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1.
The availability of six organic phosphorus substrates (OPS)to serve as sources of this nutrient for natural populationsof phytoplanktonic algae and bacteria was tested by measuringthe degree to which the organic substrates decreased uptakeof [32P]orthophosphate (32Pi). When added to samples of LakeKinneret water, six organo-phosphorus compounds usually loweredthe amount of 32Pi] taken up by microplankton retained on 0.2-µfilters.In contrast, OPS addition generally stimulated 32Pi uptake intothe mainly algal fraction (>3 µm), indicating thatthe bacteria were mostly responsible for utilising OPS. Thesparing effect of OPS addition on 32P1 uptake was very fast,suggestingthat the micro-organisms possessed constitutive or rapidly inducedenzyme systems to exploit the OPS.The results of this studyindicate that a significant flux of phosphorus may pass viaDOP into microbiota, especially bacteria, in some aquatic systems.  相似文献   

2.
Skeletal muscle fiber types differ in their contents of total phosphate, which includes inorganic phosphate (Pi) and high-energy organic pools of ATP and phosphocreatine (PCr). At steady state, uptake of Pi into the cell must equal the rate of efflux, which is expected to be a function of intracellular Pi concentration. We measured 32P-labeled Pi uptake rates in different muscle fiber types to determine whether they are proportional to cellular Pi content. Pi uptake rates in isolated, perfused rat hindlimb muscles were linear over time and highest in soleus (2.42 ± 0.17 µmol·g–1·h–1), lower in red gastrocnemius (1.31 ± 0.11 µmol·g–1·h–1), and lowest in white gastrocnemius (0.49 ± 0.06 µmol·g–1·h–1). Reasonably similar rates were obtained in vivo. Pi uptake rates at plasma Pi concentrations of 0.3–1.7 mM confirm that the Pi uptake process is nearly saturated at normal plasma Pi levels. Pi uptake rate correlated with cellular Pi content (r = 0.99) but varied inversely with total phosphate content. Sodium-phosphate cotransporter (PiT-1) protein expression in soleus and red gastrocnemius were similar to each other and seven- to eightfold greater than PiT-1 expression in white gastrocnemius. That the PiT-1 expression pattern did not match the pattern of Pi uptake across fiber types implies that other factors are involved in regulating Pi uptake in skeletal muscle. Furthermore, fractional turnover of the cellular Pi pool (0.67, 0.57, and 0.33 h–1 in soleus, red gastrocnemius, and white gastrocnemius, respectively) varies among fiber types, indicating differential management of intracellular Pi, likely due to differences in resistance to Pi efflux from the fiber. inorganic phosphate; sodium-inorganic phosphate transporters; PiT-2; inorganic phosphate efflux  相似文献   

3.
We studied the kinetics of inorganic phosphate (P1) uptake from0.1–1,000 µM P1 by protoplasts from suspension-culturedcells of Catharanthus roseus (L.) G. Don. Concentration dependenceof [32P]P1 uptake revealed two kinetically different uptakesystems, a high-affinity system and a low-affinity system, withKm values of 3.0 and 47 µM, respectively. Protoplastsfrom cells grown in Pi-rich media had a medium level of thelow-affinity activity and a very low level of the high-affinityactivity. It appeared low-affinity system is expressed constitutively,while the high-affinity system is regulated by the availabilityof Pi. When cells grown in a Pi-rich media were transferredto Pi-depleted media, the high-affinity activity increased significantlyafter 2 d, but the low-affinity activity was barely changed.Upon addition of 10 mM Pi, the high level of the high-affinityactivity fell to almost undetectable level in 1d. Both uptakesystems exhibited maximum activity between pH 5 and 6. 1 Present address: Tokyo Research Laboratories, Kyowa HakkoKogyo Co., Ltd., 3-6-6 Asahi-cho, Machida, Tokyo, 194 Japan.  相似文献   

