Modeling linear and variable growth in phosphate limited suspension cultures of Opium poppy

In examining the growth kinetics of cell suspensions of opium poppy (Papaver somniferum), the increase in biomass with time was observed to be linear over the entire batch growth period of up to 20 days. Although batch growth profiles were reproducible utilizing the same inoculum, growth rates varied tremendously when experiments were inoculated with cells from different flasks. Both of these phenomena are difficult to explain with conventional batch growth models. In a series of a experiments, phosphate was determined to be the growth-rate-limiting substrate. By expressing growth rate in terms of the intracellular reserves of phosphorus, a growth model which expresses kinetics in terms of the intracellular phosphorus contents of the cells is shown to predict both linear growth character and inoculum dependent variability in growth. The stationary phase phosphate content of seven plant suspension cultures of different plant species was found to be comparable to phosphorus levels of phosphate-starved poppy cells, which suggests that phosphate limitation may be common for plant tissue culture. The applicability of this model to other biological systems which display similar batch growth patterns when subjected to inorganic nutrient deprivation is discussed.     

Root developmental adaptation to phosphate starvation: better safe than sorry

Phosphorus is a crucial component of major organic molecules such as nucleic acids, ATP and membrane phospholipids. It is present in soils in the form of inorganic phosphate (Pi), which has low availability and poor mobility. To cope with Pi limitations, plants have evolved complex adaptive responses that include morphological and physiological modifications. This review describes how the model plant Arabidopsis thaliana adapts its root system architecture to phosphate deficiency through inhibition of primary root growth, increase in lateral root formation and growth and production of root hairs, which all promote topsoil foraging. A better understanding of plant adaptation to low phosphate will open the way to increased phosphorus use efficiency by crops. Such an improvement is needed in order to adjust how we manage limited phosphorus stocks and to reduce the disastrous environmental effects of phosphate fertilizers overuse.     

A study of phosphate limitation in Lake Maarsseveen: phosphate uptake kinetics versus bioassays

In order to assess possible phosphate limitation for the phytoplankton community of Lake Maarsseveen, two techniques (phosphate uptake experiments and bioassays) were employed simultaneously in February–March 1982. In that period the ambient phosphate concentration of the lake water was less than 0.03 µM P and the diatom Asterionella formosa constituted more than 90% of the phytoplankton population. The phosphate uptake experiments showed relatively high uptake capacities and low cell phosphorus contents for the natural phytoplankton community. This suggested phosphate limitation throughout the test period. The growth stimulation of the phytoplankton after enrichment with phosphate, however, only revealed phosphate limitation from the beginning of March and bioassays may therefore be regarded as a less sensitive method.     

Regulation of phosphate uptake kinetics inOscillatoria agardhii

In order to study phosphate uptake kinetics the cyanobacteriumOscillatoria agardhii was grown in continuous culture under a phosphorus limitation. The affinity of the uptake system reflected in the initial slope of the uptake rate versus external substrate concentration curve (dV/ds) was found to be unaffected by the growth wate.The maximum phosphate uptake rate (V _m ) decreased as the growth rate was increased. Attempts were made to relate the decrease ofV _m to the increase in phosphorus content of the cells that occurred a higher growth rates. Accumulation of phosphate during pulse experiments indeed resulted in a decrease ofV _m . However feedback regulation ofV _m by accumulated phosphorus was found to occur only to a small extent in steady state growing cells. The main part of the regulation of the activity of the phosphate uptake system seemingly is determined by a long term process that is, at least longer than 2 h. The presence of short term feedback inhibition by accumulated phosphorus on the activity of the uptake system provides an explanation of the phenomenon thatOscillatoria agardhii is not able to grow at near _max growth rates under a phosphorus limitation.     

The influence of radioactive phosphate level on the absorption of phosphate by plants and on the determination of labile soil phosphate

Summary Previous work has suggested that the presence of P³² in fertilizers inhibits the uptake of the applied phosphate from the soil by plants, and also that if the applied phosphate is not incorporated uniformly in the soil there will be preferential uptake from regions of low specific activity. This made it desirable to determine the effect of P³²-level on phosphate uptake and the determination ofL-values in pot experiments in which the labelled phosphate source is added as discrete particles of the phosphate form of an anion-exchange resin.Increasing the level of P³² from 0.05 to 1.25 mo per gram of phosphorus in the added phosphate did not have a significant effect on the fresh weight, dry weight or total phosphorus uptake of the ryegrass crop. The measuredL-value showed a significant increase, about 15 per cent for a five-fold increase in P³² level, on each of the four soil types used, as would be expected if P³² depressed the uptake of labelled fertilizer phosphate.Although a significant effect of P³² was observed this does not invalidate a comparison of soils with respect toL-value.     

