Xylem Sap from Actinidia chinensis: seasonal Changes in Composition

Seasonal variation was followed in the content of total a-aminoacids, arginine, calcium, total carbohydrate, ß-galactosidaseactivity, magnesium, nitrate, phosphatase activity, phosphateand sulphate in vacuum-extracted xylem sap from Actinidia chinensisvar. hispida, the Chinese gooseberry or kiwifruit. There wasa marked increase in the concentration of most sap componentsjust prior to leaf emergence followed by a rapid decrease afterthe leaves had expanded. Experiments with excised extensionshoots showed that much of this spring-time increase in concentrationsof sap components was due to mobilization of nutrients withinthe shoot itself. Sap from the trunk and the older brancheschanged less in composition than did sap from the younger partsof the plant. The amplitude and direction of trends in concentrationof sap from the different parts of the plant varied with nutrientand with time. Analysis of vacuum-extracted xylem sap collectedduring periods of rapid transpiration from early summer onwardsgives a reliable indication of the composition of the transpirationstream. Actinidia chinensis, Chinese gooseberry, kiwifruit, storage reserves, xylem sap, amino acids, arginine, calcium, carbohydrate, ßgalactosidase, magnesium, nitrate, phosphatase, phosphate, potassium, sulphate     

Supply and uptake of potassium,calcium and magnesium of spray carnations (Dianthus caryophyllus) grown in rockwool

Summary In two experiments with carnations grown in rockwool the effects of different cation ratios in the nutrient solution were studied. The results showed that carnations need a high calcium supply. The crop did not appear to be sensitive to different potassium-magnesium ratios in the nutrient solution. In the nutrient solution added mole ratios K∶Ca∶Mg=55∶35∶10 seemed to be optimal. Such ratios in addition led to ratios of 55∶30∶15 in the root environment. Tissue analysis showed that in younger leaves of peduncles harvested a potassium content of 900 mmol per kg dry matter was optimal. For calcium a content of 350 and for magnesium 100–150 mmol per kg was needed. Analytical data of plant-sap analyses were closely correlated with data gained by digestion of dried material. For potassium and magnesium the relationships were linear. However, for calcium a curvilinear relationship was found. In the experiments indications were obtained that a sufficient calcium supply suppressed foot rot in carnations.     

Effects of Vitamin E and Selenium on Serum Trace and Major Elements in Horses

The combined effects of vitamin E and selenium were studied in native Anatolian horses subject to strenuous exercise. The concentrations of copper, zinc, iron, calcium, potassium, and magnesium were determined in serum by atomic absorption spectrometry in two study groups (n = 25 each), one of which served as untreated controls. After exercising the horses by running 1,500 m in about 7 min, only the copper level and the copper/zinc ratio significantly increased (p < 0.05), but the concentrations of calcium, potassium, iron, and magnesium remained unchanged. In horses treated with vitamin E and selenium, the calcium and potassium levels decreased to levels lower than those of untreated controls before and after exercise. The iron levels were not changed by exercise or treatment alone but increased when the horses had been supplemented and exercised. The copper level and the copper/zinc ration increased as a result of exercise in both treated and untreated horses. These changes suggest that supplementation with vitamin E and selenium had an important effect on the serum concentrations of calcium, potassium, copper, iron, and the copper/zinc ratio.     

Effect of Sodium, Potassium, Calcium, Magnesium and Tetraethylammonium on the Transient Voltage Response to a Galvanostatic Step and of the Temperature on the Steady Membrane Conductance of Chara corallina: a Further Evidence for the Involvement of Potassium Channels in the Fast Time Variant Conductance

Homble F. 1985. Effect of sodium, potassium, calcium, magnesiumand tetraethylammonium on the transient voltage response toa galvanostatic step and of the temperature on the steady membraneconductance of Chara corallina: A further evidence for the involvementof potassium in the fast time variant conductance.—J.exp. Bot. 36: 1603–1611. Potassium channels of Chara corallina have an activation energyof 36±1 kJ mol^–1 and 50±2 kJ mol^–1at temperatures higher and lower than 15°C respectively.The fast time variant conductance property of potassium channelsis insensitive to sodium and magnesium ions and is depressedby the presence of calcium, potassium and tetraethylammoniumions. It is suggested that in Chara two different kinds of potassiumchannels exist, each kind being distinguished by their kineticsand their response to calcium and magnesium ions. Key words: —Chara corallina, membrane conductance, potassium channels, temperature     