4.
To study possible changes in the transport metabolites betweenchloroplasts and cytoplasm during CAM induction of Mesembryanthemumcrystallinum, we compared substrate specificity of P11 translocator(s)in isolated chloroplasts from the C3 and CAM-induced plants.The [14C]glu-cose 6-phosphate (G6P) transport activity was significantonly in the chloroplasts of CAM-mode plants and not detectablein those of C3-mode, while a similar high rate of [32P]Pi uptakewas observed with both types of chloroplasts. Kinetic analysisof G6P uptake in the CAM chloroplasts showed a high Vmax [10.6µmol (mg Chl)–1 h–1] and a comparatively lowKm value (0.41 mM); the latter was similar to Ki values of Pi,3-phosphoglycerate and phospho-enolpyruvate, 0.30, 0.34 and0.47 mM, respectively. On the other hand, [32P]Pi uptake inthe CAM chloroplasts was inhibited competitively by G6P witha Ki value (8.4 mM) 20-fold higher than the Km value for G6Puptake, while that in C3 chloroplasts was not inhibited at all.These results suggest that a new G6P/Pi, counterexchange mechanismis induced in the chloroplast envelope of CAM-induced M. crystallinumin addition to the ordinary type of P, translocator, that cannottransport G6P, already present in the C3-type chloroplasts. (Received March 17, 1997; Accepted May 10, 1997)  相似文献   

5.
The isotope 15N was used to examine nitrogen dynamics in LakesFryxell and Vanda, two permanently ice-covered Antarctic lakes.Half-saturation constants for NH4+. uptake in the shallow watersof both lakes were <10 µg N l–1; uptake kineticexperiments on populations forming the deep-chlorophyll layersof these lakes showed zero-order kinetics and could not be fittedwith the Michaelis-Menten equation. Elevated uptake within thefirst few minutes following pulses of NH4+. and NO3 occurredin both lakes. NH4+ regeneration, determined from isotope dilutionexperiments, exceeded uptake at 4.6 m in Lake Fryxell, was lessthan uptake at 9 m in Lake Fryxell and was equal to uptake at10 m in Lake Vanda under the experimental conditions. NO3uptake was suppressed by NH4+ levels as low as 2 µg NH4+-N l–1 in Lake Fryxell; the suppression was strongestin the near-surface populations. Substrate-saturated C:N uptakeratios (g:g) in Lake Fryxell decreased from 8.4 near the surfaceto 1.8 at the bottom of the trophogenic zone. Overall, the nitrogendynamics in these lakes are similar to other lakes and the openocean in that biological productivity during the austral summeris supported by regenerated nutrients.  相似文献   

6.
The general phosphate need in mammalian cells is accommodated by members of the Pi transport (PiT) family (SLC20), which use either Na+ or H+ to mediate inorganic phosphate (Pi) symport. The mammalian PiT paralogs PiT1 and PiT2 are Na+-dependent Pi (NaPi) transporters and are exploited by a group of retroviruses for cell entry. Human PiT1 and PiT2 were characterized by expression in Xenopus laevis oocytes with 32Pi as a traceable Pi source. For PiT1, the Michaelis-Menten constant for Pi was determined as 322.5 ± 124.5 µM. PiT2 was analyzed for the first time and showed positive cooperativity in Pi uptake with a half-maximal activity constant for Pi of 163.5 ± 39.8 µM. PiT1- and PiT2-mediated Na+-dependent Pi uptake functions were not significantly affected by acidic and alkaline pH and displayed similar Na+ dependency patterns. However, only PiT2 was capable of Na+-independent Pi transport at acidic pH. Study of the impact of divalent cations Ca2+ and Mg2+ revealed that Ca2+ was important, but not critical, for NaPi transport function of PiT proteins. To gain insight into the NaPi cotransport function, we analyzed PiT2 and a PiT2 Pi transport knockout mutant using 22Na+ as a traceable Na+ source. Na+ was transported by PiT2 even without Pi in the uptake medium and also when Pi transport function was knocked out. This is the first time decoupling of Pi from Na+ transport has been demonstrated for a PiT family member. Moreover, the results imply that putative transmembrane amino acids E55 and E575 are responsible for linking Pi import to Na+ transport in PiT2. inorganic phosphate transport; retroviral receptor; SLC20  相似文献   

