitrogen deficiency in rosette leaves. The MYB12 transcription factor, known to regulate flavonol synthesis, was slightly induced by nitrogen deficiency in seedlings. In conclusion, four out of eight regulators involved in the flavonoid pathway showed an enhanced expression from 2 to 1,000 times in response to nitrogen deficiency. Together with MYB factors, especially PAP2, GL3 appears to be the BHLH partner for anthocyanin accumulation in response to nitrogen deficiency.

Agar↗

Chloroplast Fatty Acid transformations in nitrogen-deficient and senescent tissues.

The fatty acids of plastids from several types of mineral-deficient and senescent tissues were analyzed. Incorporation of acetate into long-chain fatty acids of leaf tissue and of plastids from nitrogen-deficient and normal plants was determined. In general, the senescent and nitrogen-deficient chloroplasts contained a higher ratio of saturates to unsaturates than did plastids from younger tissues and from tissues grown on a complete nutrient.Nitrogen-deficient leaf tissue and plastids were capable of rapidly incorporating acetate into some of the fatty acids, especially palmitic and oleic acids. However, the comparative rate of acetate incorporation into linolenic acid in nitrogen-deficient chlorophyllous tissue was less than in tissue grown on a complete nutrient. With the addition of UDP-glucose to a reaction mixture containing added cofactors for noncyclic photosynthetic phosphorylation the relative incorporation of acetate into linolenate as compared to palmitate was increased in both the nitrogen-deficient and normal leaf tissue. This would indicate that nitrogen-deficient tissues have the enzymic systems for forming long-chain fatty acids but that the reduced photosynthesis limits the amount of precursors for the formation of lipids, especially galactolipids. However, nothing is known about the rate of fatty acid degradation under these conditions.

Journal Article↗

Characterization of Synechocystis sp. strain PCC 6803 and deltanbl mutants under nitrogen-deficient conditions.

The impact of nitrogen deficiency on the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 and three nbl (non-bleaching) mutants (deltanblA1, deltanblA2 and deltanblB) was investigated. The deltanblA mutants entered a non-dividing, dormant state soon after the initiation of nitrogen starvation. The cells became larger, the membrane system was disorganized, and ribosomes were found near the membranes much less frequently. Photosystem II (PSII) activity declined to approximately 10% of the wild-type level and the amount of D1 protein declined precipitously, despite adequate psbA transcription; PSI activity declined, but more slowly. Transcription from PSII (except psbA), PSI and phycobilisome genes was very low. Fluorescence at 77K indicated many partially assembled or unassembled phycobilisomes. The level of transcript accumulation increased to normal by 4 h after the readdition of nitrogen to the culture. When NblA was present, the phycobilisomes were degraded to provide a nitrogen source for continued growth and metabolism. An important difference between the wild-type, mutant deltanblB, and the deltanblA mutants was seen in the rod linker proteins. Under nitrogen-deprivation condition, the L(R)33 and L(R)34.5 linker proteins were extensively degraded in the wild-type and deltanblB mutant, but remained intact in the deltanblA mutants.

Bacterial Proteins↗

The regreening of nitrogen-deficient Chlorella fusca II. Structural changes during synchronous regreening.

Chlorella fusca, strain 211-15, cells degreened in a nitrogen-deficient mineral growth medium in the light for 4-6 weeks were regreened for up to 24 hrs in a nitrogen rich medium that leads to synchronous cell division at 24-26 hrs. Structural changes in the plastid membranes during the regreening period were observed by thin section and freeze-fracture electron microscopy. Nitrogen-deficient plastids were found to have non-appressed lamellae, prolamellar body-like membrane aggregations, and only 2 types of freeze-fracture face. At this time no photosynthetic oxygen evolution could be demonstrated. After 6 hrs regreening the plastid lamellae had fused to form bands of appressed lamellae and the four types of freeze-fracture face, described previously, were visible. At this time photosynthetic oxygen evolution could be demonstrated. After 24 hrs regreening the plastids had an appearance typical of normally grown Chlorella and had commenced to divide. Supporting evidence for these developmental stages is presented from isolated chloroplast particle fractions. An unusual type of cell wall proliferation was observed in the nitrogen-deficient Chlorella cells that resulted in the laying down of several walls, each with a trilaminar component.

Cell Division↗

Biomass content governs fermentation rate in nitrogen-deficient wine musts.

