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  • Immunocytochemistry  (41)
  • Rhizobium  (29)
  • Springer  (70)
  • Blackwell Publishing Ltd
  • 1990-1994
  • 1980-1984  (70)
  • 1925-1929
  • 1984  (70)
Collection
Publisher
Years
  • 1990-1994
  • 1980-1984  (70)
  • 1925-1929
Year
  • 1
    ISSN: 1432-041X
    Keywords: Major haemolymph proteins ; Development ; Cuticle ; Immunocytochemistry ; Ceratitis capitata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The developmental profile of the major haemolymph proteins (ceratitins) inCeratitis capitata was studied. Ceratitin concentration in the haemolymph decreases dramatically during the last days of pupal life, while the amounts of ceratitins in whole organism extracts remain unchanged. By electrophoretic, immunological and immunofluorescence techniques it was revealed that ceratitins are reabsorbed by the fat body and a fraction of them is deposited in the cuticle. The possible role of ceratitins is discussed.
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  • 2
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    Planta 161 (1984), S. 32-36 
    ISSN: 1432-2048
    Keywords: Ononitol ; O-Methyl-scyllo-inositol ; Pisum (root nodule) ; Rhizobium ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ononitol (4-O-methyl-myo-inositol) and O-methyl-scyllo-inositol were identified in pea (Pisum sativum L.) root nodules formed by twoRhizobium leguminosarum strains. Ononitol was the major soluble carbohydrate in nodules formed by strain 1045 while O-methyl-scyllo-inositol and two unidentified components were dominant in the carbohydrate pattern of the nodules formed by strain 1 a. The cyclitols were also present in the denodulated roots, but to a much smaller extent; in the above-ground plant parts only traces were found. The identification of ononitol and O-methyl-scyllo-inositol was established by gas chromatography and gas chromatography-mass spectrometry utilizing trimethylsilyl- and acetyl-derivatives.
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  • 3
    ISSN: 1432-2048
    Keywords: Bacteroid ; Glycine (bacteroids) ; Denitrification ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrate, nitrite and nitrous oxide were denitrified to N2 gas by washed cells ofRhizobium japonicum CC706 as well as by bacteroids prepared from root nodules ofGlycine max (L.) Merr. (CV. Clark 63). Radiolabelled N2 was produced from either K15NO3 or Na15NO2 by washed cells ofRh. japonicum CC705 grown with either nitrate only (5 mM) or nitrate (5 mM) plus glutamate (10 mM). Nitrogen gas was also produced from N2O. Similar results were obtained with bacteroids ofG. max. The stoichiometry for the utilization of15NO 3 - or15NO 2 - and the produciton of15N2 was 2:1 and for N2O utilization and N2 production it was 1:1. Some of the15N2 gas produced by denitrification of15NO 3 - in bacteroids was recycled via nitrogenase into cell nitrogen.
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  • 4
    ISSN: 1432-2048
    Keywords: Immunocytochemistry ; Lectin (localization) ; Phaseolus (lectin) ; Phytohemagglutinin ; Seed (lectin)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have examined the properties and subcellular localization of phytohemagglutinin (PHA), the major lectin of the common bean (Phaseolus vulgaris.), in the axis cells of nearly mature and imbibed mature seeds. On a protein basis the axis contained about 15% as much PHA as the cotyledons. Localization of PHA was done with an indirect immunolabeling method (rabbit antibodies against PHA, followed by colloidal gold particles coated with goat antibodies against rabbit immunoglobulins) on ultra-thin cryosections which were embedded in plastic on the grids after the immunolabeling procedure. The embedding greatly improved the visualization of the subcellular structures. The small (4 nm) collodial gold particles, localized with the electron microscope, were found exclusively over small vacuoles or protein bodies in all the cell types examined (cortical parenchyma cells, vascular-bundle cells, epidermal cells). The matrix of these vacuoles-protein bodies appears considerably less dense than that of the protein bodies in the cotyledons, but the results confirm that in all parts of the embryo PHA is localized in similar structures.
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  • 5
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    Archives of microbiology 137 (1984), S. 124-127 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Osmotic stress ; Salts ; Glutamic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A defined medium of low osmolarity was developed permitting growth of Rhizobium meliloti with generation times of approximately 2.8 h doubling-1. The effects of sodium, potassium, magnesium, ammonium, chloride, sulfate, phosphate, bicarbonate and acetate ions on the growth rate of R. meliloti were determined. Sodium, potassium and ammonium ions had little effect on growth at concentrations of 100 mEq or less; magnesium ion inhibited growth severely at concentrations of 50 mEq (25 mM). Of the anions, chloride and sulfate appeared to have little effect while phosphate, bicarbonate, and acetate inhibited growth at concentrations of as little as 25 mEq. The addition of proline, glutamate, or betaine to cells growing in inhibitory concentrations of NaCl did not relieve the inhibition. When grown in the presence of inhibitory levels of NaCl, the intracellular concentration of glutamate but not of proline or gamma amino butyric acid increased 5-fold.
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  • 6
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    Archives of microbiology 140 (1984), S. 281-286 
    ISSN: 1432-072X
    Keywords: Alkaline phosphatase ; Continuous culture ; Glycerol 1-phosphate uptake ; Phosphate exchange ; Phosphate uptake ; Rhizobium ; Snake bean bacteroids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of P nutrition on phosphate uptake and alkaline phosphatase activity was studied in chemostat culture for four rhizobial and three bradyrhizobial species. Phosphate-limited cells took up phosphate 10- to 180-fold faster than phosphate-rich cells. The four fast-growing rhizobial strains contained high levels of alkaline phosphatase activity under P-limited conditions compared to the repressed levels found in P-rich cells; alkaline phosphatase activity could not be detected in three slow-growing rhizobial strains, regardless of their P-status. Glycerol 1-phosphate-uptake in the cowpea Rhizobium NGR234 was derepressed over 50-fold under P-limited conditions, and appeared to be co-regulated with phosphate uptake. The phosphate-uptake system appeared similar in all strains with apparent K m values ranging from 1.6 μM to 6.0 μM phosphate and maximum activities from 17.2 to 126 nmol · min-1 · (mg dry weight of cells)-1. Carbonyl cyanide m-chlorophenyl hydrazone strongly inhibited phosphate uptake in all strains and a number of other metabolic inhibitors also decreased phosphate uptake in the cowpea Rhizobium NGR234. The phosphate uptake system in all strains failed to catalyse exchange of 32P label in preloaded cells or efflux of phosphate. The results suggest a single, repressible, unidirectional and energy-dependent system for the transport of phosphate into rhizobia.
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  • 7
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    Archives of microbiology 138 (1984), S. 187-190 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Aromatic metabolism in protocatechuate 3,4-dioxygenase ; R. trifolii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protocatechuate 3,4-dioxygenase (EC 1.13.11.3) has been purified 42-fold from 4-hydroxybenzoate-grown cells of Rhizobium trifolii TA1, where it constitutes about 2% of the cell protein. The dioxygenase has a molecular weight of 220,000, with two dissimilar sub-units of molecular weights 29,000 and 26,500, corresponding to an α4β4 composition. The enzyme is specific for protocatechuate, with a Km of 1.75×10-5 M and maximum activity at pH 9.2. Metal removal and replacement studies indicate that the enzyme contains complexed Fe3+ which is required for activity. Direct atomic absorption analysis gave 1.3–1.5 g atoms Fe3+ per mole of isolated enzyme, but correction for metal-deficient proteins suggests that the value is close to 2.
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  • 8
    ISSN: 1432-072X
    Keywords: Rhizobium ; Nitrogenase ; Hydrogenase ; Growth yields ; Chemostat ; Cytochromes ; Proton translocation ; ATP/N2 ratio
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rhizobium ORS 571, isolated from stem nodules of the tropical legumeSesbania rostrata is able to grow in the chemostat with molecular nitrogen as sole nitrogen source at a specific growth rate of 0.1 h-1. Samples from nitrogenfixing cultures showed high acetylene reduction activities: 1,500 nmol ethylene formed per milligram dry weight per hour. Under nitrogen-fixing conditions an uptake hydrogenase is induced. Ammonia-assimilating cultures, without additional hydrogen, did not induce hydrogenase. The addition of hydrogen to succinate-limited nitrogen-fixing cultures resulted in an increase in the molar growth yield on succinate (Y succinate) from 27 to 35 and a slight decrease in the molar growth yield on oxygen ( $$Y_{O_2 }$$ ), showing that hydrogen oxidation is less energy-yielding than the oxidation of endogenous substrates. Respiration-driven proton translocation measured with starved cells indicated the functioning of site 1 and 2 of oxidative phosphorylation. Cytochrome spectra showed that cytochromea 600, present at high dissolved oxygen tension (d.o.t.) almost completely disappeared at low d.o.t. In flash-photolysis spectra only thea-type cytochrome could be detected as an oxidase in cells both grown at high and low d.o.t. Growth yields in ammonia-assimilating cultures were higher than those measured in nitrogen-fixing cultures. Assuming two sites of oxidative phosphorylation, a molar growth yield on ATP (Y ATP) of about 3 and 6 was calculated for respecticely nitrogen-fixing and ammonia-assimilating cultures. TheY ATP under nitrogen-fixing conditions is dependent on the amount of H2 formed per mol N2 fixed (H2/N2 ratio). A method has been described to calculate the total amount of ATP use by nitrogenase during the fixation of 1 mol N2 (ATP/N2 ratio) and H2/N2 ratios in aerobic nitrogen fixing organisms. This calculation yielded that nitrogen fixation inRhizobium ORS 571 is a high ATP-consuming process. The calculated ATP/N2 and H2/N2 ratios were respectively 42 and 7.5.
