Effect of temperature and pH on absorption of carbon dioxide by a free level of mixed solutions of some buffers

The rate of absorption of carbon dioxide by solutions of NaHCO₃, KH₂PO₄, hydrogencarbonate, phosphate and borate buffers at 20, 30 and 40°C was determined manometrically. The absorption rate increases for all buffers tested with increasing pH. The CO₂ absorption rate by KH₂PO₄ and by the phosphate buffer at low pH is lower than that of water. For other buffers tested it is equal to or higher than that of water, especially at higher temperatures.     

The Influence of Phosphate Uptake on Cation Uptake in Fusarium oxysporum f. sp. vasinfectum

Fusarium oxysporum grown in a low phosphate medium was found to take up several times as much K from KH₂PO₄ as from KCI solutions. Large amounts of phosphate also were taken up from KH₂PO₄. Similar large uptakes of Na and phosphate took place from solutions of NaH₂PO₄. Substantial quanties of phosphate were taken up from solutions of Ca(H₂PO₄)₂ in the absence of any appreciable Ca uptake. When the fungus was grown in a medium containing high phosphate, little or no uptake of phosphate from KH₂PO₄ solutions occured and the K Uptake was at the same level as from KCI solutions. During large phosphate uptake sizable reductions in the organic acid content of the fungal cells were observed. Much, but not all, of the data could be explained on the basis of maintenance of charge balance within the cells. – The respiratory rate of fungus, grown in a low P medium, was markedly increased in KH₂PO₄ solution. Fungus, grown in a medium with high phosphate, had a higher respiratory rate which showed only a slight response to KH₂PO₄ solution. Fungus, grown in a medium with high phosphate, had a higher respiratory rate which showed only a slight response to KH₂PO₄.     

Development and application of a nutrient-diffusing bioassay for large rivers

1. Laboratory and field experiments were performed to develop and then apply a nutrient-diffusing substratum (NDS) design suitable for use in large, fast-flowing rivers. 2. Initial laboratory experiments quantified diffusion of PO₄ and NO₃ from new and previously used clay pots, which were soaked in deionized distilled water. Mean release rates initially exceeded 2.4 and 725 μmol l^–1 day^–1 P and 0.22 and 18 μmol l^–1 day^–1 N from new and used pots, respectively, but declined rapidly with increasing time spent in deionized distilled water and were below detectable levels after about 18 and 29 days, respectively. 3. A phosphorus (P) dose–response experiment in a P-limited reach of the Athabasca River, Alberta, Canada showed that epilithic biomass and macroinvertebrate density on NDS increased with increasing concentrations of KH₂PO₄ up to about 0.5 m . Beyond this threshold, biomasses and densities were unaffected by initial KH₂PO₄ concentration. Coefficients of variation of epilithic biomass estimates declined with increasing KH₂PO₄ whereas invertebrate density appeared to be unaffected by KH₂PO₄ levels. 4. Release rates of both P and N from NDS filled with 0.5 m KH₂PO₄ or 0.5 m NaNO₃ declined at a log-negative rate from about 5000 μmol N-NO₃ l^–1 day^–1 and 3500 μmol P-PO₄ l^–1 day^–1 on day 2, to 200 μmol l^–1 day^–1 for both N and P on day 32. 5. After development, we used the diffusing substrata to identify spatial patterns in nutrient limitation at seven sites along a 120 km reach in the Athabasca River, that receives two known point-source nutrient inputs. NDS consisting of N, P, N + P and unenriched controls were attached to the river bottom for 22–23 days and then retrieved and sampled for epilithic chlorophyll a. Physicochemical parameters and epilithic biomasses on upper stone surfaces were also quantified when NDS were deployed and retrieved from each site. 6. Sites located immediately downstream of the two point source inputs had higher water column concentrations of PO₄ and epilithic biomasses than the site immediately upstream; epilithic biomass was positively related to PO₄ in the late autumn (r²⁼ 0.58) but not in early autumn. Sites located immediately below nutrient inputs were not nutrient-limited, whereas upstream reference sites were P-limited.     

