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Leaf canopy as a dynamic system: ecophysiology and optimality in leaf turnover.

BACKGROUND AND AIMS: In a leaf canopy, there is a turnover of leaves; i.e. they are produced, senesce and fall. These processes determine the amount of leaf area in the canopy, which in turn determines canopy photosynthesis. The turnover rate of leaves is affected by environmental factors and is different among species. This mini-review discusses factors responsible for leaf dynamics in plant canopies, focusing on the role of nitrogen. SCOPE: Leaf production is supported by canopy photosynthesis that is determined by distribution of light and leaf nitrogen. Leaf nitrogen determines photosynthetic capacity. Nitrogen taken up from roots is allocated to new leaves. When leaves age or their light availability is lowered, part of the leaf nitrogen is resorbed. Resorbed nitrogen is re-utilized in new organs and the rest is lost with dead leaves. The sink-source balance is important in the regulation of leaf senescence. Several models have been proposed to predict response to environmental changes. A mathematical model that incorporated nitrogen use for photosynthesis explained well the variations in leaf lifespan within and between species. CONCLUSION: When leaf turnover is at a steady state, the ratio of biomass production to nitrogen uptake is equal to the ratio of litter fall to nitrogen loss, which is an inverse of the nitrogen concentration in dead leaves. Thus nitrogen concentration in dead leaves (nitrogen resorption proficiency) and nitrogen availability in the soil determine the rate of photosynthesis in the canopy. Dynamics of leaves are regulated so as to maximize carbon gain and resource-use efficiency of the plant.

Adaptation, Physiological↗

Influence of microplastics on microalgal performance during wastewater polishing.

Microplastics (MPs) are emerging contaminants that are increasingly accumulating in aquatic ecosystems due to excessive anthropogenic activity and insufficient mitigation strategies, posing serious environmental and public health risks. Their impact on wastewater (WW) treatment processes remains poorly understood. This study evaluated the effects of five MPs commonly found in WW - polypropylene, polystyrene, polyamide, low-density polyethylene, and high-density polyethylene - on the physiology and bioremediation performance of the microalga Chlorella vulgaris in synthetic WW (SWW). Metabolic responses were assessed via esterase activity and intracellular reactive oxygen species (ROS), while nitrogen (N), phosphorus (P), and glucose removal were monitored to evaluate bioremediation efficiency. MPs inhibited esterase activity and elevated ROS levels, indicating oxidative stress. Nevertheless, C. vulgaris maintained a high bioremediation capacity (> 75 % N, > 60 % P, and > 70 % for glucose). Environmental conditions modulated microalga response to MPs exposure. Under N-limited conditions, C. vulgaris exhibited enhanced nutrient uptake and biomass production, but a 12 h/12 h light/dark photoperiod reduced N removal but stimulated glucose consumption via heterotrophic metabolism. In contrast, C-limited conditions exacerbated oxidative stress and compromised nutrient removal, resulting in residual concentrations exceeding legal limits. These findings highlight that environmental factors can either mitigate or exacerbate the physiological stress induced by MPs, ultimately affecting WW polishing. This work provides a comprehensive insight into the cellular and metabolic effects of MPs on microalgae and supports C. vulgaris as a resilient and sustainable approach for nutrient and carbon removal in MP-contaminated WW systems.

Microalgae↗

[Nitrogen and phosphorus removal from septage filtrate in a ryegrass hydroponic system].

With hydroponic method, this paper studied the ryegrass Lolium multiflorum growth and its N and P removal under the application of septage filtrate. The results showed that a medium application rate of septage filtrate gave the highest yield of L. multiflorum. The N and P removal in the ryegrass hydroponic system was increased with increasing application rate of septage filtrate, but the water quality was declined when super high volume of septage filtrate was added. The N and P uptake by L. multiflorum had no significant difference under medium, high, and super high application rate of septage filtrate, with the maximum uptake of N and P being 13.6 g x m(-2) and 3.17 g x m(-2) respectively. The removed proportion of N and P through ryegrass uptake declined w ith increasing septage filtrate application rate. Under the same total load, increasing the application frequency of septage filtrate could improve ryegrass yield, water quality, and nutrients removal. To select an appropriate application rate and frequency of septage filtrate was the key in improving the nutrients removal efficiency of ryegrass hydroponic system.