Modelling the contribution of arbuscular mycorrhizal fungi to plant phosphate uptake

In this paper, we investigate the role of arbuscular mycorrhizal fungi in plant phosphorus nutrition. We develop a mathematical model which quantitatively assesses the contribution of external fungal hyphae to plant phosphate uptake.We derive an equation for solute uptake by a growing fungal mycelium which we couple with a model for root uptake. We analyse the model using nondimensionalization and numerical simulations.Simulations predict that removal of phosphate from soil is dominated by hyphal uptake as opposed to root uptake. Model analysis shows that the depletion zones around hyphae overlap within 8 h and that the transfer between fungus and root is a critical step for the behaviour of phosphorus within the mycelial phase. We also show that the volume fraction of mycelium is negligibly small in comparison to other soil phases.This is the first model to quantify the contribution of mycorrhizal fungi to plant phosphate uptake. A full data set for model parametrization and validation is not currently available. Therefore, more complete sets of experimental measurements are necessary to make this model more applicable.     

Cell wall teichoic acid as a reserve phosphate source in Bacillus subtilis.

Although exponential growth of Bacillus subtilis 168 in a phosphate-limited medium halted with the exhaustion of inorganic phosphate, the bacteria continued to grow at a slower rate for a further 3 to 4 h at 37 degrees C. This postexponential growth in the absence of an exogenous phosphate supply was accompanied by a loss of teichoic acid from the cell walls of the bacteria. Quantitative analysis of walls and culture fluids showed that the phosphate loss from the walls could not be accounted for by an increase in phosphate-containing compounds in the medium, which implied that the cells were using their own wall teichoic acids to supply phosphate necessary for growth. Addition of exogenous teichoic acid to phosphate-starved cultures resulted in stimulation of growth and in the simultaneous disappearance of teichoic acid phosphate from the medium. It is proposed that teichoic acids, which can contain more than 30% of the total phosphorus of exponential-phase cells, can be used as a reserve phosphate source when the bacteria are starved for inorganic phosphate.     

The regulator gene phoB mediates phosphate stress-controlled synthesis of the membrane lipid diacylglyceryl-N,N,N-trimethylhomoserine in Rhizobium (Sinorhizobium) meliloti

Bacteria react to phosphate starvation by activating genes involved in the transport and assimilation of phosphate as well as other phosphorous compounds. Some soil bacteria have evolved an additional mechanism for saving phosphorous. Under phosphate-limiting conditions, they replace their membrane phospholipids by lipids not containing phosphorus. Here, we show that the membrane lipid pattern of the free-living microsymbiotic bacterium Rhizobium (Sinorhizobium) meliloti is altered at low phosphate concentrations. When phosphate is growth limiting, an increase in sulpholipids, ornithine lipids and the de novo synthesis of diacylglyceryl trimethylhomoserine (DGTS) lipids is observed. Rhizobium meliloti phoCDET mutants, deficient in phosphate uptake, synthesize DGTS constitutively at low or high medium phosphate concentrations, suggesting that reduced transport of phosphorus sources to the cytoplasm causes induction of DGTS biosynthesis. Rhizobium meliloti phoU or phoB mutants are unable to form DGTS at low or high phosphate concentrations. However, the functional complementation of phoU or phoB mutants with the phoB gene demonstrates that, of the two genes, only intact phoB is required for the biosynthesis of the membrane lipid DGTS.     

The effects of organic and inorganic pollutants on intracellular phosphate compounds in blue mussels (Mytilus edulis).

1. The effects of sublethal concentrations of organic and inorganic pollutants on intracellular energy-rich phosphates in blue mussels, Mytilus edulis, were investigated by in vivo 31P-NMR. 2. Formaldehyde (30 and 10 mg/l), phenol, pyridine, mercury and cadmium gave marked reductions in phosphoarginine and, in some cases, the ATP amounts. The reduction in high-energy phosphate was accompanied by an increase in inorganic phosphate in all groups. 3. A "phosphorus index", the product of the ratios between phosphoarginine and inorganic phosphate, and ATP and inorganic phosphate, is suggested, which might serve as an early warning ("alarm") parameter in environmental monitoring. 4. Diversity in the responses to different pollutants make phosphorus compounds in M. edulis also an interesting element in a finger print parameter system designed to distinguish between pollutants in the marine environment.     

