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PubMed2006

Physiology and biochemistry of source-regulated protein accumulation in the wheat grain.

Wheat is unique among cereals for the baking qualities of its flour, which are dependent upon the type and concentration of its proteins. As a consequence, the grain protein concentration (GPC) is one of the main determinants of wheat international market price. More than 50-70% of the final grain N is accumulated before flowering and later remobilized to the grain, N fertilization being the common practice used to produce high GPC. However, after incremental additions of N fertilizer, GPC reaches a maximum and then remains constant, without any increase in N uptake or remobilization by the crop, thus decreasing the efficiency of N fertilizer. Although, the genetic and molecular mechanisms that regulate N uptake by the roots are being clarified quickly, the regulation and physiology of N transport from the leaves to the grain remains less clear. In this review, the possible regulatory points involved in N transport to the grain and the difficulties for increasing GPC are discussed. It has been demonstrated that protein synthesis in the grain is source-limited, and that the grain can accumulate protein limited only by the amino acids provided by the phloem. It has also been shown that there is no limitation in the amino acid/sugar ratios that can be exported to the phloem. On the other hand, NO(3)(-) uptake transporters are depressed when the plant concentration of some amino acids, such as glutamine, is high. It has also been shown that a high N supply increases cytokinins concentration, preventing leaf senescence and proteolysis. Based on this information, it is postulated that there are two main regulatory points during grain filling when plant N status is ample. On the one hand, the N uptake transporters in the roots are depressed due to the high amino acids concentration in the tissues, and N uptake is low. On the other, a high amino acids concentration keeps the cytokinins level high, repressing leaf protein degradation and decreasing amino acid export to the phloem. As a consequence, GPC cannot be increased despite the ample N supply.

Gene Expression Regulation, Plant↗

PubMed2000

Carbon dioxide uptake efficiency by outdoor microalgal cultures in tubular airlift photobioreactors.

The influence of solar irradiance and carbon dioxide molar fraction of injected CO(2)-air mixtures on the behavior of outdoor continuous cultures of the microalga Phaeodactylum tricornutum in tubular airlift photobioreactors was analyzed. Instantaneous solar irradiance, pH, dissolved oxygen, temperature, biomass concentration, and the mass flow rates of both the inlet and outlet oxygen and carbon with both the liquid and gas phases were measured. In addition, elemental analysis of the biomass and the cell-free culture medium was performed. The oxygen production rate and carbon dioxide consumption rate increased hyperbolically with the incident solar irradiance on the reactor surface. Carbon losses showed a negative correlation with the daily variation of the carbon dioxide consumption rate. The maximum CO(2) uptake efficiency was 63% of the CO(2) supplied when the CO(2) concentration in the gas supplied was 60% v/v. Carbon losses were >100% during the night, due to CO(2) production by respiration, and hyperbolically decreased to values of 10% to 20% in the midday hours. An increase in the carbon fixed in the biomass with the solar cycle was observed. A slight daily decrease of carbon content of the cell-free culture medium indicated the existence of carbon accumulation in the culture. A decrease in CO(2) molar fraction in the injected gas had a double benefit: first, the biomass productivity of the system was enhanced from 2.05 to 2.47 g L(-1) day(-1) by reduction of CO(2) inhibition and/or pH gradients; and second, the carbon losses during the daylight period were reduced by 60%. The fluid dynamics in the reactor also influenced the carbon losses: the higher the liquid flow rate the higher the carbon losses. By using a previous mass transfer model the experimental results were simulated and the usefulness of this method in the evaluation and scale-up of tubular photobioreactors was established.

Absorption↗

PubMed2002

Structure and function of shisham forests in central Himalaya, India: nutrient dynamics.

The structure and function of Shisham (Dalbergia sissoo Roxb.) forests were investigated in relation to nutrient dynamics in 5- to 15-year-old stands growing in central Himalaya. Nutrient concentrations and storage in different layers of vegetation were in the order: tree > shrub > herb. Forest soil, litter and vegetation accounted for 80.1-91.9, 1.0-1.5 and 7.0-18.4%, respectively, of the total nutrients in the system. There were considerable reductions (trees 32.8-43.1; shrubs 26.2-32.4; and herbs 18-8-22-2%) in nutrient concentrations of leaves during senescence. Nutrient uptake by the vegetation as a whole and also by the different components, with and without adjustment for internal recycling, was investigated. Annual transfer of litter nutrients to the soil from vegetation was 74.8-108.4 kg ha(-1) year(-1) N, 56.8-4 kg ha(-1) year(-1) P and 38.7-46.9 kg ha(-1) year(-1) K. Turnover rate and time for different nutrients ranged between 56 and 66 % year(-1) and 1.5 and 1.8 years, respectively. The turnover rate of litter indicates that over 50% of nutrients in litter on the forest floor are released, which ultimately enhances the productivity of the forest stand. The nutrient use efficiency in Shisham forests ranged from 136 to 143 kg ha(-1) year(-1) for N, 1,441 to 1,570 kg ha(-1) year(-1) for P and 305 to 311 kg ha(-1) year(-1) for K. Compared with natural oak forest (265 kg ha(-1) year(-1) and an exotic eucalypt plantation (18 kg ha(-1) year(-1), a higher proportion of nutrients was retranslocated in Shisham forests, largely because of higher leaf tissue nutrient concentrations. This indicates a lower nutrient use efficiency of Shisham compared with eucalypt and oak. Compartment models for nutrient dynamics have been developed to represent the distribution of nutrients pools and net annual fluxes within the system.