7.
Vitamins, phytoplankton and bacteria: symbiosis or scavenging?   总被引:1,自引:0,他引:1  
The conclusion that over 25% of global primary production dependson direct algal/bacterial symbiosis involving vitamin B12 [Croftet al., (2005) Algae acquire vitamin B12 through a symbioticrelationship with bacteria. Nature, 438, 90–93] is patentlyfalse, for it is based on a misconception of the probable levelof the vitamin B12 requirement in marine pelagic algae. A reviewof the various published attempts at measuring this requirementsuggests that it is likely to be so low that oceanic and coastalconcentrations of the vitamin would usually be sufficient tosustain the populations that occur without the assistance ofdirect algal/bacterial symbiosis. The levels measured are discussedin relation to method (batch or continuous culture) and protocolsused. Requirement values considered by the author to be acceptablerange from 0.1 to 0.3 pM for the vitamin growth saturation constant(KS) and from 30 to 100 µL algal biomass pmol–1vitamin for the yield.  相似文献   

8.
Autotrophic picoplankton populations in Lake Kinneret are composedof picocyanobacteria and picoeukaryotes. Overall, the ratesof photosynthetic carbon fixed by autotrophic picoplankton duringthis study were low (0.01–1.5 mg Cm–3 h–1).The highest chlorophyll photosynthetic activity of the <3µm cell-size fraction was found in spring, when picoeukaryotespredominated and in addition small nanoplankton passed throughthe filters. The maximum cell-specific photosynthetic rate ofcarbon fixation by picocyanobacteria and picoeukaryotes was2.5 and 63 fg C cell–1 h–1, respectively. The highestspecific carbon fixation rate of autotrophic picoplankton was11 µg C µg–1 Chl h–1 The proportionalcontribution of autotrophic picoplankton to total photosynthesisusually increased with depth. Picocyanobacteria collected fromthe dark, anaerobic hypolimnion were viable and capable of activephotosynthesis when incubated at water depths within the euphoticzone. Maximum rates of photosynthesis (Pmax) for picocyanobacteriaranged from 5.4 to 31.4 fg C cell–1 h–1 with thehighest values in hypolimnetic samples exposed to irradiance.Photosynthetic efficiency (  相似文献   

9.
Small, aloricate ciliates dominated the biomass of heterotrophicprotists throughout the water column at the end of the periodof stratification in Lake Kinneret, Israel The integrated biomassof cilates was 5–20 times that of heterotrophic flagellatesDuring incubation experiments, ciliate growth rates in cpilimneticwater corresponded to population doubling times of 9.6–19.4h, while flagellate populations showed no growth. Most of thealiates were small forms (10–30 µm long), includingscuticocihates, choreotnchs, Coleps spp. and Colpoda spp., andappeared to be consuming bacteria, coccoid cyanobacteria, and<5 µm eukaryotic algae. Grazing rates of cihate assemblageson picoplankton in the epilimnion, as determined by the uptakeof fluorescently labeled bacteria and cyanobactena, ranged from62 to 86 nl cell1 h1 Colpoda steini, isolatedfrom lakewater, grew on a cultured freshwater Synechococcussp with a doubling time of 4.5 h, and a gross growth efficiencyof 48% The estimated daily requirements of ciliates for growthapproximately equalled total phytoplankton production. We calculatedthat ciliates in the epilimnion were clearing 4–10% ofthe bacterioplankton and cyanobactenal standing stocks per daySince this would not be sufficient food consumption to meetdaily carbon requirements of the aliates, it is likely thatthese organisms were also grazing a significant amount of autotrophicand heterotrophic eukaryotic cells in Lake Kinneret.  相似文献   

10.
A search was made for inhibitors of Pi uptake that act directlyon the Pi transporter in the plasma membranes of Catharanthusroseus cells to inhibit Pi uptake without inhibition of protonpumping. Using standard electrodes, we monitored changes inpH and in the concentration of K+ ions, as well as the rateof Pi uptake, when an inhibitor to be tested was applied tothe cells in unbuffered medium. A9C (28 µM), a blockerof anion channels, inhibited Pi uptake but it also inhibitedthe proton pump. However, a structurally similar inhibitor,furosemide, inhibited Pi uptake without inhibiting proton pumping. It is suggested that the carboxylic group of these inhibitorsinteracts with the Pi-binding site (probably an amino group)of the Pi transporter in the plasma membrane and that the hydrophobicstructure of these inhibitors facilitates their accumulationin the plasma membrane. 3Present address: Department of Biology, Hitotsubashi University,2-1 Naka, Kunitachi, Tokyo, 186 Japan  相似文献   