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  • 9
    ISSN: 1432-072X
    Keywords: Alkaline phosphatase ; Continuous culture ; Growth yields ; Phosphate ; Phosphorus nutrition ; Polyphosphate ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract With continuous cultures in a fully defined minimal salts medium steady states were achieved at both limiting and non-limiting concentrations of phosphate in the inflowing medium for Rhizobium trifolii WU95, cowpea Rhizobium NGR234, and Bradyrhizobium CB756. Millimolar growth yields obtained from P-limited cultures varied over 2-fold from 3.2 g dry weight·(mmol P)-1 for WU95 to 5.3 g dry weight·(mmol P)-1 for CB756 and 7.2 g dry weight·(mmol P)-1 for NGR234. For both WU95 and NGR234 growth under P-excess conditions resulted in elevated levels of total biomass P and the storage compound polyphosphate, compared with P-limited cultures. However, P-limited cultures of these two strains still contained significant quantities of polyphosphate. The P-status for CB756 cultures did not affect either total biomass P or polyphosphate levels. Alkaline phosphatase was maximally derepressed in P-limited cultures of WU95 and NGR234. However, in CB756 alkaline phosphatase was not detected at significant levels regardless of its P supply. These data suggest that growth of rhizobia is controlled predominantly by the attainment of a critical internal P level.
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  • 10
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    Archives of microbiology 140 (1984), S. 260-264 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Legume ; Preinfection events ; Symbiont recognition ; Capsular polysaccharide composition ; Root exudate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The capsular polysaccharides (CPS) of the log phase cells of Rhizobium sp. strain 1001 were altered within 6 h of incubation in the root exudate of cowpea (Vigna sinensis) plants grown in the absence of combined nitrogen. Electrophoretic or DEAE-Sephadex separation of the native CPS yielded two fractions while the CPS of the cultures incubated in root exudate (host induced CPS) yielded three fractions. The relative proportions of sugars in the total CPS were different. The host induced CPS contained two new sugars, arabinose and xylose, in addition to the native components of mannose, glucose and galactose. Addition of the host induced CPS to cowpea roots enhanced the nodulation efficiency of Rhizobium sp. strain 1001 under low inoculum density. None of the above host mediated changes occurred on incubation of the rhizobia with root exudate of cowpea plants grown in the presence of NH 4 + , indicating the regulatory effect of combined nitrogen on the symbiotic properties of legume root secretions.
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  • 11
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    Archives of microbiology 140 (1984), S. 287-290 
    ISSN: 1432-072X
    Keywords: Alkaline phosphatase ; Continuous culture ; Periplasmic proteins ; Phosphate uptake ; Phosphorus nutrition ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Lysozyme/EDTA treatment of four fast-growing rhizobia released repeatable protein profiles after polyacrylamide slab gel electrophoresis. Similar treatment of slow-growing rhizobia failed to release such periplasmic proteins. For the four-fast-growing rhizobia, both P-repressible and P-inducible protein bands occurred. The only P-repressible protein identified was alkaline phosphatase, which showed strain differences in both electrophoretic mobility and activation by Mg2+. The derepression of the P-repressible periplasmic proteins in cowpea Rhizobium NGR234 correlated with derepression of both phosphate and glycerol 1-phosphate uptake.
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  • 12
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    Plant molecular biology 3 (1984), S. 21-28 
    ISSN: 1573-5028
    Keywords: soybean ; nitrogen fixation ; nodule-specific genes ; Rhizobium ; Northern blots ; RNA dot blots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cloned cDNAs corresponding to mRNAs which accumulate in nitrogen-fixing root nodules of soybean (nodulin mRNAs) were used as probes to investigate the sizes, sequence relationships, tissue specificities and developmental accumulations of individual nodulin mRNA sequences. Northern blot analysis indicated that the NodB, NodC and NodD mRNA sequences are 1 150, 770, and 3 150 nucleotides long, respectively, which is consistent with the previously determined sizes of the hybrid-selected translation products (27 000, 24 000 and 100 000 MW, respectively). The NodA clones pNodA15 and pNodA25 hybridized to two mRNAs of lengths 1 600 and 1 100 nucleotides, indicating that they contain significant sequence homologies. However, increasing the hybridization stringency showed that the pNodA15 clone encodes the 1 600 nucleotide mRNA corresponding to the major NodA hybrid-selected translation product (44 000 MW) while pNodA25 encodes an mRNA of 1 100 nucleotides. The latter probably corresponds to one of two smaller (23 500 and 24 500 MW) in vitro translation products. RNA dot-blot hybridizations indicated that nodulin and leghemoglobin mRNAs began to appear and accumulate in Rhizobium infected root tissue very early (day 3 to 5) and reached fully induced levels by day 11. This accumulation was specific for nodule tissue (except for the NodD sequence) and preceded the accumulation of nitrogen fixation activity. Nodules produced by different effective Rhizobium strains accumulated similar levels of leghemoglobin and nodulin mRNAs while ineffective strains had a pleiotropic affect. While one ineffective strain (61A24) gave reduced levels of all these mRNAs, the other (SM5) gave levels which were nearly normal by the time nitrogen fixation activity should have reached its maximal level (day 17). Thus, leghemoglobin and nodulin genes are switched on soon after infection, prior to nodule morphogenesis, and the switch occurs prior to and is independent of nitrogen fixation activity.
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  • 13
    ISSN: 1573-5028
    Keywords: electron microscopy ; mutants ; nitrogen fixation ; Rhizobium ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Insertion and deletion mutants were used to characterize a genomic region of Rhizobium japonicum where the nitrogenase structural genes are located on two separate operons nifDK and nifH. In addition to previously described nifD:: Tn5 and nifK:: Tn5 mutations we have now generated, by localized mutagenesis, further Tn5 insertion mutations in the vicinity of nifDK as well as within and adjacent to nifH. The nifD:: Tn5, nifK:: Tn5, and nifH:: Tn5 mutant strains were of the Nod+ Fix- phenotype whereas all other mutants were symbiotically fully effective (Nod+ Fix+). The nifH:: Tn5 mutation was helpful in the identification of the nifH gene product (the dinitrogenase reductase) by two-dimensional gel electrophoresis: due to its polar effect this insertion specifically abolished the synthesis of that protein under microaerobic culture conditions. The ultrastructure of soybean root nodules infected with either the nif + wild-type or with the nif - (but otherwise isogenic) mutant strains was analyzed by electron microscopy. All contained fully developed bacteroids, but the nitrogen non-fixing mutants showed massive accumulation of PHB. Of Tn5-containing strains, kanamycin sensitive derivatives were obtained which contained deletions. Several classes of deletion mutants were found which, as judged by their physical DNA structure and their phenotypes, allowed the following most important conclusions: (i) deletions lacking both the nifDK and nifH regions indicate linkage between the two operons whereby at least 15 kb of DNA separate them; (ii) one deletion ending upstream from nifH, and lacking only nifDK, indicates that the nifDK operon is located on the 5′-flanking side of the nifH operon; (iii) all deletion mutants are Nod+ indicating that there are no essential nodulation gnes located between and adjacent to nifDK and nifH.
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  • 14
    ISSN: 1432-2048
    Keywords: Lectin ; Nitrate and lectin formation ; Rhizobium ; Trifoliin A ; Trifolium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In-vivo synthesis of the white-clover lectin, trifoliin A, was examined by the incorporation of labeled amino acids into protein during heterotrophic growth of intact Trifolium repens L. seedlings. Lectin synthesis was quantified by measuring the level of labeled protein immunoprecipitated from root exudate, from the hapten (2-deoxyglucose) eluate of the roots, and from root and shoot homogenates. The presence of labeled trifoliin A was confirmed by non-denaturing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by fluorography and comparison with trifoliin A standards. In-vivo-labeled trifoliin A was detected in seedling root homogenate 2 h after the addition of labeled amino acids and on the root surface by 8 h. Incorporation of labeled amino acids into protein and trifoliin A was greatest with 2-d-old seedlings and was greater when the plants were grown continuously in the dark than when they were exposed to 14 h light daily. Significantly more labeled lectin accumulated on the root surface of seedlings grown with 1.5 mM KNO3 than of seedlings grown either without N or with 15.0 mM KNO3. The labeled lectin from the root surface in all nitrate treatments and from the rootexudate samples of seedlings grown N-free and with 1.5 mM KNO3 was fully able to bind to Rhizobium trifolii. In contrast, only 2% of the immunoprecipitable protein found in the root exudate of seedlings grown with 15.0 mM KNO3 was able to bind to the bacteria. Thus, excess nitrate does not repress the synthesis of trifoliin A in the root, but does affect the distribution and activity of this newly synthesized lectin in a way which reduces its ability to interact with R. trifolii. By using Western blot analysis, much more total trifoliin A is detected in the homogenates of shoots than roots. However, greater than 80% of the total labeled protein and 85–90% of the total labeled lectin were found in the root homogenates of 2-d-old dark-grown seedlings incubated for 5 h with labeled amino acids. In addition, Western blot analysis indicated that the shoot homogenate contained smaller-molecular-weight peptides which reacted with the specific anti-trifoliin A antibody. These studies indicate that stored trifoliin A in the seed is degraded in the shoots during seedling development, while newly synthesized trifoliin A in the roots is excreted to the root surface and external environment.
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  • 15
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    Cell & tissue research 237 (1984), S. 195-202 
    ISSN: 1432-0878
    Keywords: Pituitary ; Prolactin cells ; Estrogen ; Heterogeneity ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study was conducted to determine the functional and/or developmental relationships among three heterogeneous types of prolactin cells (I, II and III) in rats. Rats were injected subcutaneously daily with estradiol or testosterone propionate on days 10–20 after birth. Estradiol increased the proportion of cell types II and III, increased serum PRL levels 12-fold in males and 15-fold in females, and increased pituitary levels of prolactin 12-fold in males and 5-fold in females. Testosterone mainly increased the proportion of the Type-II cells, decreased serum levels of prolactin in males only, and did not change pituitary levels of prolactin. In a second experiment, treatment of rats with nafoxidine for five days after E2 treatment (days 10–20 after birth) increased the proportion of Type-I cells and decreased the proportion of Type-III cells and decreased serum and pituitary levels of prolactin by 50% in females and by 15 and 45% in males. In a third experiment utilizing adult male rats, estradiol and testosterone were found to modulate the relative ratios of the different types of PRL cells as they did in immature animals. The data taken as a whole suggest the possibility of an estrogen-stimulated conversion of one cell type to another, which may be a reflection of prolactin secretory activity.