Effects of Foliar Sprays of Phosphates on Powdery Mildew (Sphaerotheca pannosa) of Roses

Powdery mildew on plants of a local clone of Rosa indica Major was significantly controlled by a single spray of 25 mM aqueous solutions of K₂HPO₄, KH₂PO₄ plus KOH, or NaHCO₃, all plus Tween 20 (0.5 ml/l) or bupirimate (Nimrod) at 0.5 ml/l, which was applied 4 days before inoculation with conidial suspension of Sphaerotheca pannosa var. rosae. Disease incidence was reduced by 79, 71, 54 and 50%, as compared to controls on plants sprayed with KH₂PO₄ plus KOH, K₂HPO₄, NaHCO₃ or bupirimate, respectively. Phosphates were suppressive and expressed by disappearance of 99% of the pustules and conidia, as early as 2 days after a single spray on mildewed foliage. This treatment was efficient for at least 9 days after the first application when large infected greenhouse-grown plants were used. Re-application of these salts on the same plants reduced the lesion area by about 90% from that recorded before the application. Phosphate and bicarbonate were more suppressive than the systemic fungicide bupirimate in the early period (up to 2 days). The suppresion effects of bicarbonate and bupirimate, however, were short-term and not persistent, while the phosphate treatments remained significantly suppressive for up to 23 days, when the experiment was terminated. The inhibitory and suppressive effectiveness of phosphate salts is discussed in the light of their possible acceptance as ideal foliar fertilizers which should be considered for use in the field for disease control.     

Effects of Foliar Sprays of Phosphates on Powdery Mildew (Sphaerotheca pannosa) of Roses

Powdery mildew on plants of a local clone of Rosa indica Major was significantly controlled by a single spray of 25 mM aqueous solutions of K₂HPO₄, KH₂PO₄ plus KOH, or NaHCO₃, all plus Tween 20 (0.5 ml/l) or bupirimate (Nimrod) at 0.5 ml/l, which was applied 4 days before inoculation with conidial suspension of Sphaerotheca pannosa var. rosae. Disease incidence was reduced by 79, 71, 54 and 50%, as compared to controls on plants sprayed with KH₂PO₄ plus KOH, K₂HPO₄, NaHCO₃ or bupirimate, respectively. Phosphates were suppressive and expressed by disappearance of 99% of the pustules and conidia, as early as 2 days after a single spray on mildewed foliage. This treatment was efficient for at least 9 days after the first application when large infected greenhouse-grown plants were used. Re-application of these salts on the same plants reduced the lesion area by about 90% from that recorded before the application. Phosphate and bicarbonate were more suppressive than the systemic fungicide bupirimate in the early period (up to 2 days). The suppresion effects of bicarbonate and bupirimate, however, were short-term and not persistent, while the phosphate treatments remained significantly suppressive for up to 23 days, when the experiment was terminated. The inhibitory and suppressive effectiveness of phosphate salts is discussed in the light of their possible acceptance as ideal foliar fertilizers which should be considered for use in the field for disease control.     

A comparison of different chemicals for osmotic treatment of vegetable seed

Samples of carrot, celery, leek and onion seed were treated before sowing by imbibition in osmotic solutions of polyethylene glycol 6000 (PEG), glycerol and potassium dihydrogen orthophosphate (KH₂PO₄). The solutions were sufficiently concentrated to prevent germination during treatment, and within each species, the amount of water taken up by the seeds during treatment did not vary greatly between solutions. All treatments increased the rates of seed germination and seedling emergence compared to untreated controls, but glycerol-treated seeds germinated and emerged significantly more slowly than did PEG- and KH₂PO₄-treated seeds. The effects of PEG and glycerol treatments on percentage germination and seedling emergence were small, but KH₂PO₄ treatment tended to reduce percentage germination and emergence, most markedly in leek and one cultivar of celery. It is concluded that PEG treatment gives the most consistently beneficial effects for the range of species tested.     