Australia↗

PubMed2002

Anoxic growth of phosphate-accumulating organisms (PAOs) in biological nutrient removal activated sludge systems.

In this paper, research on the growth performance of phosphate-accumulating organisms (PAOs) was conducted based on literature and experimental investigations on biological nutrient removal (BNR) activated sludge (BNRAS) systems. The research aims at presenting the occurrence of denitrifying PAOs (DPAOs), abstracting information on the kinetics and stoichiometry of PAOs under anoxic conditions and determining the conditions that stimulate the PAO growth under anoxic conditions. The research results indicate that the PAOs are capable of utilizing nitrate as electron acceptor instead of oxygen in BNRAS systems, particularly in external nitrification BNRAS (ENBNRAS) systems. However, the growth yield of PAOs under anoxic conditions should be reduced to about 70% of that under aerobic conditions, and further the stoichiometric coefficient for anoxic P uptake per PHB COD utilized should be reduced to about 80% of that under aerobic conditions as the DPAOs show a significantly lower BEPR performance and use the influent RBCOD less "efficiently" compared with aerobic PAOs (APAOs). The research results also indicate that the major factor influencing the occurrence of DPAOs and associated anoxic P uptake is the nitrate load into the anoxic reactor, i.e. the nitrate load should be large enough or exceeds the denitrification potential of ordinary heterotrophic organisms (OHOs), i.e. non-PAO organisms in the anoxic reactor to stimulate DPAOs in the system as the specific denitrification rate of OHOs (K'2 OHO) is significantly larger than that of PAOs (K'2 PAO). In terms of this competition, if the nitrate load into the main anoxic reactor is less than the denitrification potential of OHOs, then the OHOs will outcompete PAOs for using the limited nitrate, while if the nitrate load in the main anoxic reactor exceeds the denitrification potential of OHOs, then the PAOs would have opportunities to use the "excess" nitrate and so develop in the system. The other factors that influence DPAOs include the system aerobic mass fraction, sequence of reactors and frequency of sludge alternation between the aerobic and anoxic states. Although it does appear that these factors above may significantly influence the fraction of DPAOs (etaG), the quantitative relationship between these factors and etaG is not known, and the experimental observations indicate that this will be system-specific, and require calibration for each situation.

Bioreactors↗

PubMed1994

Effects of graded amounts of duodenal infusions of lysine on the mammary uptake of major milk precursors in dairy cows.

Effects of providing a limiting AA on milk secretion and mammary nutrient uptake were investigated using four lactating dairy cows given duodenal infusions of graded amounts of L-Lys-HCl (0, 9, 27, and 63 g/d). Infusions were administered over 4-d periods according to a Latin square design. Diets consisted of a 70:30 corn silage:concentrate ratio supplemented with 1.5 kg of DM/d as dehydrated alfalfa. The basal diet met requirements for energy and protein and was deficient in Lys. Cows were fitted with indwelling catheters inserted into the left carotid and left subcutaneous abdominal veins, and a transit-time flow probe was implanted around the left external pudic artery. Milk, fat, and protein yields were unaffected by infusions of Lys. Milk protein content increased to a maximum with the third treatment, and fat content showed opposite variation. Although arterial concentrations and arteriovenous differences of Lys increased to a plateau with the third treatment, the relationship between arteriovenous differences and arterial concentrations was curvilinear. Infusions of Lys tended to increase the efficiency of N utilization and the mammary extraction rates of AA to synthesize more milk proteins. Initially, Lys was extracted in a direct ratio to its milk output and then taken up in excess by the secretory cells, suggesting that Lys was no longer a limiting factor in milk protein synthesis.

Amino Acids↗

PubMed1999

Net nutrient flux in visceral tissues of lambs fed diets differing in supplemental nitrogen source.