Levels of phosphate esters in spirodela

The duckweed Spirodela oligorrhiza was grown in sterile nutrient solutions that contained 1 mm phosphate-(32)P at various specific activities. In solutions with activities higher than 2 muc per mumole per ml, plant growth was inhibited after a time, and the physical appearance of the plants was affected. The critical level of radiation, at which growth was first affected, corresponded to 5 kilorads.Plants were grown for 9 days (5 generations) in a culture solution containing phosphate at 0.5 muc per mumole per ml (radiation load approx 0.5 kilorads) so that all phosphorus-containing materials in the tissue became uniformly labeled. The various radioactive compounds were extracted, chromatographed, identified, and their radioactivity was measured. From this radioactivity plus the specific activity of the supplied phosphate, the amount of each compound was calculated. The data constitute a complete balance-sheet for phosphorus in a plant tissue. The identity of 98% of the phosphorus in the tissue was determined. Inorganic phosphate (32,700 mmumoles/g fr wt) was the predominant phosphorus-containing compound; RNA (5100 mmumoles P/g fr wt) was the main organic phosphate; phosphatidyl choline (1600 mmumoles/g fr wt) was the main phospholipid, and glucose-6-phosphate (500 mmumoles/g fr wt) the main acid-soluble phosphate ester. Amounts of other phosphorus compounds are given.     

Response of Thiobacillus ferrooxidans to phosphate limitation

Abstract: Phosphate ion is an essential nutrient for all cells. Consequently, starvation for this component may constitute a stressing condition which affects the bioleaching capacity of the biomining microorganisms. Therefore, we have studied the manner in which the chemolithotroph Thiobacillus ferrooxidans responds to phosphate limitation. Under these circumstances the bacteria reduced its growth rate, capacity to oxidize ferrous iron and to fix CO,. Concomitant with these changes, the cells showed an increased synthesis of several proteins, some of which were exclusively synthesized during phosphate starvation. When intact cells grown in the absence of phosphate were labelled with ¹²⁵I, several proteins were iodinated in addition of those observed in control cells, suggesting that the lack of phosphate induces some proteins located in the membranes or the periplasmic space of the bacteria. It is expected that by measuring the levels of expression of some of the proteins induced by the shortage of phosphorus, it might be possible to estimate in situ the relative physiological condition of the bacteria in a given bioleaching operation.     

溶磷微生物在土壤磷循环中的作用研究进展

磷是生物分子中的重要元素,是陆地生态系统初级生产的主要限制因子之一。全球粮食需求的增加和现代农业对磷肥的消耗导致集约农田中磷的过量输入,进而引起土壤磷流失的增加和地表水的持续富营养化。溶磷微生物(phosphate solubilizing microorganisms, PSMs)被认为是可以提高农业生产力的生态友好型肥料,在改善土壤肥力方面有重要意义。全面和深入理解PSMs功能及其在磷的土壤生物化学转化过程中的作用,对提高土壤磷有效性有至关重要的作用。本文系统综述了PSMs的种类和分布多样性,主要参与微生物磷循环的功能基因,以及PSMs如何参与土壤磷循环和这些过程背后的反应机制,以便更好地认识PSMs能力及其在土壤磷循环中的作用,以便于在未来的应用中发挥更大的潜力。     

Mathematical model of cell growth and phosphatase biosynthesis in Saccharomyces carlsbergensis under phosphate limitation.

The rate kinetics of growth and acid phosphate formation in the batch culture of Saccharomyces carlsbergensis LAM 1068 was studied under varying degrees of phosphate limitation. The mathematical model that was developed is concerned with the time lag for exponential growth, the biphasic growth on a substrate (glucose) and its product (ethanol), sustained growth on conservative phosphate, and the derepression of acid phosphatase. The numerical calculations using appropriate parametric constants successfully described the variation in the cell mass, glucose, ethanol, and inorganic phosphate concentrations, and the enzyme activity of acid phosphatase during aerobic growth of S. carlsbergensis under five different conditions of phosphate starvation. A simulation study revealed that the optimum initial phosphate concentration in the medium giving a high productivity of acid phosphatase was 2.0 mg phosphorus/g glucose liter.     