Problematic fermentations are common in the wine industry. Assimilable nitrogen deficiency is the most prevalent cause of sluggish fermentations and can reduce fermentation rates significantly. A lack of nitrogen diminishes a yeast's metabolic activity, as well as the biomass yield, although it has not been clear which of these two interdependent factors is more significant in sluggish fermentations. Under winemaking conditions with different initial nitrogen concentrations, metabolic flux analysis was used to isolate the effects. We quantified yeast physiology and identified key metabolic fluxes. We also performed cell concentration experiments to establish how biomass yield affects the fermentation rate. Intracellular analysis showed that trehalose accumulation, which is highly correlated with ethanol production, could be responsible for sustaining cell viability in nitrogen-poor musts independent of the initial assimilable nitrogen content. Other than the higher initial maintenance costs in sluggish fermentations, the main difference between normal and sluggish fermentations was that the metabolic flux distributions in nitrogen-deficient cultures revealed that the specific sugar uptake rate was substantially lower. The results of cell concentration experiments, however, showed that in spite of lower sugar uptake, adding biomass from sluggish cultures not only reduced the time to finish a problematic fermentation but also was less likely to affect the quality of the resulting wine as it did not alter the chemistry of the must.

Biomass↗

Water Relations of Cotton Plants under Nitrogen Deficiency: II. Environmental Interactions on Stomata.

Nitrogen deficiency in cotton plants (Gossypium hirsutum L.) considerably increased the sensitivity of stomata to water stress. At air temperatures of 27, 35, and >/=40 C, threshold potentials for complete stomatal closure were -10, -15, and -26 bars in N-deficient plants and -20, -20, and -30 bars in high-N plants, respectively. This three-way interaction among N supply, water potential, and air temperature was similar to that exerted on leaf expansion. The effects of N supply on stomatal behavior could not be explained on the basis of either osmotic or structural considerations. Rather, effects of N deficiency on mesophyll and stomata were independent and divergent. Stomatal behavior may impart a stress avoidance type of drought resistance to N-deficient plants.

Journal Article↗

Additive contribution of AMT1;1 and AMT1;3 to high-affinity ammonium uptake across the plasma membrane of nitrogen-deficient Arabidopsis roots.

In Arabidopsis four root-expressed AMT genes encode functional ammonium transporters, which raises the question of their role in primary ammonium uptake. After pre-culturing under nitrogen-deficiency conditions, we quantified the influx of (15)N-labeled ammonium in T-DNA insertion lines and observed that the loss of either AMT1;1 or AMT1;3 led to a decrease in the high-affinity ammonium influx of approximately 30%. Under nitrogen-sufficient conditions the ammonium influx was lower in Columbia glabra compared with Wassilewskija (WS), and AMT1;1 did not contribute significantly to the ammonium influx in Col-gl. Ectopic expression of AMT1;3 under the control of a 35S promoter in either of the insertion lines amt1;3-1 or amt1;1-1 increased the ammonium influx above the level of their corresponding wild types. In transgenic lines carrying AMT-promoter-GFP constructs, the promoter activities of AMT1;1 and AMT1;3 were both upregulated under nitrogen-deficiency conditions and were localized to the rhizodermis, including root hairs. AMT gene-GFP fusions that were stably expressed under the control of their own promoters were localized to the plasma membrane. The double insertion line amt1;1-1amt1;3-1 showed a decreased sensitivity to the toxic ammonium analog methylammonium and a decrease in the ammonium influx of up to 70% relative to wild-type plants. These results suggest an additive contribution of AMT1;1 and AMT1;3 to the overall ammonium uptake capacity in Arabidopsis roots under nitrogen-deficiency conditions.

Arabidopsis↗

Nitrogen-deficient medium in the differential isolation of Klebsiella and Enterobacter from feces.

On a nitrogen-deficient agar medium, the tribe Klebsielleae formed large, glistening, mucoid colonies which were easily distinguished from other colony types. Of 113 Klebsielleae isolates from human feces which were characterized, Klebsiella accounted for 88% of the total; 75% were K. pneumoniae; K. ozaenae (13%) was isolated from one individual only. The remaining strains (12%) were identified as Enterobacter cloacae. Counts (for the tribe) ranged from 10(2) to 10(6), with a median of 10(4); 9 of 53 stool specimens were negative. K. pneumoniae was also isolated from 6 of 41 frozen foil-pack foods. Anaerobic studies at room temperature and 37 C revealed no appreciable differences from aerobic plates. The nitrogen-deficient medium appeared better than E M B for isolation of Klebsielleae when they were present in low numbers relative to other coliforms; slime production by Klebsielleae concomitant with minimal growth of other bacteria is involved.

Culture Media↗

The effects of nitrogen deficiency on pigments and lipids of cyanobacteria.