Biodegradation, Environmental↗

Nocturnal uptake and assimilation of nitrogen dioxide by C3 and CAM plants.

In order to investigate nocturnal uptake and assimilation of NO2 by C3 and crassulacean acid metabolism (CAM) plants, they were fumigated with 4 microl l(-1) 15N-labeled nitrogen dioxide (NO2) for 8 h. The amount of NO2 and assimilation of NO2 by plants were determined by mass spectrometry and Kjeldahl-nitrogen based mass spectrometry, respectively. C3 plants such as kenaf (Hibiscus cannabinus), tobacco (Nicotiana tabacum) and ground cherry (Physalis alkekengi) showed a high uptake and assimilation during daytime as high as 1100 to 2700 ng N mg(-1) dry weight. While tobacco and ground cherry strongly reduced uptake and assimilation of NO2 during nighttime, kenaf kept high nocturnal uptake and assimilation of NO2 as high as about 1500 ng N mg(-1) dry weight. Stomatal conductance measurements indicated that there were no significant differences to account for the differences in the uptake of NO2 by tobacco and kenaf during nighttime. CAM plants such as Sedum sp., Kalanchoe blossfeldiana (kalanchoe) and Aloe arborescens exhibited nocturnal uptake and assimilation of NO2. However, the values of uptake and assimilation of NO2 both during daytime and nighttime was very low (at most about 500 ng N mg(-1) dry weight) as compared with those of above mentioned C3 plants. The present findings indicate that kenaf is an efficient phytoremediator of NO2 both during daytime and nighttime.

Aloe↗

Components of a ubiquitin ligase complex specify polyubiquitination and intracellular trafficking of the general amino acid permease.

Gap1p, the general amino acid permease of Saccharomyces cerevisiae, is regulated by intracellular sorting decisions that occur in either Golgi or endosomal compartments. Depending on nitrogen source, Gap1p is transported to the plasma membrane, where it functions for amino acid uptake, or to the vacuole, where it is degraded. We found that overexpression of Bul1p or Bul2p, two nonessential components of the Rsp5p E3-ubiquitin ligase complex, causes Gap1p to be sorted to the vacuole regardless of nitrogen source. The double mutant bul1Delta bul2Delta has the inverse phenotype, causing Gap1p to be delivered to the plasma membrane more efficiently than in wild-type cells. In addition, bul1Delta bul2Delta can reverse the effect of lst4Delta, a mutation that normally prevents Gap1p from reaching the plasma membrane. Evaluation of Gap1p ubiquitination revealed a prominent polyubiquitinated species that was greatly diminished in a bul1Delta bul2Delta mutant. Both a rsp5-1 mutant and a COOH-terminal truncation of Gap1p behave as bul1Delta bul2Delta, causing constitutive delivery of Gap1p to the plasma membrane and decreasing Gap1p polyubiquitination. These results indicate that Bul1p and Bul2p, together with Rsp5p, generate a polyubiquitin signal on Gap1p that specifies its intracellular targeting to the vacuole.

Adaptor Proteins, Signal Transducing↗

Auxotrophic mutant of the cyanobacterium Nostoc muscorum showing absolute requirement of Cs+ or Rb+ for diazotrophy and autotrophy.

