Importance of organic phosphate hydrolyzed in stalks of the lotic diatom <Emphasis Type="Italic">Didymosphenia geminata</Emphasis> and the possible impact of atmospheric and climatic changes

The aquatic colonial stalked diatom, Didymosphenia geminata, has acquired notoriety in recent years because of huge increases in many rivers of temperate regions. However, in some streams in northern England it has probably or, in the case of the R. Coquet (Northumberland), certainly been abundant for many decades. The paper describes the nutrient environment and phosphatase activities of Didymosphenia in Stony Gill (N. Yorkshire), a fast-flowing stream draining an upland catchment with peaty soils overlying limestone. Organic phosphate formed 85% of the filtrable phosphate in the water during the study (January–August 2000), with a maximum in April. Colonies were most abundant in June, but had disappeared by August. Surface phosphomonoesterase (PMEase) and phosphodiesterase activities assayed from March to July showed low PMEase activity in early March, but otherwise both were high throughout the period and especially so in June and July. Use of BCIP-NBT staining procedure showed that PMEase activity occurred in the stalks. A more detailed study of colony structure and staining with material from the R. Coquet in June 2006 also showed marked PMEase activity, with staining localized in the upper part of the stalks and the cells remaining unstained. It is suggested that organic phosphate is hydrolyzed in the stalk and the inorganic phosphate passes to the cell via a central tube in the stalk. It seems likely that organic phosphate as a major P source is a key factor favouring the success of Didymosphenia. The possible impact of environmental changes in the catchment such as climatic warming, C loss from peat and atmospheric N deposition is discussed. Handling editor: L. Naselli-Flores     

Colonization of biofilms by bacteria capable of biodegrading sodium dodecyl sulphate (SDS) at clean and polluted riverine sites

Fifty cyanobacterial strains (10 genera) were tested in batch culture for their ability to use organic phosphorus compounds (1 mg liter⁻¹ P) as their sole P source. Two monoesters, Na₂-β-glycerophosphate and π-nitrophenyl phosphate (πNPP), supported growth of all strains, and the diester bis-π-nitrophenyl phosphate (bis-π-NPP) and herring sperm DNA supported almost all strains. ATP was either a very favorable or poor P source and failed to support growth of nine strains, seven of which were Rivulariaceae with trichomes ending in a hair or long tapered region. Phytic acid was in general the least favorable P source. P-limited cultures grown initially with inorganic phosphate to conditions of P limitation were also tested for cell-bound and extracellular phosphomonoesterase (PMEase) and phosphodiesterase (PDEase) activities at two pH values (7.6, 10.3) using πNPP and bis-πNPP as substrates. Cell-bound PMEase was inducible in all strains and cell-bound PDEase in most strains. Most showed extracellular PMEase, but not extracellular PDEase. The highest values (μM πNPP or bis-πNPP hydrolyzed mg dry weight⁻¹ hour⁻¹) all occurred in strains ofGloeotrichia as follows: cell-bound PMEase at pH 7.6, 2.7 μM in strain D602; cell-bound PMEase at pH 10.3, 5.2 μM in D602; extracellular PMEase at pH 7.6, 0.73 μM in D281; extracellular PMEase at pH 10.3, 6.6 μM in D281; cell-bound PDEase at 7.6, 0.40 μM in D613; cell-bound PDEase at pH 10.3, 1.0 μM in D613. The results were compared to see if they indicated possible relationships between phosphatase activity and taxonomic or ecological grouping. The following differences were significant (P<0.05). Rivulariaceae produced higher yields than filamentous non-Rivulariaceae with β-glycerophosphate, πNPP, and DNA. Rivulariaceae with the ability to form hairs in culture showed poorer growth in ATP than non-hair-forming Rivulariaceae, but were more effective at utilizing phytic acid. Strains from calcareous environments had higher PMEase activity at pH 10.3 than strains from noncalcareous environments (P<0.01).     

