Transfer Of 32P Research Articles

Defence responses of native and invasive plants to the native generalist vine parasite Cassytha pubescens – anatomical and functional studies

We investigated the responses of two invasive and two native host species to the parasitic vine Cassytha pubescens R.Br. using glasshouse experiments. We assessed growth of the parasite and its hosts, and anatomy and functionality of haustoria. Target hosts were infected using C. pubescens already established on a donor host. This enabled measurement of growth in target hosts that were detached (parasite connection severed) or not from the donor host. Haustorial connections to hosts were investigated using histological methods. We tested the functionality of haustoria in one invasive and one native host using radiolabelled phosphorus (32P). After it was severed from the donor host, C. pubescens grew poorly on the native host, Acacia myrtifolia (Sm.)Willd. This was likely due to a lack of effective functional haustorial development because although haustoria were firmly attached and morphologically alike those formed on the other hosts, their anatomy was different: their connections with the vascular system were not developed and there was no transfer of 32P from A. myrtifolia to the parasite. In contrast, the other three host species supported the growth of the parasite and had fully developed haustoria. Effective transfer of 32P from the invasive host to the parasite confirmed this. Our results suggest a range of defence mechanisms in C. pubescens hosts and are consistent with reports of strong detrimental effects on invasive hosts. Further, they amount to evidence for the potential use of a native parasite as biological control for invasive species.

Incorporation of phosphate into glycogen by glycogen synthase

The storage polymer glycogen normally contains small amounts of covalently attached phosphate as phosphomonoesters at C2, C3 and C6 atoms of glucose residues. In the absence of the laforin phosphatase, as in the rare childhood epilepsy Lafora disease, the phosphorylation level is elevated and is associated with abnormal glycogen structure that contributes to the pathology. Laforin therefore likely functions in vivo as a glycogen phosphatase. The mechanism of glycogen phosphorylation is less well-understood. We have reported that glycogen synthase incorporates phosphate into glycogen via a rare side reaction in which glucose-phosphate rather than glucose is transferred to a growing polyglucose chain (Tagliabracci et al. (2011) Cell Metab13, 274–282). We proposed a mechanism to account for phosphorylation at C2 and possibly at C3. Our results have since been challenged (Nitschke et al. (2013) Cell Metab17, 756–767). Here we extend the evidence supporting our conclusion, validating the assay used for the detection of glycogen phosphorylation, measurement of the transfer of 32P from [β-32P]UDP-glucose to glycogen by glycogen synthase. The 32P associated with the glycogen fraction was stable to ethanol precipitation, SDS-PAGE and gel filtration on Sephadex G50. The 32P-signal was not affected by inclusion of excess unlabeled UDP before analysis or by treatment with a UDPase, arguing against the signal being due to contaminating [β-32P]UDP generated in the reaction. Furthermore, [32P]UDP did not bind non-covalently to glycogen. The 32P associated with glycogen was released by laforin treatment, suggesting that it was present as a phosphomonoester. The conclusion is that glycogen synthase can mediate the introduction of phosphate into glycogen, thereby providing a possible mechanism for C2, and perhaps C3, phosphorylation.

Native Grass Facilitates Mycorrhizal Colonisation and P Uptake of Tree Seedlings in Two Anthropogenic Substrates

Two microcosm experiments were conducted to study the role of extraradical mycelium (ERM) of arbuscular mycorrhizal fungi (AMF) in establishment and growth of tree species used for revegetation in anthropogenic substrates. Inoculated or non-inoculated Acer pseudoplatanus, Alnus glutinosa or Salix purpurea seedlings were grown with Calamagrostis epigejos (a grass spontaneously colonising degraded ecosystems) in two substrates (fly ash and coal mine spoil) either in direct root contact or in rhizoboxes with interaction only via ERM network. In both experiments, inoculation with AMF mostly had a positive effect on the growth of trees and increased the aggregation of fly ash. When plants grew in direct root-to-root contact, grass presence negatively affected tree growth, but it significantly improved mycorrhizal development (colonization of tree seedlings, spore number and ERM length). When grass and tree roots interacted via the ERM network, tree seedlings were successfully colonised by the ERM spreading from the C. epigejos roots. Mechanical disturbance of the ERM links between plants reduced AMF development and tree height in both substrates, but tree shoot biomass was not affected. In fly ash, inoculated, non-disturbed treatments showed significant transfer of 32P from the grass to the tree seedlings. It can be concluded that roots of A. pseudoplatanus, A. glutinosa or S. purpurea seedlings can be colonised from the ERM network radiating from quickly growing grasses, which can act as important agents for AMF distribution and facilitate mycorrhization of planted trees. In particular for willow, grass seems to be an essential nurse plant to achieve successful root colonisation.