Eight multicatheterized wethers (35.9 +/- .8 kg BW) were used in a replicated 4 x 4 Latin square design to measure N retention and net uptake and release of plasma metabolites across the portal-drained viscera (PDV), hepatic (HEP), and total splanchnic (TS) tissues in response to changes in supplemental N source. Treatments selected to provide different amounts of undegradable intake protein (UIP) were urea, soybean meal (SBM), poultry by-product meal (PBM), and bloodmeal:corn gluten meal (BMCGM; 50:50 CP basis). Diets (urea, SBM, PBM, and BMCGM) contained 12.9, 13.8, 13.6, and 13.2% CP, respectively. Periods were 10 d, with total feces and urine collected on d 7 to 10 and blood sampled on d 10. Wethers were fed at 2% of BW in 12 daily portions. Nitrogen retention was 2.2, 3.3, 4.1, and 4.4 g/d for urea, SBM, PBM, and BMCGM, respectively. Urea had less (P < .01) N retention than SBM, PBM, and BMCGM; SBM had less N retention (P < .01) than PBM and BMCGM. Arterial, portal, and hepatic plasma flows were greater (P < .09) for SBM than for PBM and BMCGM (21 vs 16, 17; 84 vs 72, 72; 105 vs 87, 88 L/h). Portal plasma flow was greater (P < .10) for urea than for SBM, PBM, and BMCGM (85 vs 84, 72, 72 L/h). Portal-drained viscera and TS alpha-amino N (AAN) fluxes were less (P < .05) for PBM than for BMCGM (20.5 vs 26.6 and 7.2 vs 15.1 mmol/h), but TS AAN flux was less (P < .05) for urea than for SBM, PBM, and BMCGM (6.9 vs 16.9, 7.2, 15.1 mmol/h). Portal-drained viscera flux and HEP removal of NH3 N were greater (P < .001) for SBM than for PBM and BMCGM (27.7 vs 19.4, 20.6; -28.1 vs -20.0, -21.4 mmol/h). Gut use was less (P = .07) and HEP and TS fluxes of urea N were greater (P < .01) for SBM than for PBM and BMCGM (-4.92 vs -8.32, -7.93; 25.87 vs 16.54, 20.00; 20.95 vs 8.22, 12.07 mmol/h). These data suggest that PBM and BMCGM improved efficiency of N use compared with urea and SBM by reducing urinary N loss.

Animal Feed↗

PubMed1999

Closed-loop control of bacterial high-cell-density fed-batch cultures: production of mcl-PHAs by Pseudomonas putida KT2442 under single-substrate and cofeeding conditions.

Pseudomonas putida KT2442 is able to accumulate medium-chain-length poly(3-hydroxyalkanoates) (mcl-PHAs) as intracellular inclusions on a variety of fatty acids and many other carbon sources. Some of these substrates, such as octanoic acid, alkenoic acids, and halogenated derivatives, are toxic when present in excess. Efficient production of mcl-PHAs on such toxic substrates therefore requires control of the carbon source concentration in the supernatant. In this study, we develop a closed-loop control system based on on-line gas chromatography to maintain continuously fed substrates at desired levels. We used the graphical programming environment LABVIEW to set up a flexible process control system that allows users to perform supervisory process control and permits remote access to the fermentation system over the Internet. Single-substrate supernatant concentration in a high-cell-density fed-batch fermentation process was controlled by a proportional (P) controller (P = 50%) acting on the substrate pump feed rate. Na-octanoate concentrations oscillated around the setpoint of 10 mM and could be maintained between 0 and 25 mM at substrate uptake rates as high as 90 mmol L(-1) h(-1). Under cofeeding conditions Na-10-undecenoate and Na-octanoate could be individually controlled at 2.5 mM and 9 mM, respectively, by applying a proportional integral (PI) controller for each substrate. The resulting copolymer contained 43.5 mol% unsaturated monomers and reflected the ratio of 10-undecenoate in the feed. It was suggested that both substrates were consumed at similar rates. These results show that this control system is suitable for avoiding substrate toxicity and supplying carbon substrates for growth and mcl-PHA accumulation.

Carbon↗

PubMed1974

Fate of donor deoxyribonucleic acid in a highly transformation-deficient strain of Haemophilus influenzae.