Caesium-resistant (Cs(+)-R) mutant clones of the cyanobacterium Nostoc muscorum were characterized for diazotrophic growth in a medium devoid of Cs(+) or Rb(+) or both. Cs(+)-R phenotype suffered severe genetic damage of a pleiotropic nature affecting diazotrophic growth, chlorophyll a content, nitrogenase activity and photosynthetic O(2) evolution. Mutation leading to development of Cs(+)-R phenotype could be overcome by availability of Cs(+)/Rb(+). Parent and mutant strains were similar with respect to their Cs(+)/Rb(+) uptake. Available data suggests operation of an efficient coupling of the two incompatible reactions viz. oxygenic photosynthesis and oxygen sensitive N(2) fixation in this cyanobacterium.

Cesium↗

Novel effects of a transposon insertion in the Vibrio fischeri glnD gene: defects in iron uptake and symbiotic persistence in addition to nitrogen utilization.

Vibrio fischeri is the sole species colonizing the light-emitting organ of the Hawaiian squid, Euprymna scolopes. Upon entering the nascent light organ of a newly hatched juvenile squid, the bacteria undergo morphological and physiological changes that include the loss of flagellation and the induction of bioluminescence. These and other events reveal a pattern of genetic regulation that is a response to the colonization of host tissue. In this study, we isolated and characterized a glnD:mTn5Cm mutant of V. fischeri. In addition to the predicted defects in the efficiency of nitrogen utilization, this glnD mutant had an unexpected reduction in the ability to produce siderophore and grow under iron-limiting conditions. Although the glnD mutant could colonize juvenile squid normally over the first 24 h, it was subsequently unable to persist in the light organ to the usual extent. This persistence phenotype was more severe if the mutant was pregrown under iron-limiting conditions before inoculation, but could be ameliorated by the presence of excess iron. These results indicate that the ability to respond to iron limitation may be an important requirement in the developing symbiosis. Supplying the glnD gene in trans restored normal efficiency of nitrogen use, iron sequestration and colonization phenotypes to the glnD:mTn5Cm mutant; thus, there appears to be a genetic and/or metabolic linkage between nitrogen sensing, siderophore synthesis and symbiosis competence in V. fischeri that involves the glnD gene.

Animals↗

California black oak response to nitrogen amendment at a high O3, nitrogen-saturated site.

In a nitrogen (N) saturated forest downwind from Los Angeles, California, the cumulative response to long-term background-N and N-amendment on black oak (Quercus kelloggii) was described in a below-average and average precipitation year. Monthly measurements of leaf and branch growth, gas exchange, and canopy health attributes were conducted. The effects of both pollutant exposure and drought stress were complex due to whole tree and leaf level responses, and shade versus full sun leaf responses. N-amended trees had lower late summer carbon (C) gain and greater foliar chlorosis in the drought year. Leaf water use efficiency was lower in N-amended trees in midsummer of the average precipitation year, and there was evidence of poor stomatal control in full sun. In shade, N-amendment enhanced stomatal control. Small differences in instantaneous C uptake in full sun, lower foliar respiration, and greater C gain in low light contributed to the greater aboveground growth observed.

Acclimatization↗

Postprandial intestinal and whole body nitrogen kinetics and distribution in piglets fed a single meal.

Our aim was to characterize the postprandial total and dietary N fluxes in the portal drained viscera (PDV) and whole body after administration of a single meal in young pigs. Seven 4-wk-old piglets, implanted with a portal flow probe and portal, arterial and venous catheters, received a primed constant [(18)O]urea intravenous infusion and were studied for 8 h after a bolus mixed meal ingestion (46 mmol N/kg body wt) intrinsically labeled with (15)N to trace dietary N fluxes. The real cecal digestibility of the formula was 94.3% (SD 1.8). PDV output of dietary N was found principally in the pool of circulating protein (51% of the measured dietary N PDV output), in the free alpha-amino N pool (44%), and to a lesser extent in ammonia (5%). Dietary N release in alpha-amino N and ammonia mainly occurred during the first 3 h. Total and exogenous postprandial urea productions were 5.8 and 2.0 mmol N/kg body wt, respectively. At the end of the postprandial period, losses of dietary N amounted to 10.3% of the dose: 5.7% through ileal losses and 4.6% by deamination and transfer to urea. Net postprandial retention of dietary N was 90.4% (SD 1.3), of which 20% was found in splanchnic zone (small intestine 10%, liver 5%, and plasma protein 3%) and 42% in peripheral zone (muscle 31%, skin 6%). In conclusion, our results show a high efficiency of dietary N utilization for muscular uptake and anabolic utilization. However, the results obtained point out the necessity to further explore the form of dietary N released into the portal blood.