一株伯克霍尔德菌的筛选鉴定及溶磷性能优化

为解决我国大部分耕地土壤中可溶性磷含量不足、植物生长困难的问题,本研究对一株溶磷微生物(PB)进行了筛选鉴定及溶磷性能优化。结果表明: 菌株PB属于伯克霍尔德菌。该菌具有固氮和分泌吲哚-3-乙酸(IAA)等植物促生能力,对大肠杆菌也表现出一定的抑制效果;在pH 8.0～10.0范围内,菌株PB仍然能够保持较高的活性和溶磷能力,具有良好的耐碱性;溶磷性能优化结果表明,在30 ℃、pH 7.0、180 r·min^-1条件下,以葡萄糖为碳源、硫酸铵为氮源、磷酸三钙为磷源和添加50 μmol·L^-1赖氨酸时,菌株PB的溶磷能力达到最优,溶磷量为569.33 mg·L^-1,是优化前的1.9倍。该菌代谢过程中主要分泌柠檬酸、丙二酸和葡萄糖醛酸,添加赖氨酸后其分泌的有机酸种类不变,含量明显增加。盆栽试验表明,施用 PB菌肥能够显著促进大蒜幼苗的生长,而添加赖氨酸后促进效果更明显。与对照相比,PB加赖氨酸处理苗高增长18.6%,苗直径增长16.7%,地上部分鲜重和干重分别增长15.7%和22.1%,地下部分鲜重和干重分别增长22.0%和28.7%。土壤磷含量检测结果表明,PB和PB加赖氨酸处理土壤速效磷含量分别为对照的2.1和2.3倍,表明施用PB菌肥能够提高土壤中可溶性磷含量,而添加赖氨酸可以进一步强化PB菌肥的溶磷性能。     

Molecular cloning and functional analysis of a H+-dependent phosphate transporter gene from the ectomycorrhizal fungus Boletus edulis in southwest China

Phosphate transporters (PTs), as entry points for phosphorus (P) in organisms, are involved in a number of P nutrition processes such as phosphate uptake, transport, and transfer. In the study, a PT gene 1632 bp long (named BePT) was cloned, identified, and functionally characterized from Boletus edulis. BePT was expected to encode a polypeptide with 543 amino acid residues. The BePT polypeptide belonged to the major facilitator superfamily and showed a high degree of sequence identity to the Pht1 family. A topology model revealed that BePT exhibited 12 transmembrane helices, divided into two halves, and connected by a large hydrophilic loop in the middle. A yeast mutant complementation analysis suggested that BePT was a functional PT which mediated orthophosphate uptake of yeast at micromolar concentrations. Green fluorescent protein–BePT fusion proteins expressed were extensively restricted to the plasma membrane in BePT transformed yeast, and its activity was dependent on electrochemical membrane potential. In vitro, quantitative PCR confirmed that the expression of BePT was significantly upregulated at lower phosphorus availability, which may enhance phosphate uptake and transport under phosphate starvation. Our results suggest that BePT plays a key role in phosphate acquisition in the ectomycorrhizal fungus B. edulis.     

Environmental and physiological factors affecting the uptake of phosphate by Chlorobium limicola

The uptake of soluble phosphate by the green sulfur bacterium Chlorobium limicola UdG6040 was studied in batch culture and in continuous cultures operating at dilution rates of 0.042 or 0.064 h^–1. At higher dilution rates, washout occurred at phosphate concentrations below 7.1 μM. This concentration was reduced to 5.1 μM when lower dilution rates were used. The saturation constant for growth on phosphate (K _μ) was between 2.8 and 3.7 μM. The specific rates of phosphate uptake in continuous culture were fitted to a hyperbolic saturation model and yielded a maximum rate (Va _max) of 66 nmol P (mg protein)^–1 h^–1 and a saturation constant for transport (K _t) of 1.6 μM. In batch cultures specific rates of phosphate uptake up to 144 nmol P (mg protein)^–1 h^–1 were measured. This indicates a difference between the potential transport of cells and the utilization of soluble phosphate for growth, which results in a significant change in the specific phosphorus content. The phosphorus accumulated within the cells ranged from 0.4 to 1.1 μmol P (mg protein)^–1 depending on the growth conditions and the availability of external phosphate. Transport rates of phosphate increased in response to sudden increases in soluble phosphate, even in exponentially growing cultures. This is interpreted as an advantage that enables Chl. limicola to thrive in changing environments. Received: 9 February 1998 / Accepted: June 1998     