A transformation-deficient strain of Haemophilus influenzae (efficiency of transformation 10(4)-fold less than that of the wild type), designated TD24, was isolated by selection for sensitivity to mitomycin C. In its properties the mutant was equivalent to recA type mutants of Escherichia coli. The TD24 mutation was linked with the str-r marker (about 30%) and only weakly linked with the nov-r2.5 marker. The uptake of donor deoxyribonucleic acid (DNA) was normal in the TD24 strain, but no molecules with recombinant-type activity (molecules carrying both the donor and the resident marker) were formed. In the mutant the intracellular presynaptic fate of the donor DNA was the same as that in the transformation-proficient (wild-type) strain, and the radioactive label of the donor DNA associated covalently with the recipient chromosome in about the same quantity as in the wild type. However, many fewer donor atoms were associated with segments of the mutant's recipient chromosome as compared with segments of the wild-type chromosome. In the mutant the association was accompanied by complete loss of donor marker activity. The lack of donor marker activity of the donor-recipient complex of DNA isolated from the mutant was not due to lack of uptake of the complex by the second recipient and its inability to associate with the second recipient's chromosome. Because the number of donor-atom-carrying resident molecules was higher than could be accounted for by the lengths of presynaptic donor molecules, we favor the idea that the association of donor DNA atoms with the mutant chromosome results from local DNA synthesis rather than from dispersive integration of donor DNA by recombination.

Centrifugation, Density Gradient↗

PubMed1999

The phosphotransferase system (PTS) of Streptomyces coelicolor identification and biochemical analysis of a histidine phosphocarrier protein HPr encoded by the gene ptsH.

HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS) controls sugar uptake and carbon utilization in low-GC Gram-positive bacteria and in Gram-negative bacteria. We have purified HPr from Streptomyces coelicolor cell extracts. The N-terminal sequence matched the product of an S. coelicolor orf, designated ptsH, sequenced as part of the S. coelicolor genome sequencing project. The ptsH gene appears to form a monocistronic operon. Determination of the evolutionary relationship revealed that S. coelicolor HPr is equally distant to all known HPr and HPr-like proteins. The presumptive phosphorylation site around histidine 15 is perfectly conserved while a second possible phosphorylation site at serine 47 is not well-conserved. HPr was overproduced in Escherichia coli in its native form and as a histidine-tagged fusion protein. Histidine-tagged HPr was purified to homogeneity. HPr was phosphorylated by its own enzyme I (EI) and heterologously phosphorylated by EI of Bacillus subtilis and Staphylococcus aureus, respectively. This phosphoenolpyruvate-dependent phosphorylation was absent in an HPr mutant in which histidine 15 was replaced by alanine. Reconstitution of the fructose-specific PTS demonstrated that HPr could efficiently phosphorylate enzyme IIFructose. HPr-P could also phosphorylate enzyme IIGlucose of B. subtilis, enzyme IILactose of S. aureus, and IIAMannitol of E. coli. ATP-dependent phosphorylation was detected with HPr kinase/phosphatase of B. subtilis. These results present the first identification of a gene of the PTS complement of S. coelicolor, providing the basis to elucidate the role(s) of HPr and the PTS in this class of bacteria.

Adenosine Triphosphate↗

PubMed2004

Interaction of flooding with carbon metabolism of forest trees.

Waterlogging and flooding cause oxygen deprivation in the root system of trees. Since oxygen is essentially for mitochondrial respiration, this process cannot be maintained under anoxic conditions and must be replaced by other pathways. For the roots it is therefore a matter of survival to switch from respiration to alcoholic fermentation. Due to the low efficiency of this process to yield energy equivalents (ATP), energy and carbon metabolism of trees are usually strongly affected by oxygen deprivation, even if a rapid switch from respiration to fermentation is achieved. The roots can compensate for the low energy yield of fermentation either (1) by decreasing the demand for energy by a reduction of energy-dependent processes such as root growth and/or nutrient uptake, or (2) by consuming more carbohydrates per unit time in order to generate sufficient energy equivalents. In the leaves of trees, flooding and waterlogging cause a decline in the rates of photosynthesis and transpiration, as well as in stomatal conductance. It is assumed that, due to reduced phloem transport, soluble sugars and starch accumulate in the leaves of flooded trees, thereby negatively affecting the sugar supply of the roots. Thus, root growth and survival is negatively affected by both changes in root internal carbon metabolism and impaired carbon allocation to the roots by phloem transport. In addition, accumulation of toxic products of fermentation in the roots, such as acetaldehyde, can further impair root metabolism. A main feature of tolerance against flooding and waterlogging of trees seems to be the steady supply of carbohydrates to the roots in order to maintain alcoholic fermentation; in addition, roots of tolerant trees seem to avoid accumulation of fermentation-derived ethanol and acetaldehyde. From studies with flooding tolerant and non-tolerant tree species, it is hypothesized that (1) the transport of ethanol produced in the roots under hypoxic conditions into the leaves via the transpiration stream, (2) its conversion into acetyl-CoA in the leaves, and (3) its use in the plant's general metabolism, are mechanisms of flooding tolerance of trees.

Adaptation, Physiological↗