Extractable sulphate and organic sulphur in soils and their availability to plants

Ten soils collected from the major arable areas in Britain were used to assess the availability of soil sulphur (S) to spring wheat in a pot experiment. Soils were extracted with various reagents and the extractable inorganic SO₄-S and total soluble S(SO₄-S plus a fraction of organic S) were determined using ion chromatography (IC) or inductively-coupled plasma atomic emission spectrometry (ICP-AES), respectively. Water, 0.016 M KH₂PO₄, 0.01 M CaCl₂ and 0.01 M Ca(H₂PO₄)₂ extracted similar amounts of SO₄-S, as measured by IC, which were consistently smaller than the total extractable S as measured by ICP-AES. The amounts of organic S extracted varied widely between different extractants, with 0.5 M NaHCO₃ (pH 8.5) giving the largest amounts and 0.01 M CaCl₂ the least. Organic S accounted for approximately 30–60% of total S extracted with 0.016 M KH₂PO₄ and the organic C:S ratios in this extract varied typically between 50 and 70. The concentrations of this S fraction decreased in all soils without added S after two months growth of spring wheat, indicating a release of organic S through mineralisation. All methods tested except 0.5 M NaHCO₃-ICP-AES produced satisfactory results in the regression with plant dry matter response and S uptake in the pot experiment. In general, 0.016 M KH₂PO₄ appeared to be the best extractant and this extraction followed by ICP-AES determination was considered to be a good method to standardise on.     

Enhanced curdlan production in <Emphasis Type="Italic">Agrobacterium</Emphasis> sp. ATCC 31749 by addition of low-polyphosphates

A large amount of adenosine triphosphate with high energy phosphate bonds is required for uridine triphosphate regeneration during curdlan biosynthesis by Agrobacterium sp. ATCC 31749. To supply high energy for curdlan synthesis, three low-polyphosphates (Na₄P₂O₇, Na₅P₃O₁₀, and (NaPO₃)₆) with higher energy phosphate bonds were employed to substitute for KH₂PO₄-K₂HPO₄ in fermentation medium. Two genes encoding the polyphosphate metabolizing enzymes, polyphosphate kinase and exopolyphosphatase, were amplified and showed 95% homology to those in Agrobacterium sp. C58 by sequence analysis. The curdlan yields were enhanced by 23 and 134% when phosphate concentrations 0.024 mol/L of Na₅P₃O₁₀ and 0.048 mol/L of (NaPO₃)₆ respectively, were added in the medium. The maximum curdlan yield of 30 ± 1.02 g/L was obtained with the addition of 0.048 mol/L of (NaPO₃)₆ with 5 g/L CaCO₃ in the medium. When CaCO₃ was removed from the culture and the three lowpolyphosphates were added, the pH and biomass yield dropped remarkably and little or no curdlan was produced. The culture containing 0.048 mol/L of (NaPO₃)₆ was mixed with KH₂PO₄-K₂HPO₄ and CaCO₃ in the medium, but showed no effect on curdlan production. However, curdlan yield was improved by 49 ∼ 60% when CaCO₃ was removed from the medium and KH₂PO₄-K₂HPO₄ acted as a buffer. It appears that the positive effect of (NaPO₃)₆ on curdlan production required the buffering capacity of CaCO₃ and the absence of KH₂PO₄-K₂HPO₄ competing as a phosphate supplier.     

Studies on isolated cell components: I. Nuclear isolation by differential centrifugation

A method is presented for isolating nuclei of rat and hamster liver in a high state of purity and in a condition optically similar to nuclei within living cells.The isolation procedure consists in the homogenization and differential centrifugation of fresh liver at 0–5 ° in a salt-sucrose solution buffered at pH 7.1. By layering the material to be centrifuged over a relatively large volume of a slightly denser solution the purification can be carried out in 4 or 5 centrifugations. The entire procedure can be completed in about 90 minutes. The yield as determined by measurements of desoxyribonucleic acid is about 5 per cent.The solutions contain KH₂PO₄, K₂HPO₄, NaHCO₃, and sucrose. The sucrose concentration is varied to give density differences required for layering. Salt-sucrose solutions maintain a large proportion of the isolated nuclei in a nongranular condition for 6 hours at 0 °. Pure sucrose is less satisfactory for maintenance.The protein-desoxyribonucleic acid ratio for the isolated nuclei averages 5.1 with a range of 2.7 to 8.9.     