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  • 16
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    Cell & tissue research 237 (1984), S. 245-252 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Interstitial cells ; Astrocytes ; Immunocytochemistry ; Rat ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antigenic markers characteristic of astrocytes and their differentiative states (i.e., glial fibrillary acidic protein (GFAP), vimentin, and M1 and C1 antigens) were investigated in the pineal gland of mouse and rat using double immunolabeling techniques. In both species the socalled interstitial cells as characterized by TEM were shown to be astrocytes, since they expressed vimentin, but neither fibronectin (a marker for fibroblasts and endothelial cells) nor the neuron-specific L1 antigen or tetanus toxin receptors. Subpopulations of vimentin-positive pineal astrocytes were also GFAP- and C1- antigen-positive. M1- antigenpositive cells were not detected. It is concluded that a considerable proportion of interstitial cells in the pineal gland of rat and mouse are immature astrocytes which, in contrast to other parts of the central nervous system, persist into adulthood.
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  • 17
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Reissner's fiber ; Ependyma ; Secretory process ; Comparative analysis ; Immunocytochemistry ; Vertebrates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organs (SCO) of 76 specimens belonging to 25 vertebrate species (amphibians, reptiles, birds, mammals) were studied by use of the immunoperoxidase procedure. The primary antiserum was obtained by immunizing rabbits with bovine Reissner's fiber (RF) extracted in a medium containing EDTA, DTT and urea. Antiserum against an aqueous extract of RF was also produced. The presence of immunoreactive material in cell processes and endings was regarded as an indication of a possible route of passage. Special attention was paid to the relative development of the ventricular, leptomeningeal and vascular pathways established by immunoreactive structures. The SCO of submammalian species is characterized by (i) a conspicuous leptomeningeal connection established by ependymal cells, (ii) scarce or missing hypendymal cells, and (iii) a population of ependymal cells establishing close spatial contacts with blood vessels. The SCO of most mammalian species displays the following features: (i) ependymal cells lacking immunoreactive long basal processes, (ii) hypendymal secretory cells occurring either in a scattered arrangement or forming clusters, (iii) an occasional leptomeningeal connection provided by hypendymal cells, and (iv) in certain species numerous contacts of secretory cells with blood vessels. In the hedgehog immunoreactive material was missing in the ependymal formation of the SCO, but present in hypendymal cells and in the choroid plexuses. The SCO of several species of New-and Old-World monkeys displayed immunoreactive material, whereas that of anthropoid apes (chimpanzee, orangutan) and man was completely negative with the antisera used.
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  • 18
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Ependyma ; Comparative aspects ; Immunocytochemistry ; Secretory process ; Blood vessels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In 76 specimens (amphibians, reptilians, mammals) belonging to 25 different vertebrate species, the region of the subcommissural organ (SCO) was investigated with the use of a primary antiserum raised against an extract of bovine Reissner's fiber+the immunoperoxidase procedure according to Sternberger et al. (1970). In the SCO of a toad (Bufo arenarum) and several species of reptiles (lacertilians, ophidians, crocodilians), the ependymal cells were the only type of secretory cell displaying vascular contacts, whereas in mammals ependymal and hypendymal cells established intimate spatial contacts with blood vessels. In Bufo arenarum, but especially in the reptilian species examined, the ependymo-vascular relationship was exerted by a population of ependymal cells having a rather constant location within the SCO and projecting to capillaries that showed a remarkably constant pattern of anatomical distribution. In the SCO of mammals the modality and degree of the structural relationships between secretory cells and blood vessels varied greatly from species to species. In the SCO of the armadillo and dog the secretory tissue was organized as a thick, highly vascularized layer with most of the cells oriented toward the capillaries. A rather opposite situation was found in the SCO of New-and Old-World monkeys, where vascular contacts were restricted to a few ependymal cells.
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  • 19
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    Cell & tissue research 237 (1984), S. 521-524 
    ISSN: 1432-0878
    Keywords: α-Melanocyte-stimulating hormone ; α-MSH-like peptide ; Immunocytochemistry ; Hypothalamus ; Lizard (Lacerta muralis)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of α-melanocyte-stimulating hormone (α-MSH) was studied in the brain of the lizard Lacerta muralis by means of immunocytochemical staining methods. α-MSH-like containing cells were found in the ventro-lateral preoptic area and the paraventricular and supraoptic nuclei. Some scattered cells staining for α-MSH were also detected in the mesencephalo-diencephalic boundary region, while numerous α-MSH-like nerve fibres were localized in the medial eminence. No reaction was observed after the use of antiserum preabsorbed with synthetic antigen. These findings suggest that an α-MSH-like peptidergic system could possibly be involved in the hypothalamo-hypophysial regulation and/or play a role as neurotransmitter in this animal.
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  • 20
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    Cell & tissue research 238 (1984), S. 203-205 
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Hypothalamus, human ; Immunocytochemistry ; Pituitary stalk
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In order to study the distribution of neuropeptide Y-like immunoreactivity in the human hypothalamus, an immunocytochemical localization of this peptide was performed. Using antibodies developed against synthetic porcine neuropeptide Y (NPY), we have been able to localize immunoreactivity in neuronal cell bodies located exclusively in the infundibular nucleus. Immunostained fibers were found in several regions in the hypothalamus with a high concentration in the periventricular areas. Fibers were also found in the neurovascular zone of the median eminence, the pituitary stalk and the posterior pituitary. These results suggest that immunoreactive material related to porcine NPY is present in the human hypothalamus, with a distribution similar to that observed in the rat.
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  • 21
    ISSN: 1432-0878
    Keywords: Peptidergic neurotransmission ; Lymnaea stagnalis ; Immunocytochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Three neuronal systems of the pond snail Lymnaea stagnalis were immunocytochemically investigated at the ultrastructural level with the unlabeled peroxidase-antiperoxidase technique. Preliminary electrophysiological and cell-filling investigations have shown that a cluster of neurons which reacts positively with an antiserum against the molluscan cardio-active peptide FMRFamide, sends axons to the penis retractor muscle. In this muscle anti-FMRF-amide (aFM) positive axons form neuro-muscular synapses with (smooth) muscle fibers. The morphological observations suggest the aFM immunoreactive system to be involved in peptidergic neurotransmission. In the right parietal ganglion a large neuron (LYAC) is penetrated by aFM positive axons which form synapse-like structures (SLS) with the LYAC. The assumption that the SLS represent the morphological basis for peptidergic transmission is sustained by the observation that iontophoretical application of synthetic FMRFamide depolarizes the LYAC. The axons of a group of pedal anti-vasopressin (aVP) positive cells run in close vicinity to the cerebral ovulation (neuro-)-hormone producing cell system (CDC system) Synapses or SLS between the two systems were not observed. The fact that (bath) application of arg-vasopressin induces bursting in the CDC, may indicate that the vasopressin-like substance of the aVP cells is released non-synaptically.
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  • 22
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    Cell & tissue research 238 (1984), S. 497-502 
    ISSN: 1432-0878
    Keywords: D2 glycoprotein ; Adrenal gland ; Immunocytochemistry ; Ultrastructure ; Cell adhesion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of the glycoprotein D2 in rat adrenal gland was investigated using immunohistochemical methods, and D2 localization in cultures of adult bovine chromaffin cells was studied by immunofluorescence. D2 was found to be situated on nerve fibers passing through the adrenal cortex and in the medulla zone, and also on the surface of all chromaffin cells. In addition, it was strongly expressed on the surface of glial (Schwann) cells. Cortical cells were unreactive to the antiserum. In cultures, all adrenalin and noradrenalin [dopamine-β-hydroxylase (DBH)-positive] cells were surface labelled for D2. A less frequent second cell type was recognized in vitro which was DBH negative but D2 positive. Such cells were presumed to be Schwann cells. These data are discussed in terms of the developmental origin of the cells and with regard to the putative functional rôle of D2 in cell adhesion phenomena.
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  • 23
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    Cell & tissue research 235 (1984), S. 211-214 
    ISSN: 1432-0878
    Keywords: Adenohypophysis ; Pars distalis ; Immunocytochemistry ; Amphibia ; Gymnophiona
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The indirect immunofluorescence method was used to identify and locate LTH-, STH-, LH-, TSH-, ACTH- and MSH-immunoreactive cells in the pituitary of Typhlonectes compressicaudus (Gymnophiona). The present study defines the histological and histochemical staining properties of each cell type identified.
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  • 24
    ISSN: 1432-0878
    Keywords: Caldesmon ; Actin ; Immunocytochemistry ; Small intestine ; Smooth muscle ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin-interacting protein) (Sobue et al. 1982) and of actin was studied in the rat's small intestine by means of light-microscopic immunocytochemistry. Positive immunostaining for caldesmon was seen in smooth muscle cells of the intestinal wall, and of blood vessels, and in the apical portion of the absorptive epithelial cells. The immunoreactivity in goblet cells was difficult to recognize. The positive reaction to immunostaining for actin showed almost the same pattern as that for caldesmon. These results suggest that this calmodulin-binding protein may play an important role in the control of actin-myosin interaction in smooth muscle cells and in non-muscle cells.
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  • 25
    ISSN: 1432-0878
    Keywords: FMRFamide ; Bovine pancreatic polypeptide ; Immunocytochemistry ; Peptidergic neurons ; Leptinotarsa decemlineata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Particular neurons in the nervous system of the Colorado potato beetle, Leptinotarsa decemlineata, are recognized by antisera against bovine pancreatic polypeptide and FMRFamide. Both antisera react with the same neurons. Solid phase absorptions showed that antiserum against bovine pancreatic polypeptide cross-reacts with FMRFamide, whereas antiserum against FMRFamide cross-reacts with bovine pancreatic polypeptide. Some of the immunoreactive neurons have axons branching extensively within the neuropile, which suggests that the peptide is used as transmitter. In the corpus cardiacum, a neurohaemal organ in insects, numerous immunoreactive axon terminals are present. Here, the peptide material is presumably released as a hormone.
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  • 26
    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor ; Immunocytochemistry ; Hypothalamus ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The corticotropin-releasing factor (CRF)-containing neurons were investigated in the brain of the domestic fowl by means of the peroxidase-antiperoxidase technique at the light-microscopic level. The detection of CRF-immunoreactivity was facilitated by silver intensification. CRF-containing perikarya were found in the paraventricular, preoptic and mammillary nuclei of the hypothalamus and in some extrahypothalamic areas (nuclei dorsomedialis and dorsolateralis thalami, nucleus accumbens septi, lobus parolfactorius, periaqueductal gray of the mesencephalon, nucleus oculomotorius ventralis). Immunoreactive nerve fibers and terminals were demonstrated in the external zone of the median eminence and the organum vasculosum of the lamina terminalis. These results indicate that an immunologically demonstrable CRF-neurosecretory system also exists in the avian central nervous system.