Adaptation, Physiological↗

Substrate specificity and expression profile of amino acid transporters (AAPs) in Arabidopsis.

Three amino acid transporter genes (AAP3-5) were isolated from Arabidopsis by complementation of a yeast mutant defective in histidine uptake. Transport is driven against a concentration gradient and sensitive to protonophores. Analysis of the substrate specificity demonstrates that the carriers have a broad substrate specificity covering the major transport forms of reduced nitrogen, i.e. glutamine and glutamate. The transporters have similar affinities for glutamate, glutamine, and alanine but differ with respect to valine, phenylalanine, histidine, arginine, and lysine. AAP3 and AAP5 efficiently transport arginine and lysine and are involved in basic amino acid transport. The predicted polypeptides of 53 kDa are highly hydrophobic with 12 putative membrane-spanning regions and show significant homologies to Arabidopsis amino acid transporters AAP1 and AAP2. Each of the genes has a different organ-specific expression in the plant. AAP3 is exclusively expressed in roots and AAP4 mainly in source leaves, stems, and flowers, whereas AAP5 is found in all tissues. The specific distribution in the plant and the different substrate specificities of AAP transporters may indicate that tissues differ both qualitatively and quantitatively regarding import or export of amino acids.

Amino Acid Sequence↗

Influence of porosity and composition of porous carriers on the uptake of nutrients.

The current paper assesses the potential of industrial solid wastes utilization such as blast furnace slag (BFS) and zeolite synthesized from fly ash (ZFA), which are effective as well as economically attractive for the uptake of phosphate and ammonium from polluted seawater. The solidification of BFS and ZFA has been developed in different proportions of BFS/ZFA (30/70, 50/50 and 70/30 (w/w)) with different porosities (25%, 40% and 52%, respectively) to cylindrical porous carriers using a Hydrothermal Hot-Pressing (HHP) method. The concentrations of heavy metals in ZFA and BFS were too low to affect the aquatic environment. The main finding is that the high rate of BFS (70%) in porous carriers enhanced phosphate uptake explained by the higher percentage of calcium (35.7%) in porous carriers and high pH conditions. The efficient ammonium uptake was observed with high rate of ZFA (70%) in porous carriers. Results found through this experimental work imply that porous carriers with BFS/ZFA proportion of 70/30 are suitable for potential practical application in the aquatic environment due to their efficient uptake of phosphate and ammonium. The choice was made upon their porosity (40%) and their compressive strength (56 kgf/cm2) which are relatively higher than those with BFS/ZFA proportion of 30/70 and 50/50.

Ammonia↗

Aspartate transport by the Dct system in Rhizobium leguminosarum negatively affects nitrogen-regulated operons.