Aqueous two-phase extraction coupled with ultrafiltration for purification of amyloglucosidase

Aqueous two phase extraction (ATPE) in combination with ultrafiltration was employed for concentration and purification of amyloglucosidase produced by solid state fermentation. After extraction (with water) from dry moldy bran the dilute enzyme extract was concentrated by ATPE in a polyethylene glycol (PEG)/maltodextrin (MDX) system. The enzyme in the top PEG rich phase was then extracted into a Na₂HPO₄ rich bottom phase and further concentrated by ultrafiltration. The partitioning behavior of amyloglucosidase was examined in PEG/MDX, PEG/Na₂SO₄, PEG/Na₂HPO₄, PEG/KH₂PO₄ aqueous two phase systems. Effect of buffering salts such as NaCl, Na₂HPO₄, KH₂PO₄ and Na₂SO₄ on the partitioning behavior of enzyme was studied in PEG/MDX system. Maximum partitioning of amyloglucosidase was seen with KH₂PO₄ (m = 18.1). A two stage ATPE employing PEG/MDX (buffered with KH₂PO₄) and PEG/Na₂HPO₄ systems, followed by ultrafiltration has resulted in an overall recovery of 78.4% with 3.1 fold purification and 9.4 fold concentration of the enzyme.     

A proton magnetic relaxation study of human ferrihaemoglobin in aqueous salt solutions

The proton magnetic relaxation time, T₁, has been measured at 29 MH_z in 0.1M KH₂PO₄ and 0.1M NaCI (both pH 6) aqueous solutions of human ferrihaemoglobin, the protein concentrations ranging from 0.5 to 5 mM per haem. The linear dependence on protein concentration of the enhancement in relaxation rates, Δ(1/T₁), due to the presence of the paramagnetic iron in haemoglobin was confirmed at 34°C and at ～10°C. In the middle temperature range there is a thermally activated process, whose energy of activation depends on protein concentration. This dependence is different for the two salt solutions; E_a increases with c_Hb for 0.1M KH₂PO₄ and decreases for 0.1M NaCI. The model of water-proton exchange between the bulk solvent and the sixth coordination site of the haem iron was used to calculate the distance from the “liganded” water protons to the haem iron. This yields distances much larger than those determined by X-ray crystal structure analysis. A model is proposed which reconciliates both types of data. The low-temperature relaxation rates cannot be used in deriving quantitative stereochemical data for the haem pocket because of its special shape. Irrespective of the molecular model adopted, the experimental results show clearly that, both at low (～10°C) and higher (>34°C) temperatures, the interaction of paramagnetic haem iron with water protons is practically the same for the two aqueous solutions. The dynamic state of the haemoglobin molecule, as indicated by the middle-temperature range, is completely different in 0.1M KH₂PO₄ and 0.1M NaCl, pH 6.     

The effects of temperature and IAA concentration on the latent period for IAA-induced rapid growth of Avena coleoptile segments

Summary Indoleacetic acid buffered at pH 7.0 induces a high growth rate in Avena coleoptile segments after a latent period, the duration of which is dependent upon both IAA concentration and temperature. A minimum latent period of 7.3 min is observed at 25° C with 10^-3 M IAA in phosphate buffer at pH 7.0.In contrast, 5×10^-3 M IAA made up in 0.01 M KH₂PO₄ alone, promotes elongation almost immediately, regardless of whether the segments have been previously incubated in 0.01 M KH₂PO₄ at pH 4.7, or phosphate buffer at pH 7.0. This immediate response is unaffected by 10^-4 M KCN which abolishes the rapid growth induced by 5×10^-3 M IAA buffered at pH 7.0 but does not affect the immediate appearance of low-pH-induced growth. Since we consistently find solutions of 5×10^-3 M IAA in 0.01 M KH₂PO₄ to have a pH of 3.5, our results indicate that the immediate growth response elicited by this solution is attributable to its low pH rather than to the presence of IAA as previously reported in the literature.     