Alkaline phosphatase activities of anAnabaena sp. from deep-water rice

Anabaena sp. grew with mono- and di-ester phosphate compounds as sources of phosphate, indicating the presence of phosphomonoesterase (PMEase) and phosphodiesterase (PDEase) activities. Cell-bound PMEase and PDEase activities were detected during growth in 0.5 and 10 mg PO₄l^–1 only when the cellular phosphate concentration fell to 0.46% of cell protein and the activities increased as cellular phosphate content decreased. The K_m values for these enzymes were 0.3mm forp-nitrophenyl phosphate and 0.2mm for bis-p-nitrophenyl phosphate, respectively. Only PMEase activity was found extracellularly. The pH optima for PMEase and PDEase were 10.2 and 10.4, respectively, and the temperature optima at pH 10.2 were 37°C and 40°C, respectively. Ca²⁺ increased the enzyme activities while Zn²⁺ caused marked inhibition. The inorganic phosphate repressed the cellular PMEase activity after a lag of 4 h.     

Factors modulating alkaline phosphatase activity in the diazotrophic rice-field cyanobacterium, Anabaena oryzae

Summary Alkaline phosphatases (APase), both phosphomonoesterase (PMEase) and phosphodiesterase (PDEase) were studied in the cyanobacterium Anabaena oryzae for their specific requirements of temperature, pH, micro- and macronutrients and their activities in the presence of salinity and heavy metal stress. The alkaline phosphatases (PMEase and PDEase) are quite stable enzymes and require a narrow range of pH (pH 10–10.2) and temperature (35–40 °C) for their optimal activity.A pH of 10, 10.2 and 10.2 supported optimal activity of cellular PMEase, cellular PDEase and extracellular PMEase, respectively, whereas temperatures of 35, 38 and 40 °C were required for their optimal activity. The requirement for Ca²⁺ and Mg²⁺ as macronutrients and the significance of the micronutrients Zn²⁺, Co²⁺, Fe²⁺, Mn²⁺ and Cu²⁺ in APase activity in the cyanobacterium suggests nutritional regulation of enzyme activity in A. oryzae. The metals Pb²⁺, Cr⁶⁺ and Ni²⁺ severely inhibited APase activity, whereas the NaCl stress had a dual role, which was concentration dependent. NaCl stress at lower concentrations (≤20 mM) caused an increase in cellular PMEase activity while its higher concentration (>20 mM) favoured release of the extracellular PMEase. The decrease in cellular activity and an increase in extracellular activity suggest that the higher concentrations of salt stimulate the release of the enzyme.The data suggest that the cyanobacterium A. oryzae possess a potential application as biofertilizer in high salinity and alkaline (Ca²⁺-rich) soils because of its ability to release PO₄³⁻ enzymatically under these conditions.     

Biology of the freshwater diatom <Emphasis Type="Italic">Didymosphenia</Emphasis>: a review

Features of the colonial diatom Didymosphenia are reviewed, especially D. geminata. Although there is a long record of its occurrence in north temperate regions, mass growths have been reported much more widely in recent years. Contrary to some statements in the literature, there are also reliable older records for the southern hemisphere, though the first report of mass growth was in New Zealand in 2004. The annual cycle of morphological changes in D. geminata in northern England, and probably elsewhere, includes a winter period when motile cells are attached to the substratum followed by spring when stalks start to develop. These raise cells into the water column and provide a site for phosphatase activity. Environmental factors associated with success include absence of extreme floods, high light, pH above neutral and nutrient chemistry. D. geminata often, but not always, occurs in waters where the N:P ratio is high for much of the year, but the key factor is the ratio of organic to inorganic phosphate. D. geminata thrives where organic P is predominant and the overall P concentration is low enough for organic P to be an important P source. It is unknown whether organic N can be used. Environmental changes increasing the relative importance of organic P are likely to favour D. geminata. Likely examples are increased N:P due to atmospheric N deposition and changes in form and seasonality of P release from organic-rich soils due to climatic warming. The nutrient chemistry of deep water released from dams to rivers also needs investigation. To what extent are genetic changes occurring in response to environmental changes and are new ecotypes spreading round the world? In spite of many adverse reports about D. geminata, such as detached mats blocking water pipes, there is still doubt about the extent to which it causes problems, particularly for fish. There have been few adverse effects on migratory salmonids in Europe and North America, but at least one report of harm to a brown trout population in USA. In New Zealand, it has caused serious problems for water sports, although it remains open to question how much adverse effect it has had on fish populations. If the presence of microcystins in or associated with D. geminata, as indicated recently for two populations, proves to be widespread and at sufficiently high concentration, their possible accumulation in fish requires study. Where control is required, this could be achieved by enhancing the ratio of inorganic to organic phosphate in the water early in the growth season. Practical ways to achieve this are suggested.     