Amino acid uptake by the general amino acid permease (Aap) of Rhizobium leguminosarum strain 3841 was severely reduced by the presence of aspartate in the growth medium when glucose was the carbon source. The reduction in transport by the Aap appeared to be caused by inhibition of uptake and not by transcriptional repression. However, as measured with lacZ fusions, the Ntr-regulated gene glnII was repressed by aspartate. The negative regulatory effect on both the Aap and glnII was prevented by mutation of any component of the dicarboxylate transport (Dct) system or by the inclusion of a C4- dicarboxylate in the growth medium, including the non-metabolizable analogue 2-methylsuccinate. As measured by total uptake and with a dctA-lacZ fusion, aspartate was an efficient inducer of the Dct system, but slightly less so than succinate alone or succinate and aspartate together. Thus, aspartate does not cause overexpression of DctA leading to improper regulation of other operons. Transport measurements revealed that the Dct system has an apparent Km for succinate of 5 microM and an apparent Ki for aspartate inhibition of succinate uptake of 5 mM. These data imply that the Dct-mediated accumulation of aspartate causes an unregulated build-up of aspartate or a metabolic product of it in the cell. This accumulation of aspartate is prevented either by mutation of the dct system or by the presence of a higher affinity substrate that will reduce access of aspartate to the carrier protein. Elevation or disruption of the intracellular aspartate pool is predicted to disrupt N-regulated operons and nitrogen fixation.

Amino Acid Transport Systems↗

Technetium-99m labelled integrated tropane-BAT as a potential dopamine transporter tracer.

To reduce the molecular weight of 99mTc-labelled tropanes with the aim to enhance the passage over the blood-brain barrier, a so-called integrated tropane-BAT construct was developed. For this purpose a mercaptoethyl substituent was attached to the amine nitrogen atom of a nortropane precursor and the methyl carboxylate in 2beta-position was converted to a 2-mercaptoethylaminomethylene substituent. This integrated tropane-BAT construct could be labelled efficiently (85-90%) with technetium-99m. Results of LC-MS analysis of the tracer agent support the assumed structure. Biodistribution studies in normal rats (n=3) showed a slightly higher brain uptake for the new tracer agents as compared to 99mTc-TRODAT-1. These results indicate that further biological evaluation of the integrated 99mTc-tropane-BAT is warranted.

Animals↗

Improvement of CHO cell culture medium formulation: simultaneous substitution of glucose and glutamine.

The formulation of the culture medium for a Chinese hamster ovary (CHO) cell line has been investigated in terms of the simultaneous replacement of glucose and glutamine, the most commonly employed carbon and nitrogen sources, pursuing the objective of achieving a more efficient use of these compounds, simultaneously avoiding the accumulation of lactate and ammonium in the medium. The key factor in this process is the selection of compounds that are slowly metabolized. Among the different compounds studied, galactose and glutamate provide the best results, allowing support of cell growth with an optimal balance between nutrient uptake and cell requirements and the generation of minimal quantities of lactate and ammonium. The attained results also highlight the capacity of the cells to redistribute their metabolism as a response to the changes in medium composition.

Animals↗

Heterotrophic nutrition of the marine pennate diatom Navicula pavillardi Hustedt.

Navicula pavillardi Hustedt, a marine, littoral, pennate diatom, can grow in the dark on glutamate or on the complex organic supplements tryptone or yeast extract. Growth on glutamate in the dark took place without an initial lag phase, whereas growth on tryptone began only after a 2-day lag phase that could be abolished by the simultaneous presence of glucose. Lactate inhibited growth in the dark on glutamate, but not photoautotrophic growth. Relatively low concentrations of glutamine inhibited photoautotrophic growth. The observed doubling time for heterotrophic growth on glutamate or tryptone was about 70 h, compared with a doubling time of 24 h under optimal photoautotrophic conditions. Glucose did not decrease the doubling time in the dark on tryptone. The assimilation efficiency for glutamate was 41%. The estimated necessary uptake rate for glutamate to account for the observed heterotrophic doubling time on glutamate was close to those measured with isotope techniques. The kinetic parameters for glutamate uptake, which followed Michelis-Menten kinetics, were Ks = 0.018 mM, and Vmax = 7.0 X 10(-10) mumol per cell per minute. Although several amino acids served as sole nitrogen sources for photoautotrophic growth and were demonstrated by the use of isotope techniques to enter the cells, they could not be used as substrates for growth in the dark. Glucose was not taken up to a significant extent except by cells grown in the presence of tryptone. Lactate was taken up only by dark-grown cells. Results of preliminary studies on the metabolic fate of several uniformly labeled amino acids are presented.