Mouse Sir2 Homolog SIRT6 Is a Nuclear ADP-ribosyltransferase

Members of the Sir2 family of NAD-dependent protein deacetylases regulate diverse cellular processes including aging, gene silencing, and cellular differentiation. Here, we report that the distant mammalian Sir2 homolog SIRT6 is a broadly expressed, predominantly nuclear protein. Northern analysis of embryonic samples and multiple adult tissues revealed mouse SIRT6 (mSIRT6) mRNA peaks at day E11, persisting into adulthood in all eight tissues examined. At the protein level, mSIRT6 was readily detectable in the same eight tissue types, with the highest levels in muscle, brain, and heart. Subcellular localization studies using both C- and N-terminal green fluorescent protein fusion proteins showed mSIRT6-green fluorescent protein to be a predominantly nuclear protein. Indirect immunofluorescence using antibodies to two different mSIRT6 epitopes confirmed that endogenous mSIRT6 is also largely nuclear. Consistent with previous findings, we did not observe any NAD+-dependent protein deacetylase activity in preparations of mSIRT6. However, purified recombinant mSIRT6 did catalyze the robust transfer of radiolabel from [32P]NAD to mSIRT6. Two highly conserved residues within the catalytic core of the protein were required for this reaction. This reaction is most likely mono-ADP-ribosylation because only the modified form of the protein was recognized by an antibody specific to mono-ADP-ribose. Surprisingly, we observed that the catalytic mechanism of this reaction is intra-molecular, with individual molecules of mSIRT6 directing their own modification. These results provide the first characterization of a Sir2 protein from phylogenetic class IV.

Signal transduction abnormalities in melancholic depression

Intracellular signal transduction cascades, particularly those linked to protein kinases A (PKA) and C (PKC), have been implicated in mood disorders. This study examined the activity of PKA and PKC, as well as levels of PKA regulatory (R) and catalytic (C) subunit proteins, in fibroblasts cultured from skin biopsies from patients with major depression, melancholic subtype, in contrast to non-melancholic depressives and controls (n = 12 each group). PKA activity was determined as a function of the transfer of 32P to a target polypeptide, Kemptide. R and C subunit expression was assayed in the melancholic depressed and normal control groups by Western blots. In a separate experiment, the degree of phosphorylation of the endogenous substrate cAMP response element-binding protein (CREB) was estimated in samples from melancholic and non-melancholic patients and normal controls (n = 8 each) after incubation with isoproterenol or phorbol ester, which activate PKA and PKC respectively. Melancholics had significantly reduced phosphorylation of Kemptide in contrast to non-melancholics and controls. This was associated with lower levels of PKA RII alpha, C alpha, and C beta subunit isoform proteins, but not RI alpha, RI beta, or RII beta. Furthermore, activation of both PKA and PKC was associated with reduced CREB-P in melancholics relative to normal controls. Finally, PKA activity was found to correlate positively with Hamilton depression scores after 16 weeks of treatment with serotonin reuptake inhibitor antidepressants. These data further implicate signal transduction abnormalities in melancholic major depression, particularly PKA and PKC. This suggests an abnormality of factors controlling the expression or degradation of these enzymes.