11.
The influence of algal cell size on the light environment withindense phytoplankton populations was analysed in cultures of11 freshwater algae (mostly Chlorophyta) varying widely in cellsize and growing at saturated (220 µM m2 s1) andlimited (11 µM m2 s1) irradiances Chlorophyll-a-specificabsorbance coefficients did not bear any obvious relationshipto cell size and, as expected from previous analyses, were stronglyinfluenced by the light intensity at which the algae were grown.However, biomass (fresh weight)-specific absorbance coefficientsdecreased as algal cell volume increased regardless of the incomingirradiance Intracellular chlorophyll a concentration (c1), whichdecreased as algal cell size increased, explained a substantialfraction of the variance in biomass (fresh weight)-specifjcabsorbance coefficients. Thus, this study supports the notionthat reduced serf-shading allows large algae to support greatermaximal biomass than smaller algae.  相似文献   

12.
The effect of adding dissolved substrates derived from algalcells on the patterns of nutrient cycling and growth of bacteria,heterotrophic nanoflagellates (HNAN) and photoautotrophs wasdetermined in samples of near-surface waters from Lake Kinneret.Supplementation of substrates always resulted in an increasedpeak of HNAN numbers and had little effect on bacterial numbers.HNAN-mediated nutrient remineralization of nitrogen and phosphoruswas also stimulated. In light-incubated samples the remineralizednutrients were taken up by photoautotrophic cells. Maximum growthrates observed for HNAN ranged from 0.03 to 0.11 h–1,clearance rates for bacteria 1.1–7.3 nl HNAN–1 h–1and remineralization rates 6.4–8.4 µg N mg dry wt–1h–1 and 0.37–0.99 µg P mg dry wt–1 h–1.  相似文献   

13.
Permeability coefficients (PS values) for CO2 of the plasmamembrane (PM) of the unicellular green algae Eremosphaera viridis,Dunaliella parva, and Dunaliella acidophila, and of mesophyllprotoplasts isolated from Valerianella locusta were determinedfrom 14CO2 uptake experiments using the rapid separation ofcells by the silicone oil layer centrifugation technique. Theexperimental PS values were compared with calculated numbersobtained by interpolation of Collander plots, which are basedon lipid solubility and molecular size, for D. parva cells,mesophyll protoplasts isolated from Spinacia oleracea, mesophyllcells and guard cells of Valerianella, and guard cell protoplastsisolated from Vicia faba. The conductivity of algal plasma membranes for CO2 varies between0.1 and 9 ? 10–6 m s–1, whereas for the plasmalemmaof cells and protoplasts isolated from leaves of higher plantsvalues between 0.3 and 11 ? 10–6 m s–1 were measured.By assuming that these measurements are representative for plantsand algae in general, it is concluded that the CO2 conductivityof algal PM is of the same order of magnitude as that of thehigher plant cell PM. Ps values of plasma membranes for CO2are lower than those for SO2, but are in the same order of magnitudeas those measured for H2O. On the basis of these results itis concluded that theoretical values of about 3000 ? 10–6m s–1 believed to be representative for higher plant cells(Nobel, 1983) and which are frequently used for computer-basedmodels of photosynthesis, lack experimental confirmation andrepresent considerable overestimations. However, with severalsystems, including higher plant cells, the conductance of thePM for CO2 was significantly higher in light than in darkness.This suggests that in light, additional mechanisms for CO2 uptakesuch as facilitated diffusion or active uptake may operate inparallel with diffusional uptake. Key words: Conductivity, CO2, permeability coefficient, photosynthesis, plasmalemma  相似文献   