Growth optimization of the invasive cyanobacterium Cylindrospermopsis raciborskii in response to phosphate fluctuations

The role of ecophysiological traits in the success and expansion of the toxic cyanobacterium Cylindrospermopsis raciborskii is still under debate. One key factor appears to be the high physiological flexibility of this organism when obtaining limiting resources. Recent studies have found that filamentous bloom-forming cyanobacteria are able to optimize their growth by adjusting phosphate uptake during fluctuating nutrient conditions. We investigated the growth response of two phosphate-deficient C. raciborskii isolates (MVCC19 from Uruguay and CCMP1973 from USA) to short-term fluctuations in phosphate supply. These isolates were exposed to five phosphate concentrations which were provided in two supply modes: a single pulse (SingleP) versus the same amount divided in 10 pulses (TenP), with one pulse applied every 6 min. Morphological traits and changes in chlorophyll a and phycocyanin fluorescence were also evaluated. Growth rates of CCMP1973 and MVCC19 almost doubled and tripled, respectively, when exposed to multiple rather than single phosphate additions. Different growth rates were observed with the same total added resource, thus contradicting the classical model of dependence of growth rate on absolute external concentration. Phosphate-deficient C. raciborskii showed a remarkable physiological flexibility in adapting to phosphate availability on a timescale from minutes to hours. The TenP mode provided an extension of phosphate exposure time that allowed the energetic optimization of uptake and growth. The morphological plasticity of the species in response to phosphate supply mode was also shown by differences in trichome length and individual size. Our results are the first evidence of growth optimization of phosphate-deficient C. raciborskii to short-term nutrient fluctuations, revealing its physiological flexibility. This adaptive behaviour may help to explain the invasive success of this diazotrophic cyanobacterium in a wide range of aquatic ecosystems where phosphorus is frequently the limiting resource.     

OsCYCP4s coordinate phosphate starvation signaling with cell cycle progression in rice

Phosphate starvation leads to a strong reduction in shoot growth and yield in crops. The reduced shoot growth is caused by extensive gene expression reprogramming triggered by phosphate deficiency, which is not itself a direct consequence of low levels of shoot phosphorus. However, how phosphate starvation inhibits shoot growth in rice is still unclear. In this study, we determined the role of OsCYCP4s in the regulation of shoot growth in response to phosphate starvation in rice. We demonstrate that the expression levels of OsCYCP4s, except OsCYCP4;3, were induced by phosphate starvation. Overexpression of the phosphate starvation induced OsCYCP4s could compete with the other cyclins for the binding with cyclin‐dependent kinases, therefore suppressing growth by reducing cell proliferation. The phosphate starvation induced growth inhibition in the loss‐of‐function mutants cycp4;1, cycp4;2, and cycp4;4 is partially compromised. Furthermore, the expression of some phosphate starvation inducible genes is negatively modulated by these cyclins, which indicates that these OsCYCP4s may also be involved in phosphate starvation signaling. We conclude that phosphate starvation induced OsCYCP4s might coordinate phosphate starvation signaling and cell cycle progression under phosphate starvation stress.     

Distinctive features of dietary phosphate supply.

Dietary phosphate has profound effects on growth and renal handling of the compound. On the basis of changes in growth rate and food intake, after alterations in phosphate load, our laboratory previously suggested that these effects are mediated by intestinal signals (Landsman A, Lichtstein D, Bacaner M, and Ilani A. Br J Nutr 86: 217-223, 2001). The aim of this study was to further evaluate the role of dietary phosphate on food intake and appetite and specific organ growth, and to test for the presence of a serum factor that may affect renal phosphate handling in phosphate-resupplied rats. The experimental design was based on a comparison between groups of rats receiving identical low-phosphate diets but drinking water containing either phosphate or chloride. We show that 1) changes in food intake after alterations in phosphate load occurred in parallel with variations in digestive system distention, suggesting that dietary phosphate has also a direct effect on appetite; 2) dietary phosphate-dependent growth has a specific effect on the growth of liver and epididymal fat; and 3) serum of rats supplied with phosphate contains a factor that inhibits sodium-dependent phosphate transport in a model of renal proximal tubule cells. Collectively, these observations are in accord with the hypothesis that factor(s) emanating from the digestive system in response to dietary phosphate load may be involved in growth, appetite and renal handling of phosphate.