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  • 27
    ISSN: 1432-0878
    Keywords: Thyroid ; Immunocytochemistry ; Caldesmon ; Actin ; Endocytosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin interacting protein; Sobue et al. 1982) and actin was studied in the rat thyroid gland by means of light-microscopic immunocytochemistry, and the fine-structural distribution of actin filaments was examined by use of heavy meromyosin (HMM). Caldesmon and actin were demonstrated in the apical cytoplasm of almost all the follicle epithelial cells in normal as well as TSH-treated animals. Immunoreactivities for both caldesmon and actin showed almost the same pattern in localization. The smooth muscle cells of the blood vessels were also positive for caldesmon and actin. By electron microscopy, numerous actin filaments decorated by HMM and running perpendicularly or randomly to the apical surface were recognized in the apical cytoplasm of the follicle epithelial cell. These results suggest that caldesmon and actin, in conjugation with calmodulin, play a role in the regulation of cellular activity such as exocytosis and endocytosis in the apical portion of the follicle epithelial cell.
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  • 28
    ISSN: 1432-0878
    Keywords: Urotensin ; Caudal neurosecretory system ; Immunocytochemistry
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    Notes: Summary The localization of urotensin I has been investigated in the caudal neurosecretory system of the white sucker (Catostomus commersoni). The peptide is present in all the cells of the system both large and small, in the large axons passing to the urophysis, and in fine beaded fibres not only within the urophysis but also in a fine plexus lateral to the large cells in the spinal cord proper. The possibility that the caudal neurosecretory system is not a functionally uniform system but rather a collection of dissimilar cells of different synaptic inputs with a common entity, urotensin I, is discussed. Moreover, the feasibility of a urotensin I feedback loop is described.
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  • 29
    ISSN: 1432-0878
    Keywords: Pituitary rat ; LH cells ; FSH cells ; ACTH cells ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of immunocytochemistry, the development of ACTH-, LH- and FSH cells was examined in the anterior pituitary of 5-day-old neonatal, 15-day-old immature and adult rats. ACTH-positive cells are angular and the periphery of these cells is strongly reactive with anti-ACTH serum. In contrast, LH- and FSH-immunopositive cells are ovoid elements, ranging in cell size and intensity of staining. Angular cells, in which only the cell periphery reacted with anti-LHβ serum, were observed in neonatal and immature rats; however, these cells were not stained with either anti-FSHβ serum or anti-ACTH serum. Observation of serial semithin sections revealed that ACTH-immunopositive cells do not react with either anti-LHβ or anti-FSHβ serum. Finally, it was observed that ACTH cells and LH cells are both functionally differentiated already in 5-day-old neonatal rats.
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  • 30
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    Cell & tissue research 235 (1984), S. 77-83 
    ISSN: 1432-0878
    Keywords: Pituitary (rat) ; LH cells ; FSH cells ; Rapid freeze-substitution ; Immunocytochemistry ; Ferritin antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rapid freeze-substitution fixation was employed in immunocytochemical studies on the localization of LH and FSH in the typical gonadotrophs of the anterior pituitary in the untreated male rat; a modification of a recently described ferritin antibody method (Inoue et al. 1982) was used in these studies. It was shown that rapid freeze-substitution fixation provides good preservation not only of the ultrastructure but also of the antigenicity. Both LH and FSH were clearly demonstrated in the same gonadotrophic cells, but the subcellular localization of these gonadotrophins differed: (i) LH was mainly located in small secretory granules, 250–300 nm in diameter; (ii) FSH was mainly present in large secretory granules, up to 500 nm in diameter. In the pituitary gland of the adult male rat, all gonadotrophs that react to antibodies against gonadotrophins are characterized by small and large secretory granules. Other types of cells of the anterior pituitary containing either small secretory granules or resembling corticotrophs with secretory granules assembled at cell periphery did not react to either anti-LH beta or anti-FSH beta serum. For light microscopy, the peroxidase antibody method was used. All of the gonadotrophin-positive cells contain both LH and FSH. None of the pituitary cells reacted to antibody against only one gonadotrophin. However, some cells are “LH-rich” while other cells are “FSH-rich”.
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  • 31
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    Cell & tissue research 235 (1984), S. 159-169 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Gonadotropic cells ; Immunocytochemistry ; Testosterone binding ; Cryo-ultramicrotomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Several attempts have been made to localize steroids by means of immunocytological techniques. However, these methods were found inadequate for detecting steroids bound to their receptors. To localize endogenous testosterone (T) in its target cells at the ultrastructural level, an immunocytological technique was performed on ultrathin sections obtained by cryo-ultramicrotomy. T was detected in the pituitary glands obtained from intact male or female rats and castrated rats, but not in castrated + adrenalectomized rats. Animals were also injected either with testosterone, with other steroids (estradiol, progesterone, corticosterone) or with an androgen antagonist (cyproterone acetate). In addition, some ultrathin sections were preincubated either with phosphate buffers of various pH, corticosterone, cyproterone acetate solution, or with T solution. The content of T in the pituitary before and after fixation was measured by radioimmunoassay; it decreased after fixation. T immunoreactivity was localized in the gonadotropic cells only, both in the male and female rats. At the subcellular level, the immunoreactivity was detected in the cytoplasmic matrix and in the nucleus. Immunoreactive T disappeared 1) in rats after castration+adrenalectomy; by means of radioimmunoassay no T was measured in these pituitary glands; 2) in rats injected with 25 (μg/rat of cyproterone acetate; 3) after preincubation of pituitary sections on a drop of cyproterone acetate (1 × 10-6 M). The immunocytological reaction was not modified when the rats were injected with estradiol, progesterone or corticosterone (1 mg/rat), or after preincubation of the sections with corticosterone (1 × 10-3 M), or a buffer solution at pH 7.6. Lower or higher pH values led to a strong decrease in the immunoreactivity. After injection of T (15 μg/rat) the immunocytological reaction was more abundant in the nucleus and less in the cytoplasm. The immunoreactivity was again observed when the sections were preincubated with cyproterone acetate solution and then with T solution. These data suggest that T can be detected by means of immunocytochemistry. It is probably bound to a specific binding site.
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  • 32
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Supraoptic nucleus (SON) ; Oxytocin neurones ; Neuronal appositions
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    Topics: Biology , Medicine
    Notes: Summary Pre-embedding immunocytochemistry was performed on vibratome sections of the hypothalamus of lactating rats using antiserum directed against oxytocin. Electron microscopy revealed that numerous immunopositive somata and dendrites in the supraoptic nucleus were in direct apposition, without glial interposition; a number of them were also bridged by “double” synapses. The observations support the contention that the ultrastructural reorganisation of the nucleus apparent at lactation affects the magnocellular neurones secreting oxytocin.
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  • 33
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    Cell & tissue research 235 (1984), S. 427-432 
    ISSN: 1432-0878
    Keywords: DPP IV ; Salivary glands ; Ultrastructural localization ; Immunocytochemistry ; PAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of dipeptidyl peptidase IV (DPP IV) (EC 3.4.14.5) in rat submandibular and parotid glands was studied immunocytochemically by the peroxidase-antiperoxidase (PAP) method, using a monospecific antiserum against rat kidney DPP IV. There were no differences in the immunocytochemical localization of DPP IV between submandibular and parotid glands. In these glands, DPP IV was primarily found to be associated with the luminal and intercellular canalicular plasma membranes of acinar cells and with the luminal plasma membranes of intercalated and striated duct cells. Occasionally, immunoreaction of DPP IV was detected in cytoplasmic vesicles (vacuoles), lysosomes, and multivesicular bodies in some acinar cells as well as in ductal epithelial cells. Furthermore, the reaction product was also found within the lumina of peri-acinar and peri-ductal capillaries and in the cytoplasm of some fibroblasts in the interstitial connective tissue. These data suggest that DPP IV in the submandibular and parotid glands may play some role in the secretion or reabsorption processes of secretory proteins and peptides in these glands.
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  • 34
    ISSN: 1432-0878
    Keywords: S-100 protein ; Thymus ; Interdigitating cells ; Immunocytochemistry ; Man
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    Notes: Summary The present study deals with the localization and ultrastructure of S-100-immunoreactive cells in the human thymus. These immunoreactive cells are distributed mainly in the medulla with some scattered elements in the cortex. Electron-microscopic observation revealed that the cells are characterized by an irregularly shaped nucleus, tubulovesicular structures in the cytoplasm and characteristic interdigitations of the plasma membrane. The cells often embrace lymphocytes with their branched processes. On the basis of these morphological features, the immunostained elements were identified as interdigitating cells (IDCs). The immunocytochemistry for S-100 visualizes the precise distribution and extension of the IDCs under the light microscope and indicates that the IDCs form no structural networks such as those established by the thymic epithelial cells. Since the IDCs in human lymph nodes have also been reported to contain S-100-like immunoreactivity, S-100 protein can be regarded as a useful marker for identifying the IDCs in the human thymus and other lymphoid organs.
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  • 35
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    Keywords: Immunocytochemistry ; FMRF-amide ; Neurotransmitter ; Palaemon
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary By use of a specific antiserum against the molluscan cardio-excitatory tetrapeptide FMRF-amide in combination with the PAP-method it was possible to obtain positive immunocytochemical reactions in several neurosecretory regions of the eyestalk of the prawn Palaemon serratus. FMRF-amide-like material was found in perikarya and nerve fibers of the medulla terminalis and in neurons in the lamina ganglionaris. The immunoreactivity observed in the glandular tissue located at the basal insertion of the eyestalk muscles must be ascribed to a non-specific reaction. The identification of immunopositive nerve fibers, ending on a nerve bundle in the medulla terminalis, and the fact that immunoreactive material was absent in the neurohemal sinus gland seem to indicate a neurotransmitter/neuromodulator function.