Histochemical demonstration of carbonic anhydrase activity

Summary Freeze-dried frozen sections are floated on the surface of the freshly prepared incubation mixture (CoSO₄ 1.75 × 10^–3 M, H₂SO₄ 5.3 × 10^–2 M, NaHCO₃ 1.57 × 10^–2 M and KH₂PO₄ 1.17 to 11.7 × 10^–3 M; demonstration of weak activity requires high phosphate). A compound containing cobalt and phosphorous precipitates at carbonic anhydrase sites and is converted to CoS. Adequate staining requires only 2–10 minutes of incubation. Actazolamide inhibits the staining reaction in specific concentrations. Actazolamidein vivo, 20 mg/kgi.v. to mice 30 minutes before sacrifice also inhibited the staining. The proportion phosphorous in the specific precipitate increases with KH₂PO₄ of the medium (shown by the addition of⁶⁰Co and³²P). An explanation of the reaction mechanism is given, based on the catalyzed loss of CO₂ in the surface layer. The inclusion of phosphate in the medium makes this modification ofHäusler's method so sensitive that it shows carbonic anhydrase activity in for instance stratum spinosum of the skin.This investigation was supported by grants from the Medical Faculty, University of Uppsala and from the U.S. National Institutes of Health (Grant NB 3060 to E.Bárány).     

Recovery of antibiotics by aqueous two-phase partition —Partitioning behavior of pure acetylspiramycin solution in polyethylene glycol/potassium phosphate aqueous two-phase systems

Summary The effects of average molecular weight of PEG, concentrations of PEG and KH₂PO₄ and pH on the partition equilibrium of acetylspiramycin in PEG/KH₂PO₄ aqueous two-phase systems were studied in detail. The partition coefficients of acetylspiramycin in PEG/ KH₂PO₄ systems were measured at room temperature 25 °C. It was found that acetylspiramycin partitioned unevenly in the aqueous two-phase systems composed of PEG and KH₂PO₄ and could be purified by this technique. A suitable phase-forming system (pH=6.7, 12w/w% PEG2000, 11w/w% KH₂PO₄) was found out after partition coefficient (Kp=42) , extraction ratio (=96%) and recovery ratio(R=98.8%) were investigated comprehensively in this paper.Hua qiang is one of the cooperators of the experimetal.     

Influence of the phosphorus and nitrogen source on nisin production in Lactococcus lactis subsp. lactis batch fermentations using a complex medium

The influence of different phosphorus and nitrogen sources on Lactococcus lactis subsp. lactis NIZO 22186 growth and nisin production was studied in batch fermentations using a complex medium. KH₂PO₄ was found to be the best phosphorus source for nisin production. Increasing initial phosphate concentrations from 0 to 5% KH₂PO₄ exerted a double effect, creating favourable pH conditions and particularly stimulating the nisin production levels, which were highest at 5% KH₂PO₄. Up to now, no such high initial phosphate concentrations have been reported for the production of other antibiotics or bacteriocins. Nisin, a lanthionine-containing peptide antibiotic with bacteriocin properties, clearly behaved as a primary metabolite, since its formation was linked with active growth and was not suppressed by phosphate concentrations up to 5%. A complex medium supplemented with cotton seed meal as nitrogen source also gave very high nisin yields. Correspondence to: L. De Vuyst     

Batchwise Purification of Liver Ribosomes and Polysomes from Unfasted Mice on Hydroxylapatite

Batchwise purification of liver ribosomes and polysomes on hydroxyl-apatite is a rapid procedure to remove glycogen, hemoglobin, ribonuclease and other contaminants from ribosomal preparations. Ribosomes and polysomes are adsorbed to hydroxylapatite in a Büchner filter funnel and the contaminants are eluted from the hydroxylapatite with 0.15 M KH₂PO₄. The ribosomes and polysomes are then eluted with 0.3–0.4 M KH₂PO₄ and concentrated by centrifugation. The resolution of the polysome profiles was greatly improved following purification. The purified ribosomes could be dissociated into subunits at 0.3 M KCl, and showed no loss of activity in poly-U directed phenylalanine synthesis.     