Seasonal changes in the surface phosphatase kinetics of aquatic mosses in northern England

Abstract

The widespread occurrence of surface phosphatase activity in aquatic mosses, presumably synthesized to utilize organic phosphates in the environment, suggests the likelihood of strategies to optimize this process. Here we report seasonal changes in the kinetics of surface phosphatase in six moss populations, representing four species (Fontinalis antipyretica, Fontinalis squamosa, Rhynchostegium riparioides and Warnstorfia fluitans) from N.E. England. Kinetic parameters of shoots assayed within a few hours of sampling from the streams showed a summer/autumn increase in K _m and V _max corresponding to seasonal decrease in phosphate supply. The detection of phosphomonoesterases and phosphodiesterases with both low and high substrate affinities in all the populations studied confirms the widespread existence of this phenomenon in mosses. Similar two-phase kinetics were found for clonal, axenic isolates of two of the species (Rhynchostegium and Warnstorfia), indicating that any influence of epiphytes on the results with stream material was negligible. The seasonal changes in the kinetic parameters indicate that surface phosphatase activity is a dynamic response to seasonal changes in nutrient supply and/or requirement.     

Arbuscular mycorrhizal fungi and organic fertilizer influence photosynthesis,root phosphatase activity,nutrition, and growth of <Emphasis Type="Italic">Ipomoea carnea</Emphasis> ssp. <Emphasis Type="Italic">fistulosa</Emphasis>

The effect of arbuscular mycorrhizal fungi (AMF) inoculation and organic slow release fertilizer (OSRF) on photosynthesis, root phosphatase activity, nutrient acquisition, and growth of Ipomoea carnea N. von Jacquin ssp. fistulosa (K. Von Martinus ex J. Choisy) D. Austin (bush morning glory) was determined in a greenhouse study. The AMF treatments consisted of a commercial isolate of Glomus intraradices and a non-colonized (NonAMF) control. The OSRF was applied at 10, 30, and 100 % of the manufacturer’s recommended rate. AMF plants had a higher net photosynthetic rate (P _N), higher leaf elemental N, P, and K, and generally greater growth than NonAMF plants. Total colonization levels of AMF plants ranged from 27 % (100 % OSRF) to 79 % (30 % OSRF). Root acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were generally higher in AMF than non-AMF plants. When compared to NonAMF at 100 % OSRF, AMF plants at 30 % OSRF had higher or comparable ACP and ALP activity, higher leaf elemental P, N, Fe, Cu, and Zn, and a greater P _N (at the end of the experiment), leading to generally greater growth parameters with the lower fertility in AMF plants. We suggest that AMF increased nutrient acquisition from an organic fertilizer source by enhancing ACP and ALP activity thus facilitating P acquisition, increasing photosynthesis, and improving plant growth.     

Utilization of phosphate diester by the marine diatom Chaetoceros ceratosporus

Utilization of phosphate diester (PDE) and phosphodiesterase(PDEase) production by five marine phytoplankton species wereexamined in the laboratory to evaluate the contribution of PDEto the growth of phytoplankton. Among the five marine phytoplanktonspecies tested, only Chaetoceros ceratosporus was able to usethe PDE compound, bis(p-nitrophenyl) phosphate (bis-NPP), effectivelyas a sole phosphorus source. In addition, C. ceratosporus simultaneouslyproduced both PDEase and alkaline phosphatase (APase) at almostequal activity levels under the phosphate-deficient condition.These results suggest that PDE compounds presumably play animportant role as a phosphorus source for PDEase-producing phytoplanktonin coastal environments.     