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  • 36
    ISSN: 1432-0878
    Keywords: Fetal porcine pituitary ; ACTH, MSH, β-LPH, α- and β-endorphin, GH, PRL ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The aim of the present study on the fetal porcine pituitary was (1) to detect by means of the immunoperoxidase technique the earliest stages of cells producing polypeptide hormones: β-MSH, ACTH, β-LPH, α- and β-endorphin, growth hormone (GH) and prolactin (PRL), (2) to study the development of the synthesis and the storage of these hormones during fetal life, and (3) to detect whether several hormones can be located in one and the same cell. The corticotropic cells were revealed as the earliest functional elements of the fetal anterior pituitary. Our results indicate clearly that ACTH, β-MSH, β-LPH, α- and β-endorphin appear at 34 days in the same regular, round or ovoid cells; no differences in the time of their appearance could be observed. The ACTH-cells, irregular or angular in shape and endowed with cytoplasmic processes such as described in the adult pituitary, were not seen until day 50. The first GH-cells were detected between 40 to 45 days of fetal life. From day 45 to 90, the GH-cells greatly increased in number and in staining intensity of their progressively extending cytoplasmic area, but they displayed the same regular and round shape. The PRL-cells were the last cell type to appear in the fetal pituitary. The first PRL-cells, small in size and round or ovoid in shape with a high nucleus/cytoplasm ratio, were detected at day 70. At day 80, the PRL-cells increased in size and staining intensity. They displayed an irregular elongated or stellated shape and cytoplasmic processes resembling those characteristic of the adult pituitary. These data suggest that in the fetal porcine pituitary: (1) ACTH, β-LPH and related peptides are synthesized and stored in the same cells, and (2) PRL and GH appear in individual cellular elements.
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  • 37
    ISSN: 1432-0878
    Keywords: Anterior pituitary, porcine ; Gonadotropic hormones (FSH, LH) ; Immunocytochemistry ; Cellular compartments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pre- and post-embedding immunocytochemical methods based on the use of specific antibodies against β-subunits of porcine LH and FSH were applied to determine the changes occurring in the anterior pituitary of the pig after gonadectomy. The results showed that (1) the total number of immunoreactive gonadotropes increased from 21–25% in control animals to 24–37% in castrated animals; (2) all gonadotropes contained both LH and FSH; (3) several types of immunoreactive LH/FSH cells were revealed; and (4) the two immunocytochemical methods used with dispersed cells localized the hormones in the same subcellular sites. However, the staining intensity in the different locations varied depending on the method applied. With the post-embedding method, a dense reaction product was found in the secretory granules but the cisternae of RER and the Golgi saccules were always slightly reactive. After the pre-embedding method, the staining intensity in the RER-cisternae and in the Golgi saccules was greatly increased. Thus, the two methodological approaches used in this study have permitted to visualize immunocytochemically the gonadotropic hormones not only at the sites of their storage but also along the intracellular pathway of the secretory material, i.e., at the site of its synthesis and during its passage via the Golgi zone.
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  • 38
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    Cell & tissue research 236 (1984), S. 171-180 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ultrastructure ; Supraoptic nucleus ; Neuropil ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neuropil located ventral to the SON was investigated by the use of immunoperoxidase staining for neurophysins, oxytocin and vasopressin, and electron miroscopy. The study was performed in six groups of rats: 1) control; 2) infusion of isotonic saline into the CSF; 3) infusion of hypertonic saline into the CSF; 4) drinking hypertonic saline for 4 days; 5) same as group 4 but injection of colchicine into the CSF on second day of dehydration; 6) salt loading for 3 months. In the control rats the ventral neuropil contained a few immunoreactive processes, the general morphology of which was completely different from that of the neurosecretory axons emerging from the SON at its dorsal aspect. In rats of groups 3 to 6 the ventral processes (VP) became loaded with neurosecretory granules, whereas the perikarya and axons were depleted. Based on their general morphology and reactivity pattern it is suggested that the VP are dendrites. Most of these “dendrites” were embedded in a glial cushion formed by the processes of a particular type of marginal glia. Some of these “dendrites” enveloped an arteriole penetrating the optic tract. All VP were rich in synaptic contacts. The possibility that the VP of neurosecretory cells may be functionally related to the subarachnoid CSF and the arteriolar blood flow is discussed.
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  • 39
    ISSN: 1432-0878
    Keywords: ACTH cells ; Immunocytochemistry ; Morphometry ; Spontaneous hypertension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary ACTH-immunoreactive cells in the anterior pituitary of 4-week-old spontaneously hypertensive rats (SHR) were studied with immunocytochemical and morphometric techniques. The results were compared with data from age-matched normotensive Wistar-Kyoto rats (WKY). No significant differences were found in volume density and average size of ACTH-immunoreactive cells between these two strains. However, SHR showed a significantly larger anterior lobe (2 P 〈 0.01) than WKY, indicating that the total number of ACTH-immunoreactive cells in the anterior pituitary is greater in SHR than in WKY. These data are in agreement with radioimmunological determinations showing a significantly elevated content (2 P 〈 0.01) but only a moderately higher concentration (0.05 〈 2 P 〈 0.10) of ACTH in the anterior pituitary of SHR as compared to WKY. The present results suggest an enhanced availability of ACTH in the anterior pituitary of 4-week-old SHR, a fact which could explain the markedly enhanced stress-induced release of ACTH previously found in these animals. This study further supports the hypothesis that, among other factors, an instability of the hypothalamo-pituitary-adrenal axis may contribute to the development of genetically programmed hypertension.
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  • 40
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    Cell & tissue research 236 (1984), S. 517-525 
    ISSN: 1432-0878
    Keywords: Bone marrow ; Actin ; Cell motility ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Blood cells proliferate extravascularly in the bone marrow and enter the circulation by migrating through endothelial cells of venous blood sinuses. This migration, or diapedesis, was suspected to involve actin. To test for the presence and distribution of actin, sections of rat bone marrow were examined by indirect immunocytochemistry. Affinity purified rabbit antichicken gizzard actin antibody, and goat-antirabbit IgG-FITC, or goat antirabbit IgG colloidal gold probes were used. The migrating cell contacts the endothelial cell and forms a podosome (a cortical bleb). Immunocytochemistry shows this region to contain actin. As diapedesis proceeds the podosome deforms, then breaches the endothelial cell. At this time the anterior portion of the leukocyte shows heavy labeling for actin. When the migratory cell traverses approximately half of its length through the endothelial cell, actin appears prominent in the caudal region of the cell. The immunocytochemical data suggest that actin is nonrandomly distributed in leukocytes undergoing diapedesis and may be a component of the force-generating mechanism responsible for this transcellular migratory event.
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  • 41
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Goat ; Folliculo-stellate cell ; GH cell ; S-100 protein ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Goat pituitary glands were immunohistochemically studied with antisera for bovine S-100 protein, rat LHβ, FSH, TSHβ, prolactin, ovine GH, and porcine ACTH1–39 by use of the superimposition technique on adjacent sections. Folliculo-stellate (F-S) cells were divided into two categories on the basis of ultrastructural properties: One consisted of a mass of agranular cells in which the pseudolumina were equipped with microvilli and cilia. Elongate gap junctions were often observed among these cells. The other was a group of granulated cells with or without pseudolumina. In this group the gap junctions were shown to be disintegrated. The dense granules 150–250 nm in diameter began to accumulate in the cells. However, neither type of these F-S cells was immunostained for S-100 protein. On the other hand, numerous polygonal, elongate, irregular or stellate cells containing S-100 protein were distributed throughout the gland. Most of them were immunohistochemically identical with the GH cells laden with the secretory granules 250–450 nm in diameter, but some of them were identical to TSH and prolactin cells which immunostained faintly for S-100 protein. This appears to be the first demonstration of GH cells intensely immunostained for S-100 protein.
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  • 42
    ISSN: 1432-0878
    Keywords: Hagfish ; Brain ; Pituitary ; FMRF-amide ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Paraffin sections of brain and pituitary of the hagfish Eptatretus burgeri were immunostained with an antiserum to FMRF-amide. Immunoreactivity was visible in a large number of neurons in the posterior part of the ventromedial hypothalamus and in long neuronal processes extending cranially from the hypothalamus to the olfactory system and caudally to the medulla oblongata. FMRF-amide-like immunoreactivity was also found in cells of the adenohypophysis. These observations suggest that the hagfish possesses a brain FMRF-amide-like transmitter system and pituitary cells containing FMRF-amide-like material. Antisera to ACTH, α-MSH and pancreatic polypeptide gave no immunoreaction in hagfish brain or pituitary.
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  • 43
    ISSN: 1432-0878
    Keywords: Crustacean hyperglycemic hormone ; Astacus leptodactylus ; Immunocytochemistry ; Quantitative electron microscopy ; Secretory cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical staining demonstrates striking differences in staining intensity among individual crustacean hyperglycemic hormone (CHH)-producing cells in the eyestalk of the crayfish Astacus leptodactylus. Based on these differences we arbitrarily subdivided the CHH-cells into three categories representing increasing immunoreactivity respectively: + cells, + + cells, and + + + cells. Electron microscopic investigations reveal that these differences in immunostaining are correlated with differences in the numerical density of the neurosecretory granules in the cytoplasm and that these may reflect differences in activity among the CHH-cells. Morphometric analyses at the light- and electron-microscopic levels indicate that the three distinguished categories of immunopositive cells represent different stages in the CHH-synthesizing process of the cells. The results of the present study demonstrate the application of the PAP-technique at the light-microscopic level as a method to obtain information pertaining to the dynamics of secretory activity of the CHH-cells.
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  • 44
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    Cell & tissue research 237 (1984), S. 321-327 
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Immunocytochemistry ; Respiratory tract ; Ear, middle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nerve fibres displaying neuropeptide-Y (NPY) immunoreactivity are abundantly distributed in the respiratory tract of cats, guinea-pigs, rats and mice. Fine beaded NPY fibres were seen in whole-mount spreads of the middle-ear mucosa. In the nasal mucosa and in the wall of the Eustachian tube NPY fibres were numerous around arteries and arterioles but sparse in the vicinity of veins; single fibres were found close to the acini of seromucous glands. In the tracheobronchial wall NPY fibres occurred in the proximity of blood vessels, in the subepithelial layer and in the smooth muscle. Surgical and chemical (6-hydroxydopamine treatment) sympathectomy resulted in disappearance of adrenergic and NPY-containing nerve fibres in the nasal mucosa. Sequential staining with antibodies against dopamine-β-hydroxylase (DBH) and NPY revealed that DBH and NPY occur in the same perivascular nerve fibres in the nasal mucosa. The distribution of NPY fibres in the respiratory tract suggests multiple functions of NPY, such as regulation of local blood flow, glandular secretion and smooth muscle activity.