Prostacyclin production in rat aortic smooth muscle cells: role of protein kinase C, phospholipase D and cyclooxygenase-2 expression

The present study was designed to investigate the role of protein kinase C (PKC) and phospholipase D (PLD) in angiotensin II (AngII)- and phorbol ester (PMA)-induced cyclooxygenase-2 (COX-2) expression and prostacyclin (PGI(2)) production in rat aortic smooth muscle cells (VSMC). Prostacyclin production in cultured VSMC was determined by radioimmunoassay. PKC activity was examined by measuring the transfer of 32P from (gamma-32P)ATP to histone III-S. COX-2 expression was determined by Western blotting. To measure PLD activity, thin layer chromatography was used. AngII (50 nM) and PMA (100 nM) promoted the translocation of PKC activity from the cytosol to the membranes within 30 min, followed by a strong increase in PLD activity as well as COX-2 expression and PGI(2) production. After 48 h exposure to PMA, PKC was downregulated resulting in a complete suppression of its activity. PKC-downregulation and the PKC inhibitor CGP41251 abolished PMA- and AngII-induced PLD activation, suppressed the stimulatory effect of PMA on COX-2 expression and PGI(2) production and strongly inhibited that of AngII. Furthermore, AngII- and PMA-induced PGI(2) production depended on protein synthesis and COX-2 but not COX-1 activity. Inhibition of PLD-mediated phosphatidic acid (PA) formation by 1% 1-butanol abolished AngII-induced COX-2 expression and PGI(2) secretion, while dioctanoyl PA increased COX-2 expression and PGI(2) production in a time- and concentration-dependent manner. Our results indicate that in VSMC, AngII promotes PGI(2) production to a large extent through a rise in COX-2 expression which is mediated by PA generated from increased PKC-dependent PLD activity.

Evaluation of the biological transfer of 32P, 137Cs and 65Zn by fish in the Yenisei River

The biological transfer of three radionuclides (32P, 137Cs, 65Zn) by fish in the Yenisei River (Central Siberia, Russia) was evaluated using a radioecological model. The modelling is based on the general ECOMOD methodology, where radionuclide behavior in an aquatic organism is linked with the processes of growth and metabolism, also with the concentrations of stable analogous elements in the organism, its food and the environment. The model was applied to explain the peculiarities of 32P, 137Cs and 65Zn accumulation in different ecological groups of fish, including non-migratory and migratory fish, non-predatory and predatory fish species. The highest activity concentrations in non-migratory fish from the Yenisei River were found for 32P. The accumulation of 32P by fish was shown to depend on the fish size (age, weight); however, it did not depend on the trophic status of fish. The modelling approach was developed to evaluate the biological transfer of radionuclides by the migratory fish, which spend the most part of life in the Yenisei delta, inlet or bay, and go upstream the Yenisei River for spawning. The results of the ECOMOD model calculations are in good agreement with available measurement data.

Evaluation of the biological transfer of 32P, 137Cs and 65Zn by fish in the Yenisei River

The biological transfer of three radionuclides ( 32P, 137Cs, 65Zn) by fish in the Yenisei River (Central Siberia, Russia) was evaluated using a radioecological model. The modelling is based on the general ECOMOD methodology, where radionuclide behavior in an aquatic organism is linked with the processes of growth and metabolism, also with the concentrations of stable analogous elements in the organism, its food and the environment. The model was applied to explain the peculiarities of 32P, 137Cs and 65Zn accumulation in different ecological groups of fish, including non-migratory and migratory fish, non-predatory and predatory fish species. The highest activity concentrations in non-migratory fish from the Yenisei River were found for 32P. The accumulation of 32P by fish was shown to depend on the fish size (age, weight); however, it did not depend on the trophic status of fish. The modelling approach was developed to evaluate the biological transfer of radionuclides by the migratory fish, which spend the most part of life in the Yenisei delta, inlet or bay, and go upstream the Yenisei River for spawning. The results of the ECOMOD model calculations are in good agreement with available measurement data.

Investigation of the kinetics and order of tyrosine phosphorylation in the T-cell receptor zeta chain by the protein tyrosine kinase Lck.