14.
Members of the SLC20 family or type III Na+-coupled Pi cotransporters (PiT-1, PiT-2) are ubiquitously expressed in mammalian tissue and are thought to perform a housekeeping function for intracellular Pi homeostasis. Previous studies have shown that PiT-1 and PiT-2 mediate electrogenic Pi cotransport when expressed in Xenopus oocytes, but only limited kinetic characterizations were made. To address this shortcoming, we performed a detailed analysis of SLC20 transport function. Three SLC20 clones (Xenopus PiT-1, human PiT-1, and human PiT-2) were expressed in Xenopus oocytes. Each clone gave robust Na+-dependent 32Pi uptake, but only Xenopus PiT-1 showed sufficient activity for complete kinetic characterization by using two-electrode voltage clamp and radionuclide uptake. Transport activity was also documented with Li+ substituted for Na+. The dependence of the Pi-induced current on Pi concentration was Michaelian, and the dependence on Na+ concentration indicated weak cooperativity. The dependence on external pH was unique: the apparent Pi affinity constant showed a minimum in the pH range 6.2–6.8 of 0.05 mM and increased to 0.2 mM at pH 5.0 and pH 8.0. Xenopus PiT-1 stoichiometry was determined by dual 22Na-32Pi uptake and suggested a 2:1 Na+:Pi stoichiometry. A correlation of 32Pi uptake and net charge movement indicated one charge translocation per Pi. Changes in oocyte surface pH were consistent with transport of monovalent Pi. On the basis of the kinetics of substrate interdependence, we propose an ordered binding scheme of Na+:H2PO4:Na+. Significantly, in contrast to type II Na+-Pi cotransporters, the transport inhibitor phosphonoformic acid did not inhibit PiT-1 or PiT-2 activity. Na+-Pi cotransport; two-electrode voltage clamp; surface pH electrode; SLC20; retroviral receptor  相似文献   

15.
The relationship between chlorophyll a (Chl a) and primary productivity(PP) in the uppermost water layer and the water column-based(0–15 m) integral values of those variables were examinedusing measurements taken in Lake Kinneret (Israel) from 1990to 2003. In 81% of all Chl a profiles examined, the distributionwas fairly uniform within the entire 0–15 m water column,and 12.3% of instances showed a prominent subsurface maximum,when the lake phytoplankton was dominated by the dinoflagellatePeridinium gatunense. Chl a can be reliably estimated by remotesensing techniques in the productive and turbid water of LakeKinneret, since Chl a concentration at surface layers can beextrapolated to the entire water column. Light vertical attenuationcoefficient average for wavelengths from 400 to 700 nm, Kd,ranged from 0.203 to 1.954 m–1 and showed high degreeof temporal variation. The maximal rate of photosynthetic efficiency,PBopt [average 3.16 (±1.50)], ranged from 0.25 to 8.85mg C m–3 h–1 mg Chl a–1. Using measured dataof Chl a, PBopt, and light as an input, a simple depth-integratedPP model allowed plausible simulation of PP. However, a lackof correlation between photosynthetic activity and temperature(or other variable with remotely sensed potential) renders theuse of models that require input of photosynthetic efficiencyto calculate integrated PP of little value in the case of productiveand turbid Lake Kinneret.  相似文献   

16.
Rapid physiological assays for nutrient demand by the plankton. I. Nitrogen   总被引:3,自引:0,他引:3  
Three assays for nitrogen demand were compared on samples ofnatural plankton and on green and blue-green algal species inculture. The most reliable guide was selective luxury uptakeof nitrogen by the plankton after enrichment with a 10-to-1(wt/wt) mixture of inorganic N and P. Ammonium transport capacity,measured either directly (cultures) or by Vmax for uptake ofthe NH4+ analogue methylammonium (lakewaters), was generallyhigh in lakewaters with low dissolved inorganic N-to-P ratiosand in N-deficient cultures of green algae. By contrast, transportcapacity was much reduced in both natural and cultured populationsof heterocystous blue-green algae growing under conditions oflow combined inorganic nitrogen. Ammonium enhancement assays(heterotrophic CO2, fixation after NH4+ enrichment) were conductedat monthly intervals on eutrophic Lake Rotorua. There was astrong, negative correlation between this indicator of N deficiencyand dissolved inorganic N-to-P ratios below a threshold of 6:1.Ammonium enhancement was not, however, a reliable indicatorof combined inorganic N-demand by populations of heterocystousblue-green algae. All three assays provided strong evidenceof a persistent shortage of nitrogen relative to phosphorusfor algal production in Lake Rotorua.  相似文献   