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  • 45
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Biogenic amines ; Vasotocin ; Lymnaea stagnalis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Consecutive sections of certain neurons in the central ganglia of the pond snail Lymnaea stagnalis appear to be immunoreactive to anti-dopamine and anti-serotonin. The Cerebral Giant Neurons stain in addition with antivasotocin. The observations indicate the presence of two biogenic amines within the same neuron and in addition their co-existence with a biologically active peptide.
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  • 46
    ISSN: 1432-0878
    Keywords: Retinal photoreceptors ; Opsin ; Optic nerve ; Immunocytochemistry ; Cyclostome, Myxine glutinosa
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    Topics: Biology , Medicine
    Notes: Summary Opsin-immunoreactive sites in the eye and optic nerve of the hagfish, Myxine glutinosa, were studied by use of light-microscopic pre- and postembedding peroxidase-antiperoxidase or avidin-biotin-peroxidase techniques, and the immuno-electron-microscopic protein A-gold method. At the light-microscopic level, a strong opsin immuno-reaction was obtained on the outer segments of the photoreceptor cells with sheep and rat antibodies against bovine (rhod)opsin. These outer segments were located in the marginal photoreceptor space and in follicles of the retina, as well as in the tubular lumen of the optic nerve. Ultrastructurally, two classes of outer segments can be distinguished; most of them exhibited a strong antiopsin reaction, while certain elements lacked immunoreactivity with the antisera employed. The protein A-gold particles marked opsin-immunoreactive sites on the photoreceptor membranes. The presence of opsin-immunoreactive material in the retina and optic nerve of the hagfish strengthens the view that this primitive eye lacking a cornea, lens and vitreous body is engaged in light perception. The morphological similarity between the eye and pineal tissue is discussed in connection with the absence of a pineal organ in this species.
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  • 47
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    Cell & tissue research 238 (1984), S. 113-119 
    ISSN: 1432-0878
    Keywords: Crustacea ; Gastrointestinal hormones ; Neuropeptides ; Immunocytochemistry ; Cuticle ; Cancer magister
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A gastric peptide from the Dungeness crab (Cancer magister), extracted and characterized previously (Larson and Vigna 1983b), was localized in the foregut (stomach) of this species by immunocytochemistry using antisera specific for the bioactive carboxy-terminal amino acid sequence common to gastrins and cholecystokinins (CCKs). Immunoreactivity was found in all gastric epithelial cells and in the procuticle. Electron microscopy revealed an absence of peptidergic secretory granules in the gastric epithelial cells. The pattern of immunostaining suggests that the gastric epithelial cells secrete this peptide apically where it is incorporated into the cuticle lining the lumen. Specific immunostaining could not be demonstrated in various neural ganglia or in the hypodermis. The distribution of this peptide is different from that of gastrin/CCK in vertebrates and other invertebrates. This suggests that the crab gastric peptide is sufficiently similar to gastrin/CCK to react with C-terminal specific antisera, but may be anatomically, functionally, and possibly phylogenetically otherwise unrelated.
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  • 48
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    Cell & tissue research 238 (1984), S. 289-296 
    ISSN: 1432-0878
    Keywords: Somatostatin (SRIF) ; Brain ; Pituitary gland ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical investigations show that somatostatin (SRIF)-like immunoreactive material is present in the brain and the pituitary of nine different species of teleosts. In the brain, immunoreactive perikarya and fibers are observed in the preoptic periventricular nucleus, the entopeduncular nucleus, the anterior periventricular nucleus, and the nucleus lateralis tuberis. In the pituitary, SRIF-like-immunoreactive fibers occur in the proximal pars distalis (PPD), which contains the growth hormone (GH)-secreting cells. Nerve fibers are scattered among GH cells (cyprinids), or end on the basal lamina at the neuroglandular interface of the PPD (eel, salmonids). In the eel, the proximal neurohypophysis does not penetrate deeply into the PPD that is very poorly vascularized. In some species, e.g. Myoxocephalus, SRIF-like immunoreactive fibers are also observed in the caudal neurohypophysis, and even among MSH cells of the pars intermedia. In long-term starved carps and eels, the amount of SRIF-like material in the pituitary is clearly reduced. A possible role of SRIF in the concomitant stimulation of GH cells is discussed.
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  • 49
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Slug brain ; Somatostatin-like material ; Maturation hormone
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    Topics: Biology , Medicine
    Notes: Summary Immunocytological tests reveal the presence of a somatostatin-like substance in perikarya and axons in the brain of the giant slug Limax maximus L. Controls carried out on adjacent sections with absorbed antiserum or different antibodies raised against several biologically active peptides of vertebrates (ACTH-17-39, α- and β endorphin, α- and β MSH, methionin-enkephalin, TRH) demonstrate the specificity of the “staining”. However, some cells are both somatostatin- and FMRF-amide-positive. In the cerebral ganglia, the right Z-area cells, responsible for the synthesis of the maturation hormone (MH) are strongly somatostatin-positive. These results suggest a similarity between the MH and the somatostatin-like material contained in the Z-area cells. The simultaneous presence of two peptides in one and the same cell, the nature (elementary granules or soluble product) of the material, and its site of release are discussed.
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  • 50
    ISSN: 1432-0878
    Keywords: FMRF-amide ; Immunocytochemistry ; Nervous system ; Microstomum lineare (Turbellaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The central nervous system (CNS) and the peripheral nervous system (PNS) of the flatworm Microstomum lineare were studied by means of the peroxidase-antiperoxidase (PAP) immunocytochemical method, with the use of antisera to the molluscan cardioactive peptide FMRF-amide. FMRF-amide immunoreactive perikarya and nerve fibres are observed in the CNS and the PNS. In the CNS, immunoreactive perikarya and nerve fibres occur in the brain, in the epithelial lining and the mesenchymal surroundings of the ciliated pits, and positive fibres in the longitudinal nerve cords. In the PNS, immunoreactive fibre bundles with variocosities occur in the pharyngeal nerve ring, in symmetrical groups of perikarya on each side of the pharynx, and in the mouth area. Positive perikarya and meandering nerve fibres appear in the intestinal wall. A few immunoreactive cells and short nerve processes are observed at the male copulatory organ and on both sides of the vagina. Some immunoreactive peptidergic cells do not correspond to cells previously identified by histological techniques for neurosecretory cells. The distribution of immunoreactivity suggests that the FMRF-amide-like substance in CNS and PNS in this worm has roles similar to those of the brain-gut peptides in vertebrates. The status of FMRF-amide-like peptides as representatives of an evolutionarily old family of peptides is confirmed by the positive immunoreaction to anti-FMRF-amide in this primitive microturbellarian.
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  • 51
    ISSN: 1615-6102
    Keywords: Infection ; Invasion ; Multicellular Root Hairs ; Parasponia ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The first of two major steps in the infection process in roots ofParasponia rigida (Ulmaceae) following inoculation byRhizobium strain RP501 involves the invasion ofRhizobium into the intercellular space system of the root cortex. The earliest sign of root nodule initiation is the presence of clumps of multicellular root hairs (MCRH), a response apparently unique amongRhizobium-root associations. At the same time or shortly after MCRH are first visible, cell divisions are initiated in the outer root cortex of the host plant, always subjacent to the MCRH. No infection threads were observed in root hairs or cortical cells in early stages. Rhizobial entry through the epidermis and into the root cortex was shown to occur via intercellular invasion at the bases of MCRH. The second major step in the infection process is the actual infectionper se of host cells by the rhizobia and formation of typical intracellular infection threads with host cell accommodation. This infection step is probably the beginning of the truly symbiotic relationship in these nodules. Rhizobial invasion and infection are accompanied by host cortical cell divisions which result in a callus-like mass of cortical cells. In addition to infection thread formation in some of these host cortical cells, another type of rhizobial proliferation was observed in which large accumulations of rhizobia in intercellular spaces are associated with host cell wall distortion, deposition of electron-dense material in the walls, and occasional deleterious effects on host cell cytoplasm.
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  • 52
    ISSN: 1615-6102
    Keywords: Bauhinia purpurea ; Colloidial gold ; Golgi apparatus ; Immunocytochemistry ; Lectin ; Lowicryl K4M ; Protein body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The seed lectin of the tree legume,Bauhinia purpurea alba, was localized by electron microscopic immunocytochemistry. The pattern of lectin deposition and site of intracellular localization was examined in mid- to late-maturation seeds. The seed tissue was embedded in Lowicryl K4M, the use of which with seed tissues is discussed. Immunocytochemical labeling was accomplished with colloidal gold coupled to a second antibody. The immunocytochemical reaction was specific and sensitive. Protein bodies, Golgi apparatus and Golgi secretion vesicles were densely labeled. Golgi apparatus was oriented such that Golgi secretion vesicles were in close proximity to the protein bodies. The entire Golgi apparatus was labeled with no concentration gradient across the Golgi stack. These observations indicate that the final site of lectin deposition is the protein body, and that the Golgi apparatus plays an essential role in the deposition process.
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  • 53
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    Protoplasma 122 (1984), S. 86-90 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Non-articulated laticifers ; Articulated laticifers ; Asclepiadaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary sensitive immunocytochemical method for the identification of laticifers has been developed. Frozen sections of various laticifer-bearing plant material, mounted on slides, were first flooded with the IgG fraction of rabbit anti-latex antiserum, prepared using whole latex ofAsclepias syriaca, then flooded with fluorescein-conjugated IgG fraction goat anti-rabbit IgG to visualize laticifers. Positive fluorescence was observed for laticifers in shoots and embryos ofA. syriaca andStapelia bella and embryos ofA. tuberosa. Laticifers did not fluoresce in shoots ofA. tuberosa andEuphorbia tirucalli, in embryos ofE. marginata, or in petioles ofMusa paradisiaca andCichorium intybus. Controls prepared with uninjected rabbit serum were negative (no fluorescence).