We report experiments to investigate the role of the physiologically relevant protein tyrosine kinase Lck in the ordered phosphorylation of the T-cell receptor zeta chain. Six synthetic peptides were designed based on the sequences of the immunoreceptor tyrosine-based activation motifs (ITAMs) of the zeta chain. Preliminary 1H-NMR studies of recombinant zeta chain suggested that it is essentially unstructured and therefore that peptide mimics would serve as useful models for investigating individual ITAM tyrosines. Phosphorylation kinetics were determined for each tyrosine by assaying the transfer of 32P by recombinant Lck on to each of the peptides. The rates of phosphorylation were found to depend on the location of the tyrosine, leading to the proposal that Lck phosphorylates the six zeta chain ITAM tyrosines in the order 1N (first) > 3N > 3C > 2N > 1C > 2C (last) as a result of differences in the amino-acid sequence surrounding each tyrosine. This proposal was then tested on cytosolic, recombinant T-cell receptor zeta chain. After in vitro phosphorylation by Lck, the partially phosphorylated zeta chain was digested with trypsin. Separation and identification of the zeta chain fragments using LC-MS showed, as predicted by the peptide phosphorylation studies, that tyrosine 1N is indeed the first to be phosphorylated by Lck. We conclude that differences in the amino-acid context of the six zeta chain ITAM tyrosines affect the efficiency of their phosphorylation by the kinase Lck, which probably contributes to the distinct patterns of phosphorylation observed in vivo.

Prolactin signalling in porcine theca cells: the involvement of protein kinases and phosphatases.

The hypothesis that protein kinase C (PKC) and tyrosine kinases, as well as serine-threonine and tyrosine phosphatases, are involved in prolactin (PRL) signalling in theca cells harvested from porcine follicles was tested. Theca cells were incubated with PRL for 24 h to stimulate progesterone (P4) production. In addition, treatments included inhibitors of PKC and tyrosine kinases, as well as serine-threonine phosphatase inhibitor and tyrosine phosphatase inhibitor. Prolactin significantly stimulated P4 production by theca cells and all inhibitors suppressed the PRL-stimulated P4 production. After incubation with PRL for 2, 5, 10 or 20 min, theca cells were homogenized and cytosolic and membrane fractions were obtained. This was followed by determination of PKC activity in partially purified subcellular fractions by measuring the transfer of 32P from [gamma-32P] adenosine triphosphatase (ATP) to histone III-S. In unstimulated porcine theca cells the major proportion of PKC activity was present in the cytosol. Incubation of cells with PRL resulted in a rapid, time-dependent increase in the amount of PKC activity in the membrane fraction. Protein kinase C activity in the membrane fraction was maximal after 10 min of cells' exposure to PRL. Protein kinase C activation was assessed also by measuring the specific association of 3H-phorbol dibutyrate (3H-PDBu) with theca cells after treatment with PRL. Prolactin significantly increased 3H-PDBu-specific binding in theca cells. In contrast to PKC, total inositol phosphate accumulation was not affected by PRL in the current study. In summary, PRL stimulated P4 production by porcine theca cells derived from large follicles. The results of the study were consistent with the hypothesis that PKC is one of the intracellular mediators of PRL action in porcine theca cells. Protein kinase C activation does not appear to occur through the action of phosphatidylinositol-dependent phospholipase C. Moreover, the involvement of tyrosine kinases, as well as tyrosine and serine-threonine phosphatases, in PRL signalling in the examined cells is suggested.