Phosphorus source alters host plant response to ectomycorrhizal diversity

We examined the influence of phosphorus source and availability on host plant (Pinus rigida) response to ectomycorrhizal diversity under contrasting P conditions. An ectomycorrhizal richness gradient was established with equimolar P supplied as either inorganic phosphate or organic inositol hexaphosphate. We measured growth and N and P uptake of individual P. rigida seedlings inoculated with one, two, or four species of ectomycorrhizal fungi simultaneously and without mycorrhizas in axenic culture. Whereas colonization of P. rigida by individual species of ectomycorrhizal fungi decreased with increasing fungal richness, colonization of all species combined increased. Plant biomass and N content increased across the ectomycorrhizal richness gradient in the organic but not the inorganic P treatment. Plants grown under organic P conditions had higher N concentration than those grown under inorganic P conditions, but there was no effect of richness. Phosphorus content of plants grown in the organic P treatment increased with increasing ectomycorrhizal richness, but there was no response in the inorganic P treatment. Phosphorus concentration was higher in plants grown at the four-species richness level in the organic P treatment, but there was no effect of diversity under inorganic P conditions. Overall, few ectomycorrhizal composition effects were found on plant growth or nutrient status. Phosphatase activities of individual ectomycorrhizal fungi differed under organic P conditions, but there was no difference in total root system phosphatase expression between the inorganic or organic P treatments or across richness levels. Our results provide evidence that plant response to ectomycorrhizal diversity is dependent on the source and availability of P.     

Effect of Substrate and Cell Surface Hydrophobicity on Phosphate Utilization in Bacteria

We measured the rates of utilization of hydrophobic and hydrophilic phosphate compounds in gram-negative bacteria with different surface hydrophobicities, isolated from wetland habitats. Three hydrophobic and two hydrophilic bacterial species were selected for study by measuring cell adherence to hydrocarbons. The bacteria were grown under phosphorus-limited conditions with P(infi), hydrophilic (beta)-glycerophosphate, or hydrophobic phosphatidic acid as the phosphate source. Hydrophilic bacteria grew most rapidly on P(infi), followed by (beta)-glycerophosphate. Phosphatidic acid did not support growth or did so at a much later time (40 h) than did the other phosphate treatments. Although all hydrophobic species grew well on these substrates, the rate of growth of two Acinetobacter baumannii isolates on phosphatidic acid exceeded the rate of growth on phosphate or (beta)-glycerophosphate. A membrane phospholipid and lipopolysaccharide were used as a source of phosphorus by hydrophobic species, whereas hydrophilic species could not use the membrane phospholipids and used lipopolysaccharide to a lesser extent. Besides hydrophobic interaction between cells and substrate, phosphatase activity, which was cell bound in hydrophilic species but 30 to 50% unbound in hydrophobic species, affected cell growth. Dialyzed culture supernatant containing phosphatase from hydrophobic species increased the phosphate availability to hydrophilic species. Additionally, cellular extracts from a hydrophilic species, when added to hydrophilic cells, permitted growth on hydrophobic phosphate sources. Naturally occurring amphiphilic humic acids affected the utilization of P(infi) and (beta)-glycerophosphate in bacteria with hydrophilic surfaces but did not affect hydrophobic bacteria. Our results indicate that hydrophobic phosphate sources can be used by bacteria isolated from aquatic environments as the sole phosphorus source for growth. This utilization, in part, appears to be related to cell surface hydrophobicity and extracellular enzyme production.     

Soil Phosphorus Availability and Transformation Rates in Relictic Pinsapo Fir Forests from Southern Spain

Abies pinsapo fir forests are remnant of temperate-like coniferous forests currently subjected to the typical seasonal constraints of Mediterranean-type climates. We have examined for the first time, P availability and cycling in A. pinsapo forests from southern Spain by using both, measurements of concentrations of soil P fractions (resin extractable P, bicarbonate extractable P and NaOH extractable P) and rates of P supply to (P solubilization, P mineralization and phosphomonoesterases (PMEase activity) and P immobilization from the soil solution. Soils from A. pinsapo stands differing in lithology (serpentinitic and calcareous substrates) and successional status (from young-agradative to old-growth stages) were chosen for this study. Labile organic P fractions, PMEase activity and rates of P gross mineralization and immobilization were significantly higher in agradative stands on serpentines than in successionally comparable calcareous stands. This suggests an important role of the organic P subcycle in the A. pinsapo stand on serpentine. On calcareous lithology, PMEase activity and all soil P transformation rates significantly increased throughout the successional series. Similarly, all organic and inorganic P fractions measured in old-growth forests showed the maximum values of the series. These trends fit the predictions of standard patterns of P cycling changes along with succession, in which P supply to plants greatly depends on solubilization from mineral forms at early-to-mid successional stages, whereas the importance of processes related to the organic P subcycle increases as succession progresses.     