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  • 54
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    Protoplasma 121 (1984), S. 25-33 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Colloidal gold ; Wheat germ agglutinin ; Lectin ; Cryosections
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The chitin-binding lectin wheat germ agglutinin (WGA) is found at the periphery of wheat embryos, and a similar lectin is present at the root tips of older plants (Mishkind et al. 1982). Although a ferritin-conjugated secondary antibody is adequate for localizing WGA in embryos, native electron-opaque particles make the electron microscope identification of added label equivocal in other wheat tissues. As reported here, however, unambiguous ultrastructural localization of WGA-like lectin in adult wheat roots can be obtained with rabbit anti-WGA followed by colloidal gold-labeled goat anti-rabbit (GAR) IgG. Colloidal gold (CG) was prepared by the reduction of gold chloride with citrate, ascorbate or phosphorous. GAR IgG, prepared from serum by antigen affinity chromatograhy, was adsorbed to the gold particles to produce a stabilized suspension of GAR-CG. Localization was performed on 8–12 μM frozen sections of tissue fixed in 4% paraformaldehyde, 0.3% glutaraldehyde, and 0.75% acrolein in phosphate-buffered saline containing 1M sucrose. Localization with GAR-CG was first compared to that ascertained in embryos using other probes and was then extended to the roots of adult plants. An advantage of the GARCG method is that it permits the visualization of antigen at both the light and electron microscope levels in the same section. At the light level, the anti-WGA-GAR-CG complex appears as a red stain that is localized in specific tissues of embryos and in the caps and outer layers of adult roots. Sections in which lectin was detected at the light microscope level were embedded in plastic and sectioned for subcellular examination. Electron dense gold particles indicative of WGA are found at the periphery of protein bodies in wheat embryos and in vacuoles of the roots of adult plants. Sections incubated with control IgG lack reaction product.
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  • 55
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    Plant and soil 76 (1984), S. 127-137 
    ISSN: 1573-5036
    Keywords: Adenylate pool ; Biomass volume ; CO2 evolution ; Chitin ; DNA ; Electron microscopy ; Enzymes ; Fluorescent antibody ; Fumigation-respiration ; Fungi Histochemistry ; Imunofluorecence ; Jones-Mollison technique ; Microcosms ; Monoclonal antibodies ; Nitrogen ; Nutrients ; Oxygen consumption ; Phosphorus ; Phytotoxins ; Plate counts ; Rhizobium ; Rhizosphere ; Sulphur ; Xenobiotics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary There is an immense literature on biological and biochemical analyses of soils. Such analyses have revealed the enormous richness of species in soil and their vast range of metabolic potentials and ecological diversity. Accordingly, the approaches used to investigate the soil biota and its biochemistry usually have to be modified or adapted depending upon the purpose of the investigation. Studies of micro-organisms in the soil environment, are complicated because microbial cells are commonly attached to surfaces where they live side-by-side with other populations in consortia usually containing different morphological and physiological types. Such assemblages of organisms cannot be described quantitatively using cultural techniques, such as plate counts, which underestimate both cell numbers and viable biomass. The development of more powerful observational and staining techniques has improved our knowledge of the diverse morphological and biochemical composition of soil micro-communities. Such findings have been amplified at a grosser level by laboratory studies with multi-component systems (microcosms) to mimic field situations and to assess the range of biochemical potentials of microbial consortia. But despite notable advances in analytical methods we are still, with a few exceptions, unable to detect or identify those microorganisms which carry out specific biochemical transformations or determine whether particular cells are alive, dormant or dead at the time of observation. Considerable work has been done to define some of the fundamental ecological attributes of microbial assemblages in soil. Productive work on the metabolic activities of the soil microbiota, specially geochemical transformations of C, N, S and P, has been under way for more than a century. But only in more recent years have more sensitive and reproducible analytical methods become available to measure viable biomass in soil. This will enable some insight to be gained into the role that microbial biomass plays as a labile source and sink for plant nutrients.
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  • 56
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    Plant and soil 77 (1984), S. 387-390 
    ISSN: 1573-5036
    Keywords: Antibiosis ; Culture filtrate ; Fungi ; Peanut ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Four strains ofRhizobium sp. from peanut (Arachis hypogaea L.) root nodules were tested for their sensitivity to metabolites (culture filtrates) of more than ten common soil inhabiting fungi, in yeast extract mannitol agar and broth cultures. Among the rhizobial strains tested strain CB-530, BU-1 and BU-2 were not sensitive to metabolites ofMyrothecium roridum andFusarium moniliforme whereas CB-1024 was sensitive. Culture filtrates ofGliocladium roseum, Thielavia basicola andDrechslera pedicellata inhibited the strains CB-530 and BU-2 but not CB-1024. Most of the soil inhabiting fungi tested were inhibitory to rhizobial growthin vitro and very few were stimulatory, their effect in either case being strain specific.
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  • 57
    ISSN: 1573-5036
    Keywords: Acid soils ; Inoculant ; Rhizobium ; Soybeans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Rhizobial inoculation trials were conducted in an acid heavy clay soil in Mekong Delta, Viet Nam, using peat based inoculants produced locally and the commercial granular product of Nitragin CCo., Wisconsin, USA. The pH of these soils ranged from 4.5 to 5.1. Two soybean cultivars, MTD6 and MTD10, were tested as host plants. There were no significant differences between locally made inoculant treated plants and the uninoculated controls in both cultivars. But, the Nitragin inoculation improved all plant characteristics examined in both cultivars. Grain yields of Nitragin inoculated plants of cultivar MTD6 and cultivar MTD10 were 6.5 and 5.5 times as much as those of the controls; protein content of grain increased 11 and 16 percent, respectively. Well nodulated plants had shorter life cycles, flowering durations, and days to flowering. The Rhizobium symbiosis resulted in an additional 153 kg grain-N/ha. These studies show that a surface coated commercial multistrain inoculant can be used to successfully grow soybeans in the acid, heavy clay soils of the Mekong Delta.
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  • 58
    ISSN: 1573-5036
    Keywords: Acid soils ; Nitrogen fertilizer ; Rhizobium ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Soybean cultivar MTD10 cultivated on a moderate acid paddy soil of the Mekong Delta responded well to a multistrain inoculation and to chemical nitrogen fertilizer. However, the efficiency of chemical nitrogen uptake by the uninoculated plants decreased with increasing nitrogen application rate. Inoculation improved all examined agronomic characteristics of plants. Inoculated plants produced grain yield 10 times greater than the uninoculated controls and 2.5 times greater than the uninoculated plants receiving 80 kg/ha of chemical nitrogen. Inoculation was superior to chemical nitrogen fertilization in all parameters of importance to market-oriented farmers.
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  • 59
    ISSN: 1573-5036
    Keywords: Hill soils ; Lime ; Mycorrhiza ; Nitrogen fixation ; Nodulation ; Phosphorus ; Rhizobium ; Symbioses ; White clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The responses of white clover (cv NZ Grasslands Huia grown in four UK hill soil types) to additions of lime and P, to inoculation with Rhizobium and mycorrhizal fungi, and to differences in soil water status were assessed in pot and field experiments. With a deep peat soil in pots, shoot production, nodulation and N fixation by clover were increased by 160, 130 and 85% respectively following inoculation with mycorrhiza, but in the field, despite a doubling of root infection, there was no response in growth. On a brown earth soil in the field inoculation with one endophyte (Glomus mosseae L1) out of four tested depressed production of white clover shoots by 42% but enhanced that of leeks (Allium porrum) by 50%; the others were without effect. With dry peaty podzol and brown earth soils in pots, clover shoot production was highest with added P when a water holding capacity of 80% was maintained, but roots from the latter had only 2.6 compared to 68 nodules per plant from the former. Further work is required to explain poor nodulation in the brown earth soils.
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  • 60
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    Plant and soil 78 (1984), S. 445-448 
    ISSN: 1573-5036
    Keywords: Leucaena ; Rhizobium ; VA mycorrhiza
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Response ofLeucanea leucocephala to inoculation withGlomus fasciculatum and/or Rhizobium was studied in a phosphorus deficient unsterile soil.G. fasciculatum only inoculation improved nodulation by native rhizobia and Rhizobium only treatment improved colonization of roots by native mycorrhizal fungi. Dual inoculation with both the organisms improved nodulation, mycorrhizal colonization, dry weight, nitrogen and phosphorus content of the plants compared to single inoculation with either organism.
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  • 61
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    Plant and soil 78 (1984), S. 381-391 
    ISSN: 1573-5036
    Keywords: Aluminium ; Caloium ; Nodulation ; pH Phosphate ; Polymeric hydrolysis ; Rhizobium ; Rhizosphere ; Root elongation ; Root hairs ; Trifolium repens ; White clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Effects of aluminium on theTrifolium repens var Huia-Rhizobium trifolii strain HP3 symbiosis were studied using an axenic solution-culture system. With, 10 μM phosphate, 50 μM aluminium reduced or inhibited root elongation at pH〈5.0, root hair formation at pH〈 5.0–5.5, and Rhizobium multiplication in the rhizosphere and nodule formation at pH〈6.0. In the absence of aluminium, root elongation and root hair formation were reduced at pH〈4.3, and Rhizobium multiplication and nodule formation were inhibited at pH〈5.0. Root hair formation was more sensitive to aluminium at pH〈5 than was root elongation. No effect of aluminium on Rhizobium multiplication and nodule formation at pH〈5 was detected because both were sensitive to pH alone. At pH 5.5 most of the aluminium changed immediately to a form which was susceptible to low-speed centrifugation, but which was detected by the aluminon method of analysis, and after 24 h a precipitate formed. the concentration of phosphate was reduced also, to approximately 1μM. Toxicity was overcome by either increasing the phosphate concentration from 10 to 50 μM, or by increasing the pH to 6.0 and the calcium, concentration to 1000μM.
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  • 62
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    Plant and soil 80 (1984), S. 407-415 
    ISSN: 1573-5036
    Keywords: Cellulase ; Infection process ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The production of cellulase byRhizobium species was studied.Rhizobium trifolii cellulase was induced by a variety of polysaccharides, including celluloses and hemicelluloses. Cellobiose and myo-inositol also allowed enzyme expression but mannitol prevented it at concentrations higher than 0.25%. Both soluble and insoluble plant root substances moderately stimulated cellulase production byRhizobium trifolii. Most substances tested did not induce the production of cellulases by the “slow-growing, cowpea type” rhizobia strain CIAT 79. Effective inducers were carboxymethylcellulose, gluconate and myo-inositol. Cellulase production was very low under all conditions tested. In most cases the enzyme activity was loosely bound to the capsular material. The enzyme in fast-growers is an 1,4-β-D-glucan-4-glucanohydrolase (endo-glucanase EC 3.2.1.4) with specificity for high molecular weight polysaccharides. There was no correlation between infectiveness ofRhizobium trifolii strains and cellulase production. One strain, which lacks the nodulation plasmid, produced cellulase at the same rate as its parental infective strain.