The effect of earthworms and arbuscular mycorrhizal fungi on growth of and 32P transfer between Allium porrum plants

To measure P transfers, two small pots, placed 1 cm apart and each containing one leek ( Allium porrum L.) plant, were embedded within a larger sand volume. Mesh windows on the facing pot sides allowed the penetration of hyphae but not roots. The effect of earthworms ( Aporrectodea caliginosa Sav.) in the hyphal compartment and of arbuscular mycorrhizal fungi (AMF) on plant growth and 32P transfer between plants was investigated. In addition, the effect of mechanical disruption of the hyphal compartment on mycorrhizal plants was studied. AMF did not affect plant growth, although infection levels were high. Earthworms generally increased shoot and root growth; this effect did not appear to be related to location of earthworm activity. 32P transfer from the dying root system of labelled ‘donor’ plants to unlabelled ‘receiver’ plants was increased where earthworm activity was evident between pots, whereas mechanical disruption eliminated the effect of AMF on 32P transfer between mycorrhizal plants. Magnitude of transfer was only related to AMF colonisation levels in the donor plant in the absence of earthworms or mechanical disruption. These results suggest that movement of 32P into the hyphal compartment via the donor AMF mycelium was more important in influencing transfer magnitude than direct transfer via AMF hyphal interconnections between the two plants. Uptake of 32P by the receiver AMF hyphae appears to have been increased by earthworm activity in the hyphal compartment; it is proposed that earthworm-mediated mobilisation of 32P, partly contained within the donor AMF mycelium, led to enhanced 32P availability in the hyphal compartment.

Effects of resource availability on mycelial interactions and 32P transfer between a saprotrophic and an ectomycorrhizal fungus in soil microcosms

Interactions between mycelia of the ectomycorrhizal fungus Suillus variegatus and the wood decomposing fungus Hypholoma fasciculare were studied in soil microcosms. The ectomycorrhizal mycelium extended from Pinus sylvestris seedlings and the saprotrophic fungus grew out into the soil from birch wood blocks of two alternative sizes. Transfer of 32P between the interacting mycelia was measured non-destructively using electronic autoradiography. The outcome of the interactions was clearly affected by the size of the wood blocks. In systems with large wood blocks (1.6 cm3) the wood decomposer fungus overgrew the mycelium of the mycorrhizal fungus whereas in systems with small wood blocks (0.44 cm3) the mycorrhizal mycelium overgrew the wood decomposer fungus. The saprotrophic fungus was able to capture significantly more 32P from labelled mycorrhizal mycelium when growing out from larger wood blocks. The mycorrhizal fungus was able to capture significantly more 32P from labelled saprotrophic mycelium when the wood blocks were smaller.

Effects of resource availability on mycelial interactions and 32P transfer between a saprotrophic and an ectomycorrhizal fungus in soil microcosms

Interactions between mycelia of the ectomycorrhizal fungus Suillus variegatus and the wood decomposing fungus Hypholoma fasciculare were studied in soil microcosms. The ectomycorrhizal mycelium extended from Pinus sylvestris seedlings and the saprotrophic fungus grew out into the soil from birch wood blocks of two alternative sizes. Transfer of 32P between the interacting mycelia was measured non-destructively using electronic autoradiography. The outcome of the interactions was clearly affected by the size of the wood blocks. In systems with large wood blocks (1.6 cm 3) the wood decomposer fungus overgrew the mycelium of the mycorrhizal fungus whereas in systems with small wood blocks (0.44 cm 3) the mycorrhizal mycelium overgrew the wood decomposer fungus. The saprotrophic fungus was able to capture significantly more 32P from labelled mycorrhizal mycelium when growing out from larger wood blocks. The mycorrhizal fungus was able to capture significantly more 32P from labelled saprotrophic mycelium when the wood blocks were smaller.

Actin is ADP-ribosylated by the Salmonella enterica virulence-associated protein SpvB.