Variation in root-associated phosphatase activities in wheat contributes to the utilization of organic P substrates in vitro,but does not explain differences in the P-nutrition of plants when grown in soils

To understand whether genotypic variation in root-associated phosphatase activities in wheat impacts on its ability to acquire phosphorus (P), various phosphatase activities of roots were measured in relation to the utilization of organic P substrates in agar, and the P-nutrition of plants was investigated in a range of soils. Root-associated phosphatase activities of plants grown in hydroponics were measured against different organic P substrates. Representative genotypes were then grown in both agar culture and in soils with differing organic P contents and plant biomass and P uptake were determined. Differences in the activities of both root-associated and exuded phosphodiesterase and phosphomonoesterase were observed, and were related to the P content of plants supplied with either ribonucleic acid or glucose 6-phosphate, respectively, as the sole form of P. When the cereal lines were grown in different soils, however, there was little relationship between any root-associated phosphatase activity and plant P uptake. This indicates that despite differences in phosphatase activities of cereal roots, such variability appears to play no significant role in the P-nutrition of the plant grown in soil, and that any benefit derived from the hydrolysis of soil organic P is common to all genotypes.     

Variation in root-associated phosphatase activities in wheat contributes to the utilization of organic P substrates in vitro,but does not explain differences in the P-nutrition of plants when grown in soils

To understand whether genotypic variation in root-associated phosphatase activities in wheat impacts on its ability to acquire phosphorus (P), various phosphatase activities of roots were measured in relation to the utilization of organic P substrates in agar, and the P-nutrition of plants was investigated in a range of soils. Root-associated phosphatase activities of plants grown in hydroponics were measured against different organic P substrates. Representative genotypes were then grown in both agar culture and in soils with differing organic P contents and plant biomass and P uptake were determined. Differences in the activities of both root-associated and exuded phosphodiesterase and phosphomonoesterase were observed, and were related to the P content of plants supplied with either ribonucleic acid or glucose 6-phosphate, respectively, as the sole form of P. When the cereal lines were grown in different soils, however, there was little relationship between any root-associated phosphatase activity and plant P uptake. This indicates that despite differences in phosphatase activities of cereal roots, such variability appears to play no significant role in the P-nutrition of the plant grown in soil, and that any benefit derived from the hydrolysis of soil organic P is common to all genotypes.     

秦岭地区华北落叶松人工林地土壤养分和酶活性变化

以秦岭地区不同林龄(5年生、10年生、20年生、30年生和40年生)华北落叶松人工林为研究对象,采用野外调查采样和室内分析相结合的方法,研究了不同林龄华北落叶松人工林地土壤剖面p H值、有机质养分和酶活性的变化。结果表明:土壤p H值随着林龄有降低趋势,随着土层深度的增加有升高趋势。土壤有机质和土壤速效氮、速效磷和速效钾在近熟龄期显著高于幼龄期。土壤中的磷酸酶、脲酶、蔗糖酶和过氧化氢酶活性随着林龄都呈"高—低—高"的趋势,幼龄期的土壤蔗糖酶活性显著高于近熟龄的,而磷酸酶、脲酶和过氧化氢酶活性是近熟龄期的显著高于中幼龄。土壤速效养分和酶活性都随着土层深度的增加有显著的降低。相关分析表明磷酸酶活性与有机质、速效氮、速效磷、速效钾和脲酶活性呈极显著的正相关性(P0.01),脲酶与有机质、速效氮和速效钾呈极显著正相关。蔗糖酶与过氧化氢酶活性显著负相关(P0.05),与p H值有一定的正相关性。秦岭地区华北落叶松人工林进入近熟林之后土壤肥力有一定的恢复,而在中幼龄阶段土壤养分比较缺乏,尤其是氮磷。     

Regulation of dauciform root formation and root phosphatase activities of sedges (<Emphasis Type="Italic">Carex</Emphasis>) by nitrogen and phosphorus

Aims

Dauciform roots (DR) are formed by some Cyperaceae under phosphorus (P) deficiency. To advance our understanding of their physiological function, I ask: Is DR formation regulated by shoot P status or external P supply? How does it respond to nitrogen (N)? Do DR enhance root monoesterase, diesterase or phytase activities and ability to utilize organic P?