Acetates↗

Generation of new hydrogen-recycling Rhizobiaceae strains by introduction of a novel hup minitransposon.

Hydrogen evolution by nitrogenase is a source of inefficiency for the nitrogen fixation process by the Rhizobium-legume symbiosis. To develop a strategy to generate rhizobial strains with H(2)-recycling ability, we have constructed a Tn5 derivative minitransposon (TnHB100) that contains the ca. 18-kb H(2) uptake (hup) gene cluster from Rhizobium leguminosarum bv. viciae UPM791. Bacteroids from TnHB100-containing strains of R. leguminosarum bv. viciae PRE, Bradyrhizobium japonicum, R. etli, and Mesorhizobium loti expressed high levels of hydrogenase activity that resulted in full recycling of the hydrogen evolved by nitrogenase in nodules. Efficient processing of the hydrogenase large subunit (HupL) in these strains was shown by immunoblot analysis of bacteroid extracts. In contrast, Sinorhizobium meliloti, M. ciceri, and R. leguminosarum bv. viciae UML2 strains showed poor expression of the hup system that resulted in H(2)-evolving nodules. For the latter group of strains, no immunoreactive material was detected in bacteroid extracts using anti-HupL antiserum, suggesting a low level of transcription of hup genes or HupL instability. A general procedure for the characterization of the minitransposon insertion site and removal of antibiotic resistance gene included in TnHB100 has been developed and used to generate engineered strains suitable for field release.

Bacterial Proteins↗

Nitrogen use scenario in India.

Nitrogen is one of the major plant nutrients without which the agricultural production is not possible. Nitrogen use in Indian agriculture was nearly 55000 tons in 1950-1951 that increased to 11.31 million tons in 2001-2002. The total food production of the country has also experienced the similar increase from 50.83 to 222 million tons in the respective years. Interestingly the N fertilizer consumption of India remained almost constant during the last six years indicating the possibility of reducing N consumption. The highest N consumption is in North zone owing to the introduction of rice-wheat cropping system followed by West, South and East. The N use efficiency has been reported to be varying between 30% to 50% depending on the crops and the management. But in most of the cases, N use efficiency has been calculated based on the total N removed by the crops (above ground part only) ignoring the N content left in the roots. It has been observed in controlled experiments that the total N uptake by roots varied from 18% to 44% of the total N removed by the above ground parts, i.e. grain and straw. If the root N is also accounted, the N use efficiency will be higher than reported. The management of other organic sources has to be improved so as to increase the fertilizer use efficiency as well as to check the direct release of N in the atmosphere. In this review all these issues will be dealt.

Agriculture↗

The role of DPB at anoxic stage in a novel package type BNR process with batch settler.

In this research, package type BNR process was developed to remove nutrients as well as suspended solids from domestic sewage. The effect of HRT reduction of settler on the removal of pollutants as well as the role of DPBs at anoxic reactor were investigated. The proposed package system was composed of sludge denitrification tank, anaerobic, anoxic, oxic, and batch settler. This system could remove nitrogen and phosphorus effectively at low COD/N ratio and also remove SS more effectively than other ordinary BNR system having the conventional settlers. The removal efficiency of total nitrogen (T-N) in optimal condition was about 75.1% under the TCOD/TKN ratio as low as 5.7. The average concentration of the effluent TCOD, ortho-P, NH4+-N, NO3- -N and SS in the package type BNR system of 2 h settler retention time were 15.6. 1.38, 1.4, 10.3 and 3.9 mg/L, respectively. In anoxic state, denitrifying dephosphatation, that is uptaking phosphate by using nitrate-nitrogen as an electron acceptor was observed. The removed NO3- -N concentration by denitrifying dephosphatation was 1.62 mg NO3- -N/mg PO4(3-)-P.

Bacteria, Anaerobic↗