The Salmonella enterica virulence-associated protein SpvB was recently shown to contain a carboxy-terminal mono(ADP-ribosyl)transferase domain. We demonstrate here that the catalytic domain of SpvB as well bacterial extracts containing full-length SpvB modifies a 43 kDa protein from macrophage-like J774-A.1 and epithelial MDCK cells as shown by label transfer from [32P]-nicotinamide adenine dinucleotide (NAD) to the 43 kDa protein. When analysed by two-dimensional gel electrophoresis, the same protein was modified in cells infected with S. enterica serovariant Dublin strain SH9325, whereas infection with an isogenic spvB mutant strain did not result in modification. Immunoprecipitation and immunoblotting experiments using SH9325-infected cells identified the modified protein as actin. The isolated catalytic domain of SpvB mediated transfer of 32P from [32P]-NAD to actins from various sources in vitro, whereas isolated eukaryotic control proteins or bacterial proteins were not modified. In an in vitro actin polymerization assay, the isolated catalytic SpvB domain prevented the conversion of G actin into F actin. Microscopic examination of MDCK cells infected with SH9325 revealed morphological changes and loss of filamentous actin content, whereas cells infected with the spvB mutant remained virtually unaffected. We conclude that actin is a target for an SpvB-mediated modification, most probably ADP-ribosylation, and that the modification of G actin interferes with actin polymerization.

32P mobility in detached leaf of several crop plants

Detached leaves of several crop plants were cultured in a nutrient solution containing 32P, and the 32P mobility between and within organs was investigated. The results obtained were as follows: 1) In soybean and adzuki bean, 32P was well distributed all over the leaf blade and petiole within 24 h after 32P absorption, though the transfer of 32P from the leaf vein to the mesophyll varied among the varieties. 2) In rice, 32P reached the middle or proximal part of the leaf blade within 48 h after 32P absorption. The gradient of 32P density decreased distally. 3) In maize and pumpkin, a small amount of 32P reached the leaf blade; in maize, the gradient of 32P density was similar to that in rice, though 32P was retained in the whole petiole in pumpkin. Besides transpiration, 32P transfer from xylem to mesophyll and structures of leaf tissues could be considered to regulate the 32P mobility.

Translocation of 32P between interacting mycelia of a wood‐decomposing fungus and ectomycorrhizal fungi in microcosm systems

Interactions between saprotrophic and ectomycorrhizal fungi have been largely ignored, although their mycelia often share the same microsites. The mycelial systems show general similarity to each other and, although the enzymatic potential of the saprotrophic fungi is generally considered to be higher, the importance of organic nutrient sources to ectomycorrhizal fungi is now widely accepted. In the experiments described here, nutritional interactions involving transfer of elements from one mycelium to the other have been monitored dynamically using radioactive tracers and a non‐destructive electronic autoradiography system. Microcosms were used in which mycelial systems of the ectomycorrhizal fungi Suillus variegatus and Paxillus involutus, extending from Pinus sylvestris host plants, were confronted with mycelia of the saprotroph Hypholoma fasciculare extending from wood blocks. The fungi showed a clear morphological confrontation response. The mycorrhizal mycelium often formed dense patches over the Hypholoma mycelia. Up to 25% of the 32P present in the Hypholoma mycelium was captured by the mycorrhizal fungi and translocated to the plant host within 30 d. The transfer of 32P to the saprotroph from labelled mycorrhizal mycelium was one to two orders of magnitude lower. The significance of this transfer as a ‘short cut’ in nutrient cycling is discussed.

Assay and purification of calmodulin-dependent protein kinase.

AbstractPosttranslation covalent modification of proteins by phosphorylation represents an important mechanism in the control of many cellular functions (1,2). Protein kinases catalyze the transfer of the γ-phosphoryl group of ATP to an acceptor protein substrate. The activity of the enzyme is determined by the transfer of 32P from [γ-32P] ATP to protein substrate KeywordsFilter Paper DiscGeiger Muller CounterGlass Sintered FilterPhenyl Methyl Sulfonyl FluorideDivinyl SulfoneThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Measurements of phosphorus uptake by macrophytes and epiphytes from the LaPlatte River (VT) using 32P in stream microcosms