In contrast to what happens in higher plants and eukaryotic algae, a nitrogen deficiency during growth causes a change in pigment composition but no significant changes in whole cell lipid and fatty acid composition of the two Cyanobacteria, Pseudanabaena sp. (strain M2) and Oscillatoria splendida (strain L3). Nitrogen deficiency does not affect the cellular content in chlorophyll a, but it causes a selective loss in phycobiliproteins; carotenoid content increases with phycocyanin depletion. The major cellular lipids in both Cyanobacteria studied are monogalactosyl diacylglycerol, digalactosyl diacylglycerol, sulfoquinovosyl diacylglycerol, and phosphatidylglycerol. The fatty acid composition is particularly interesting as both these filamentous Oscillatoriaceae show important contents in alpha- and gamma-linolenic (18:3) and parinaric (18:4) acids. This seems to be very unusual in Cyanobacteria.

Journal Article↗

Performance of an aerobic/anaerobic hybrid bioreactor under the nitrogen deficient and low F/M conditions.

A bioreactor system without a biomass-liquid separation unit is evaluated for its chemical oxygen demand (COD) removal and biomass retention capabilities under the nitrogen deficient and low F/M conditions that are known to produce bulking biomass. A fully oxygenated stream recycled from an external oxygenator delivers the oxygen to an upflow bioreactor in which a biomass zone is formed and maintained in the absence of gas effervescence. COD is removed with up to 90% efficiency by means of aerobic and anaerobic bacterial activities occurring in the biomass zone. The biomass is bulking which is brought about by the extensive filamentous growth caused by the nitrogen deficient and low F/M conditions adopted. However, the biomass zone is undisturbed at superficial upflow velocities as high as 0.66 cm/min, because it has a porous, mat-like matrix that is augmented by the entanglement of filamentous bacteria with the cell clusters. A low-VSS effluent (i.e.,< 10 mg/L) is produced directly from the bioreactor.

Aerobiosis↗

Alterations in Cd-induced gene expression under nitrogen deficiency in Hordeum vulgare.

The inter-relation between nitrogen availability and cadmium toxicity was studied in roots of barley seedlings with emphasis on the analysis of expression of 10 selected genes relevant for growth in the presence of toxic Cd concentrations. The response to Cd exposure differed quantitatively or qualitatively for the 10 genes in dependence of the N supply. Transcripts of glutathione synthase, glutathione reductase, glutathione peroxidase and dehydroascorbate reductase were measured as parameters involved in antioxidant defence, metallothionein, phosphoenolpyruvate carboxylase and phytochelatin synthase (PCS) were analysed as genes related to heavy metal binding, and vacuolar ATPase subunits VHA-E and VHA-c and a NRAMP-transporter as genes being implicated in Cd transport. Reprogramming of the Cd response was most obvious for PCS and NRAMP whose transcript levels were unaltered and down-regulated, respectively, in the presence of Cd at adequate N, but strongly up-regulated upon Cd exposure under conditions of nitrogen deficiency. Different responses to Cd at varying N supply were also seen for the antioxidant genes. The results on gene expression are discussed in context with the changes in biochemical parameters, and underline the importance of evaluating the general growth conditions of a plant when discussing its specific response to a stressor such as Cd. The sequence of the nramp cDNA was filed at the EMBL/GenBank/DDBJ Databases under the accession number AJ514946.

Journal Article↗

Continuous oxidative assimilation of acetic Acid and endogenous protein synthesis applicable to treatment of nitrogen-deficient waste waters.

A previous study indicated that the oxidative assimilation capacity of a heterogeneous microbial population for acetic acid did not return to the initial value after a period of endogenous respiration in the presence of ammonium sulfate. In view of this finding it was not possible to recommend fully the use of a continuous oxidative assimilation process for the treatment of nitrogen-deficient waste waters of noncarbohydrate nature. To put the process to a severe test, a laboratory-scale pilot plant study was done. Acetate (1,000 mg/liter) was fed continuously to a completely mixed, aerated reactor vessel, from which the mixed liquor was channeled to a settling basin. The settling basin supernatant fluid was continuously discharged, and a portion of the settled biological sludge was subjected to endogenous respiration in the presence of ammonium sulfate and was recycled to the aeration vessel. Experiments were conducted at three levels of ammonia nitrogen in the endogenous phase. Approximately 90% removal of the carbon source (expressed as chemical oxygen demand, COD) was attained with a low level of supplemental nitrogen (COD/N = 70:1) and a fairly low reactor detention time (4 hr). Based upon these and previous results, it has been concluded that the process shows promise for application to the treatment of a wide variety of nitrogen-deficient industrial wastes.

Journal Article↗