Methods

Greenhouse experiments were carried out with two Carex species grown in sand with (1) different combinations of N and P supply, (2) local supply of N or P to root halves, and (3) different organic P forms.

Results

Carex flava produced DR in all treatments. The density of DR and phosphatase activities increased with N supply; they were regulated by shoot P status and external N (but not P) supply. All phosphatase activities increased with DR density. Carex muricata produced no DR and had lower diesterase activity than C. flava but both species grew equally well with diester-P.

Conclusions

DR and phosphatase activities are regulated by both N and P supply. Similar growth responses to nutrients in both species suggest small costs and benefits of DR under experimental conditions but confirmation is needed for plants grown on natural soils.

     

Dependence of the maximum temperature for growth of Saccharomyces cerevisiae on nutrient concentration

Anacystis nidulans (Synechococcus) was maintained in a medium of low phosphate concentration (0.1 mM) and grew with a normal doubling time of 5 hrs at 30°C. Such cultures ahd a normal pigment composition and alkaline phosphatase was detectable at low specific activities only.The onset of phosphate-limited growth occurred when the phosphate concentration in the medium fell to a value below 4 M (the limit of accurate determination by the assay method used) and resulted in increases in alkaline phosphatase activity, reaching a final 10 to 15 fold increase in specific activity after a period of several hours. Marked changes in the overall pigment composition occurred in this period of growth restriction. The addition of phosphate to such cultures resulted in a halt in synthesis of the enzyme and the restoration of normal pigmentation before growth resumed at the normal rate.Several organic phosphate esters could replace inorganic phosphate for growth and were also hydrolyzed by the partially purified enzyme, but growth rates were characteristically lower and the specific activity only 3 to 4 fold higher than in cultures grown in phosphate excess.Studies with the partially purified enzyme suggested that it differed in some of its properties from other alkaline phosphatases described in the literature.Abbreviations Used pNP pnitrophenol - pNPP pnitrophenylphosphate     

Soil Phosphorus Uptake by Continuously Cropped Lupinus albus: A New Microcosm Design

When grown in soils with sparingly available phosphorus (P), white lupin (Lupinus albus L.) forms special root structures, called cluster roots, which secrete large amounts of organic acids and concomitantly acidify the rhizosphere. Many studies dealing with the understanding of this P acquisition strategy have been performed in short time experiments either in hydroponic cultures or in small microcosm designs with sand or sand:soil mixtures. In the present study, we applied an experimental design which came nearer to the natural field conditions: we performed a one-year experiment on large microcosms containing 7 kg of soil and allowing separation of rhizosphere soil and bulk soil. We planted six successive generations of lupins and analysed P uptake, organic P desorption, phosphatase activities and organic acid concentrations in different soil samples along a spatio-temporal gradient. We compared the rhizosphere soil samples of cluster (RSC) and non-cluster roots (RSNC) as well as the bulk soil (BS) samples. A total shoot biomass of 55.69 ± 1.51 g (d.w.) y⁻¹ was produced and P uptake reached 220.59 ± 5.99 mg y⁻¹. More P was desorbed from RSC than from RSNC or BS (P < 0.05). RSC and RSNC showed a higher activity of acid and alkaline phosphatases than BS samples and a higher acid phosphatase activity was observed in RSC than in RSNC throughout the one-year experiment. Fumarate was the most abundant organic acid in all rhizosphere soil samples. Citrate was only present in detectable amounts in RSC while malate and fumarate were recovered from both RSC and RSNC. Almost no organic acids could be detected in the BS samples. Our results demonstrated that over a one-year cultivation period in the absence of an external P supply, white lupin was able to acquire phosphate from the soil and that the processes leading to this P uptake took place preferentially in the rhizosphere of cluster roots.     