1. Phosphorus (P) uptake by macrophytes and epiphytes from the LaPlatte River (VT) was examined in the laboratory by adding 32PO4‐P to recirculating stream microcosms.2. Water, plugs of sediment and plants were removed from the river and placed into the microcosms. 32PO4‐P was then added either to the water or the sediment, and its incorporation into plants and epiphytes was monitored over 3 days. Uptake was examined at both ambient (5 μg L–1) and increased (50 μg L–1) soluble reactive phosphorus (SRP) concentrations. A computer program was developed to fit curves to the radiotracer data and calculate rate constants for the simultaneous transfer of 32P among compartments.3. Both macrophytes and epiphytes removed P from the water, but epiphyte uptake of P was more rapid. Phosphate enrichment stimulated P uptake by both macrophytes and epiphytes. Macrophytes also obtained P from the sediment. The relative contribution of P to macrophytes from the water vs. that from the sediment appeared to vary with SRP in the overlying water. Accurate estimates of rates of P uptake from sediments by macrophytes were difficult to obtain however, due to very low and highly variable unit rate constants for P uptake and uncertainty about the magnitude of the phosphate pool available for uptake.4. SRP concentrations were greater in the overlying water than in the sediment pore water of stream microcosms in the present study. Numerous reports in the literature have suggested that this condition favours uptake by macrophyte stems and leaves rather than by roots.5. Phosphate uptake from the water by macrophytes in shallow streams may be more common than for macrophytes in lakes.

Placental transfer of phosphate in anaesthetized rats

Mechanisms of transfer of inorganic phosphate, Pi, across the placenta of rats at 21 days of gestation were studied using 32Pi. In one group of experiments the unidirectional transfer of Pi from mother to fetus was estimated from radioactivity in the fetus at various intervals after the tracer injection into the mother. At 15 min after tracer injection, the transfer rate was only slightly higher than the estimated rate of fetal accretion of Pi, and it decreased rapidly with the length of the experiment suggesting deterioration of the transfer mechanism under conditions of an acute experiment. In other experiments, transfer of 32Pi and 51Cr-EDTA (a marker of paracellular transfer) were measured across the dually-perfused placenta in the maternal-fetal direction. The transfer rate of 32Pi was an order of magnitude higher than the transfer of 51Cr-EDTA indicating that most of the Pi transfer is transcellular. The transfer of 32P decreased when the concentration of Na + in the maternal perfusate was reduced, it was related inversely to the concentration of Pi on the fetal side of the placenta, and it was related directly to the concentration of Ca 2+ on the fetal side. The maternal-fetal transfer of Pi exhibited saturation kinetics with a K m of about 0.4 mm suggesting that at a physiological concentration of Pi in maternal plasma the transfer mechanism is nearly saturated. The present observations are consistent with Pi being transferred in cotransport with Na +. The maternal-fetal transport of Pi appears to be stabilized by the high affinity of the transport system to Pi, and controlled by a negative feedback between fetal concentration of Pi and the Pi transfer rate. It may also be controlled, to some degree, by the fetal plasma Ca 2+.

Interspecific nutrient transfer in a tallgrass prairie plant community

Interplant nutrient transfer may be an important ecological process in grasslands, and may significantly influence plant neighborhood interactions. We investigated the potential for phosphorus transfer between the dominant grass Andropogon gerardii and several neighboring plant species in tallgrass prairie via a field 32PO4 labelling experiment. The mean amount of 32P received from donor shoots differed significantly among neighboring species and decreased with increasing distance from the donor. In general, forbs and cool‐season C3 grasses received more labelled 32P than warm‐season C4 grasses. Phosphorus transfer occurred over distances up to 0.5 m. The effects of species and distance on movement of phosphorus changed with increasing time after labelling. The relative mass of receiver and donor shoots did not affect amounts of 32P transfer. A benomyl fungicide treatment, applied to suppress mycorrhizal activity, likely did not affect existing vegetative hyphae and did not affect the amount of 32P transferred. These studies demonstrate that: (1) phosphorus is transferred among neighboring species in tallgrass prairie plant communities, (2) phosphorus may be transferred over significantly greater distances than reported in other grasslands, and (3) there is differential transfer among co‐occurring species. Hypothesized mechanisms accounting for these patterns in tallgrass prairie include mycorrhizal hyphal interconnections and/or extensive and differential root and rhizosphere overlap among neighboring species.