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  • 63
    ISSN: 1573-5036
    Keywords: Acidity ; Aluminium ; Cowpea ; Nitrogen ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The interaction of pH (4 or 6), aluminium (0 or 16 ppm at pH 4) and N source (symbiotic or combined) on the growth and nutrient status of cowpea (Vigna unguiculata) was studied in a glasshouse experiment. Low pH significantly decreased the growth of the plants dependent on symbiotic nitrogen fixation but at pH 4 the addition of 16 ppm Al further depressed growth in both nitrogen regimes. Al-ions appear to exert their effect primarily on the root system, as shown by the reduction in total length and fresh weight. The symbiotic development of the plants was affected by low pH but more markedly by the Al treatment. Shoot nitrogen concentrations were reduced from ca. 2.6% at pH 6 to 1.8% and 0.9% at pH 4 without and with aluminium respectively. Calcium concentration was decreased by low pH and further by Al in both nitrogen regimes. In all Al-treated plants, the aluminium was mainly accumulated in the roots and was associated with an increase in their phosphorus concentration.
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  • 64
    ISSN: 1573-5036
    Keywords: Glycine max ; Inoculation ; IW/CPE ratio ; Nitrogen accumulation ; Nitrogen harvest ; Nitrogen re-distribution ; Nodulation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A field experiment was conducted on soybean (Glycine max (L.) Merrill) with a view to find out the effect of seed inoculation and scheduling of irrigation on nodulation, accumulation and re-distribution of nitrogen in plant tops and soil. The eight treatment combinations consists of two seed inoculations,viz. uninoculated and inoculated with rhizobium culture, and four irrigation schedules,viz. irrigation water to the cumulative pan evaporation (IW/CPE) ratio of 0.5, 0.7, 0.9 and a control (rainfed). Seed inoculation by, rhizobium culture increased the number, dry-weight and N content of nodules per plant. Inoculation of seeds also increased the N accumulation rate in plant top and it was 2.48 kg/ha/day during the flower-initiation to the pod-initiation stage (30–60 days interval). At harvest, 32.2, 47.8 and 26.2 kg N/ha was re-distributed from the stems, leaves and pods-wall of inoculated plants to the grains, respectively. A total of 186.5 kg N/ha was harvested and 64.7 kg N/ha, was accumulated in soil under the inoculated condition. Scheduling of irrigation at 0.7 IW/CPE proved better, than other irrigation schedules and helped in increasing the nodulation, nitrogen accumulation and grain yield. As compared to control, 8.4, 17.8 and 18.4 kg more of N/ha was redistributed from the stems, leaves and pods-wall respectively when the irrigations were scheduled at 0.7 IW/CPE ratio. Under this irrigation schedule the total N harvest was 200.1 kg/ha while the total N increased by 55.9 kg over that present in soil at the time of sowing.
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  • 65
    ISSN: 1573-5036
    Keywords: Clover ; Nitrogen fixation ; Rhizobium ; Root nodule ; Selection and plant breeding ; Symbiosis ; Trifolium pratense ; T. subterraneum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary This paper reviews (i) basic studies on the genetics of symbiosis in red clover (a self-sterile species) and subterranean clover (cleistogamous) and (ii) work on selection and plant breeding to increase nitrogen fixation in these hosts. Symbiotic effectiveness in red clover is influenced by many major and minor genes. The highly effective phenotype is inherited in a complex manner associated with early nodulation and the formation of large amounts of persistent bacteroid-containing tissue. Lines bred to fix more nitrogen with one strain ofRhizobium trifolii do so with most but not all other strains examined. They also show slightly increased vigour when grown on nitrate. The highly effective response is correlated with abundant nodulation and an early flowering habit, the evidence from breeding studies indicating that this correlation is not absolute. Normally effective and highly effective nodules have the same specific nitrogenase activities. The expression of the highly effective response is relatively little affected by environmental factors (temperature, light intensity, day length, supplementary carbon-di-oxide). Inbreeding substantially degrades the symbiotic response. Heterosis is shown in crosses between cultivars of subterranean clover but otherwise selection to increase effectiveness in this host was unsuccessful. The relevance of these results (and their physiological aspects) for the improvement of grain legumes is discussed.
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  • 66
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    Plant and soil 82 (1984), S. 377-386 
    ISSN: 1573-5036
    Keywords: Centrosema ; Desmodium ; Inoculation ; Nitrogen yield ; Nodulation ; Oxisol ; Pueraria ; Rhizobium ; Soil cores ; Stylosanthes ; Tropical forage legumes ; Zornia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Three phases of Rhizobium inoculation trials were carried out as part of a programme to select forage legume germplasm adapted to acid, infertile Oxisols of tropical America. Firstly, a range of tropical forage legumes were evaluated for their response to N fertilization or inoculation with strains previously shown to be effective in Leonard jars, using cores of undisturbed soil or in the field at Carimagua, Meta, Colombia. In pure legume stands onlyCentrosema macrocarpum andC. pubescens showed increases in N yield due to both inoculation and N fertilization;C. brasilianum responded only to N fertilization;Zornia latifolia, Z. brasiliensis andStylosanthes capitata responded to neither treatment. Trials in cores and in grass-legume mixtures showed responses ofDesmodium ovalifolium, Pueraria phaseoloides andS. capitata to N fertilization but not to inoculation. In the second phase of experiments strains were screened in soil cores with 16 ecotypes ofDesmodium, Centrosema, Stylosanthes andPueraria spp. Significant increases in N yield due to inoculation occurred with at least one strain in all the legumes exceptS. guianensis ‘tardio’, and in some trials withS. capitata. In the third phase of trials the most effective strains were tested in the field. Significant response ofP. phaseoloides andC. macrocarpum to inoculation at two sites and in the second year after establishment were shown. Further screening trials and field trials at different sites are needed in order to provide better recommendations for inoculation of grazing trials being set up in the region under study.
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  • 67
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    Plant and soil 82 (1984), S. 273-284 
    ISSN: 1573-5036
    Keywords: Legume ; Nitrogen fixation ; Rhizobium ; C and N economy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Studies of the C and N economy of a range of temperate and tropical legume/Rhizobium symbioses indicate considerable variation (up to three-fold) in the cost of N2 fixation. Comparisons between and within symbioses indicate that the proportion of net photosynthate utilized in nodule functioning varies almost ten-fold from as low as 3% to as high as 25%. Factors possibly responsible for variation in efficiency of C use in nodules and in the proportioning of translocated photosynthetic products to nodules are discussed.
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  • 68
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    Plant and soil 82 (1984), S. 329-335 
    ISSN: 1573-5036
    Keywords: Legume breeding ; Medicago sativa ; Nitrogen fixation ; Rhizobium ; Trifolium repens ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary This paper examines evidence which quantifies the relative importance of legume and Rhizobium genotypes as determinants of phenotypic variation in symbiotic nitrogen fixation. It demonstrates potentially large and unpredictable effects of the Rhizobium genotype. The likely importance of such effects on crop yield is considered. The information is then used to assess ways in which legume breeding programmes may be altered to encompass the effects of genetic variation in Rhizobium.
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  • 69
    ISSN: 1573-5036
    Keywords: Abscisic acid ; Gibberellin ; Polyethylene glycol ; Rhizobium ; Roots ; Nitrogenase ; Nodulation ; Soya ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effect of 2-day cycles of osmotically induced leaf moisture stress followed by partial recovery on the nodulation and nitrogenase activity of 2 soya cultivars was studied. Fourteen days after plant inoculation (mid-growth stage) the total leaf electrochemical water potential (ψwleaf) of control plants ranged from −0.8 to −1.9 bars, whereas the concentrations of osmoticum (polyethylene glycol 4000) induced ψwleaf values ranging from −1.4 (recovery value) to −3.1 bars (low stress), −1.8 to −4.4 bars (mild stress), and −2.2 to −6.2 bars (medium stress). The low stress treatment reduced nodule numbers and their specific activity in both cultivars, without affecting nodule size or the time required for nodule initiation. Nodule initiation was delayed in both cultivars by the mild and medium stress treatments, the former treatment reducing the number and size of the nodules and such nodules exhibited very low specific activity. The medium stress treatment prevented the further development of nodule initials, which remained inactive throughout the experiment. Such results imply an effect of water stress on the infection process and on nodule morphogenesis. The reduction in nodule numbers observed in water stressed plants was not associated with a reduced number of rhizobia in the rhizoplane nor due to an effect on root growth or root hair formation. At a stage prior to the formation of macroscopic nodule initials, the roots of plants under medium stress (ψwleaf=−5.5 bar)s) had a higher content of abscisic acid (ABA) (4-fold increase) and a lower content of gibberellin (GA)-like substances (21.4% reduction) as compared to control plants (ψwleaf=−1.0 bar). Although the medium stress treatment slightly increased the stomatal resistance of leaves, photosynthetic and transpiration rates were unaffected. Similar alterations of the hormononal balance occurred in the nodulated roots of plants subjected to naturally induced leaf moisture stress. Since the foliar application of ABA (1.92×10−5 M) to unstressed plants inhibited nodulation (45% reduction in nodule numbers), the increased endogenous content of thishormone in the roots of plants under leaf moisture stress may provide some physiological insight into the inhibitory effect of water stress on the nodulation process.
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  • 70
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    Plant and soil 80 (1984), S. 297-300 
    ISSN: 1573-5036
    Keywords: Nodulation ; Non-nodulating soybean ; Rhizobitoxine ; Rhizobium ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A significant increase in nodulation of “non-nodulating” (rj1 rj1) soybeans was obtained by inoculating with very high numbers (approx. 1011 cells/pot) of certain rhizobia when compared with inoculation at a moderate dose (approx. 109 cells/pot). Nodulating ability of rhizobial strains was not correlated with their ability to produce a detectable level of chlorosis-inducing toxin in culture.
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