Increasing phosphorus concentration in the extraradical hyphae of <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> DAOM 197198 leads to a concomitant increase in metal minerals

Plants associated with arbuscular mycorrhizal fungi (AMF) acquire phosphorus via roots and extraradical hyphae. How soil P level affects P accumulation within hyphae and how P in hyphae influences the accumulation of metal minerals remains little explored. A bi-compartmented in vitro cultivation system separating a root compartment (RC), containing a Ri T-DNA transformed carrot root associated to the AMF Rhizophagus irregularis DAOM 197198, from a hyphal compartment (HC), containing only the extraradical hyphae, was used. The HC contained a liquid growth medium (i.e., the modified Strullu-Romand medium containing P in the form of KH₂PO₄) without (0 μM) or adjusted to 35, 100, and 700 μM of KH₂PO₄. The accumulation of P and metal minerals (Ca, Mg, K, Na, Fe, Cu, Mn) within extraradical hyphae and AMF-colonized roots, and the expression of the phosphate transporter gene GintPT were assessed. The expression of GintPT in the extraradical hyphae did not differ in absence of KH₂PO₄ or in presence of 35 and 100 μM KH₂PO₄ in the HC but was markedly reduced in presence of 700 μM KH₂PO₄. Hyphal P concentration was significantly lowest in absence of KH₂PO₄, intermediate at 35 and 100 μM KH₂PO₄ and significantly highest in presence of 700 μM KH₂PO₄ in the HC. The concentrations of K, Mg, and Na were positively associated with the concentration of P in the extraradical hyphae developing in the HC. Similarly, P concentration in extraradical hyphae in the HC was related to P concentration in the growth medium and influenced the concentration of K, Mg, and Na. The accumulation of the metal mineral K, Mg, and Na in the extraradical hyphae developing in the HC was possibly related to their function in neutralizing the negative charges of PolyP accumulated in the hyphae.     

A kinetic study of phosphorus absorption by excised maize roots from flowing solutions with different phosphorus concentrations

The effect of two different mechanisms of phosphorus ion transport from the nutrient solution volume to the surface areas of excised maize roots was studied under concentrations ranging from 0.01 mM to 50.0 mM KH₂PO₄. A modified technique of study of kinetic ion absorption was used. In the control series, the roots were placed in absorption solution without flow (the dominant mechanism of ion transport to the roots being diffusion), while in the experimental series the absorption solution was flowing round the roots at a rate of 0.162 cm s^?1 (the dominant mechanism of ion transport to the roots being mass flow). The rate of phosphorus absorption by the roots from flowing solutions was highly significantly increased at all concentrations of absorption solution except for the 50.0 mM KH₂PO₄ concentration. The increase in phosphorus absorption in the case of 50.0 mM KH₂PO₄ concentration was non-significant due to the fact that the high concentration of phosphorus together with the diffusion of phosphorus ions ensured a sufficient supply of phosphorus to the roots, covering the requirement for their uptake. The results point to the need for an analysis of environmental factors to be carried out in studying ion absorption kinetics, and reveal the inadequacy of methods usually employed in such investigations, in particular with respect to the homogeneity of the nutrient solution in the whole of its volume and especially round the roots.     

Tuberculate spore formation by thirty-two strains of Histoplasma capsulatum

Summary 1. Thirty-two strains ofHistoplasma capsulatum were studied concerning their ability to form tuberculate spores and their conversion into the yeast phase.2. Nine strains did not produce tuberculate spores on Sabouraud's agar, on corn meal agar, on spent medium, on media with pH adjusted from 4.5 to 7.0 or on the first passage through hamsters.3. Tuberculate spore production did occur in these nine strains when Sabouraud's medium was enriched with phosphate, especially KH₂PO₄. In addition, all but two strains produced tuberculate spores after a second passage through hamsters.4. Growth on KH₂PO₄ enriched Sabouraud's agar led to a greater yield of yeast phase as compared to yeast phase resulting from colonies of the same strain grown on plain Sabouraud's agar. This may be due to the greater number of spores produced on the KH₂PO₄ enriched medium.5. A grinding technique of preparing inocula improved slightly the facility of obtaining the yeast phase over heavy inoculation with unground pieces of mold culture.