Mycelial forms of <Emphasis Type="Italic">Pseudallescheria boydii</Emphasis> present ectophosphatase activities

Phosphatase activities were characterized in intact mycelial forms of Pseudallescheria boydii, which are able to hydrolyze the artificial substrate p-nitrophenylphosphate (p-NPP) to p-nitrophenol (p-NP) at a rate of 41.41 ± 2.33 nmol p-NP per h per mg dry weight, linearly with increasing time and with increasing cell density. MgCl₂, MnCl₂ and ZnCl₂ were able to increase the (p-NPP) hydrolysis while CdCl₂ and CuCl₂ inhibited it. The (p-NPP) hydrolysis was enhanced by increasing pH values (2.5-8.5) over an approximately 5-fold range. High sensitivity to specific inhibitors of alkaline and acid phosphatases suggests the presence of both acid and alkaline phosphatase activities on P. boydii mycelia surface. Cytochemical localization of the acid and alkaline phosphatase showed electron-dense cerium phosphate deposits on the cell wall, as visualized by electron microscopy. The product of p-NPP hydrolysis, inorganic phosphate (Pi), and different inhibitors for phosphatase activities inhibited p-NPP hydrolysis in a dose-dependent manner, but only the inhibition promoted by sodium orthovanadate and ammonium molybdate is irreversible. Intact mycelial forms of P. boydii are also able to hydrolyze phosphoaminoacids with different specificity.     

Inorganic phosphate transport in Escherichia coli: involvement of two genes which play a role in alkaline phosphatase regulation

Two classes of alkaline phosphatase constitutive mutations which comprise the original phoS locus (genes phoS and phoT) on the Escherichia coli genome have been implicated in the regulation of alkaline phosphatase synthesis. When these mutations were introduced into a strain dependent on a single system, the pst system, for inorganic phosphate (P(i)) transport, profound changes in P(i) transport were observed. The phoT mutations led to a complete P(i) (-) phenotype in this background, and no activity of the pst system could be detected. The introduction of the phoS mutations changed the specificity of the pst system so that arsenate became growth inhibitory. Changes in the phosphate source led to changes in the levels of constitutive alkaline phosphatase synthesis found in phoS and phoT mutants. When glucose-6-phosphate or l-alpha-glycerophosphate was supplied as the sole source of phosphate, phoT mutants showed a 3- to 15- fold reduction in constitutive alkaline phosphatase synthesis when compared to the maximal levels found in limiting P(i) media. However, these levels were still 100 times greater than the basal level of alkaline phosphatase synthesized in wild-type strains under these conditions. The phoS mutants showed only a two- to threefold reduction when grown with organic phosphate sources. The properties of the phoT mutants selected on the basis of constitutive alkaline phosphatase synthesis were similar in many respects to those of pst mutants selected for resistance to growth inhibition caused by arsenate. It is suggested that the phoS and phoT genes are primarily involved in P(i) transport and, as a result of this function, play a role in the regulation of alkaline phosphatase synthesis.     

Effect of Phosphorus Deficiency and Water Deficit on Phosphatase Activities from Wheat Leaves

Wheat was sown in a phosphorus (P) deficient soil. Plants atlow levels of applied P had lower growth rates and lower concentrationsof phosphate in the shoots than plants grown with ‘highP’. Activities of both insoluble and soluble phosphataseincreased with P deficiency in the mature leaves. Soluble phosphataseactivities increased 2.5–3.0 fold as the concentrationof phosphate in the leaves fell from 0.4% to 0.1% dry weightThis increase was not a consequence of reduced growth, as severenitrogen deficiency had no effect on phosphatase activity. Soluble phosphatase activities were higher in young than inmature leaves, and also increased 3–4 fold with severewater deficit. However these increases in activity were notaccompanied by low concentrations of phosphate. Moreover, solublephosphatase activities in mature leaves of plants grown underconditions of water deficit rapidly decreased after rewatering.In contrast, the high soluble phosphatase activities in matureleaves of P deficient wheat persisted for up to 12 d after theresupply of P to adequate levels.