17.
Mitochondria show extensive movement along neuronal processes, but the mechanisms and function of this movement are not clearly understood. We have used high-resolution confocal microscopy to simultaneously monitor movement of mitochondria and changes in intracellular [Ca2+] ([Ca2+]i) in rat cortical neurons. A significant percentage (27%) of the total mitochondria in cortical neuronal processes showed movement over distances of >2 µM. The average velocity was 0.52 µm/s. The velocity, direction, and pattern of mitochondrial movement were not affected by transient increases in [Ca2+]i associated with spontaneous firing of action potentials. Stimulation of Ca2+ transients with forskolin (10 µM) or bicuculline (10 µM), or sustained elevations of [Ca2+]i evoked by glutamate (10 µM) also had no effect on mitochondrial transit. Neither removal of extracellular Ca2+, depletion of intracellular Ca2+ stores with thapsigargin, or inhibition of synaptic activity with TTX (1 µM) or a cocktail of CNQX (10 µM) and MK801 (10 µM) affected mitochondrial movement. These results indicate that movement of mitochondria along processes is a fundamental activity in neurons that occurs independently of physiological changes in [Ca2+]i associated with action potential firing, synaptic activity, or release of Ca2+ from intracellular stores. calcium transient; dendrites  相似文献   

18.
Microbial potential uptake and regeneration rates of ammonium(NH4+) were studied along a salinity gradient (salinities 0.2–34.4)in the Mississippi River plume during an extreme drought inspring 2000. Chlorophyll concentrations up to 30 µg L–1were highest in the low- and mid-salinity regions (salinities8.5–28.2) and comparable to records of other years butextended over smaller areas than during periods of normal riverflow. Bacterial biomass (5.1–28.3 µg C L–1)was at the low end of the range observed in normal flow years,decreased with distance from the river mouth and did not peakwith chlorophyll. Heterotrophic nanoflagellate abundance (1.4–4.0µg C L–1) did not reflect phytoplankton and bacterialspatial distribution but peaked at 9.2 µg C L–1at salinity 8.5. Microbial NH4+ regeneration rates were estimatedby 15NH4+ isotope dilution experiments for the whole microbialcommunity, under light and dark conditions, and for the <2µm bacterium-dominated size fraction. Microbial NH4+ regenerationrates (0.018–0.124 µmol N L–1 h–1) werelow relative to previous reports and peaked at salinity 28.Total NH4+ regeneration rates were higher than those in the<2 µm size fraction at only four stations, suggestingthat bacterial mineralization was a significant component ofNH4+ recycling in some parts of the river plume. Higher NH4+regeneration in whole-water samples versus <2 µm fractionsprovided evidence for microbial grazing in regions where chlorophylland regeneration rates peaked and at two full-salinity stations.  相似文献   

19.
We examined the NO3 and NH4+ uptake capabilities of thedinoflagellate Peridinium cinctum and of the accompanying nanoplanktonduring the spring P. cinctum bloom in Lake Kin-neret. Throughoutthe Peridinium season, the smaller algae had greater affinitiesand faster specific uptake rates for both NO3and NH4+It appears that P. cinctum cannot directly compete with nanoplanktonfor nitrogen nutrients. Other factors such as the ability ofdinoflagellates to swim freely in the water column and low grazingpressures may explain their dominance in the lake.  相似文献   

20.
A strain of Coleps hirtus (Ciliophora, Prorodontida) was isolatedfrom the epilimnion of monomictic Lake Kinneret. Growth of thisciliate was tested in response to 12 species of planktonic algaeand seven species of cultured bacteria from lake isolates whichwere offered as food. Eight species of algae (one Cryptophyceaeand seven Chlorophyceae) and four bacteria supported good toexcellent growth of C.hirtus. Growth rates (µ) and doublingtimes (DT) ranged from 0.008 to 0.029 h–1 and from 23.9to 90.8 h respectively. C.hirtus was able to grow on bacteriaat concentration levels as low as 2–8 x 105 cells ml–1.No correlation was observed between growth rate of C.hirtusand cell volume of the prey. aPresent address: Istituto di Ecologia, Universita di Parma,43100 Parma, Italy  相似文献   

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