Redistribution of Phosphorus, Sulfur, Potassium and Calcium in Relation to Light-Induced Gravitropic Curvature in Zea Roots

Phosphorus, sulfur, potassium and calcium were measured by particle-inducedX-ray emission (PIXE) in horizontally oriented, light-irradiatedor non-irradiated primary roots of Zea mays L., cv. Golden CrossBantam 70 which exhibit gravitropic response only after exposureto light. The content of the four elements increased in thelower half of horizontally oriented roots which had been brieflyexposed to white or red light, while there were no marked differencesin distribution between the upper and lower halves of non-irradiatedroots. The increase of each element in the lower half was observed15–30 min after irradiation in root caps and 30–60min after irradiation in the elongation zones. The effect ofred light was not reversed by far-red light given immediatelyafter the red irradiation. Ethylene glycol-bis(ß-aminoethylether)-N, N, N',N'-tetraacetic acid (EGTA) treatment of roottips inhibited the gravitropic curvature of roots, and the additionof Ca reduced this inhibition. The meaning of Ca redistributionin root caps and elongation zones during light-induced gravitropiccurvature of maize roots is discussed. (Received December 4, 1985; Accepted March 22, 1986)     

Human melanoma and Chinese hamster ovary cells galactosylate n-alkyl-{beta}-glucosides using UDP gal:GlcNAc {beta}1,4 galactosyltransferase

We previously showed that human melanoma, CHO and other cellscan convert ß-xylosides into structural analogs ofganglioside G_M3. We have investigated several potential acceptorsincluding a series of n-alkyl-ß-D-glucosides (n =6–9). All were labeled with ³H-galactose when incubatedwith human melanoma cells. Octyl-ß-D-glucoside (GlcßOctyl)was the best acceptor, whereas neither octyl--D-glucoside norN-octanoyl-methylglucamine (MEGA 8) were labeled. Analysis ofthe products by a combination of chromatographic methods andspecific enzyme digestions showed that the acceptors first receiveda single Galß1,4 residue followed by an 2,3 linkedsialic acid. Synthesis of these products did not affect cellviability, adherence, protein biosynthesis, or incorporationof radio-labeled precursors into glycoprotein, glycolipid orproteoglycans. To determine which ß1,4 galactosyltransferase synthesized Galß1,4GlcßOctyl,we analyzed similar incubations using CHO cells and a mutantCHO line (CHO 761) which lacks GAG-core specific ß1,4galactosyltransferase. The mutant cells showed the same levelof incorporation as the control, eliminating this enzyme asa candidate. Thermal inactivation kinetics using melanoma cellmicrosomes and rat liver Golgi to galactosylate GlcßOctylshowed the same half-life as UDP-Gal:GlcNAc ß1,4 galactosyltransferase,whereas LacCer synthase was inactivated at a much faster rate.We show that GlcßOctyl is a substrate for purifiedbovine milk UDP-Gal:GlcNAc ß1,4 galactosyltransferaseFurthermore, the galactosylation of GlcßOctyl by CHOcell microsomes can be competitively inhibited by GlcNAc orGlcNAcßMU . These results indicate that UDP-Gal:GlcNAcß1,4 galactosyltransferase is the enzyme used forthe synthesis of the alkyl lactosides when cells or rat liverGolgi are incubated with alkyl ß glucosides. alkylglucosides galactosyltransferase glycolipid artificial acceptors     

Analysis of Trace and Major Elements in Bronchoalveolar Lavage Fluid of Mycoplasma Bronchopneumonia in Calves

The aim of this study was to evaluate the reliability and effectiveness of direct determination of trace and major element concentrations in bronchoalveolar lavage fluid samples from Holstein calves with Mycoplasma bronchopneumonia (n = 21) and healthy controls (n = 20). The samples were obtained during bronchoscopy using a standard examination method. A total of 18 elements (aluminum, bromine, calcium, chlorine, chromium, copper, iron, potassium, magnesium, manganese, molybdenum, nickel, phosphorous, sulfur, silicon, strontium, titanium, and zinc) were detected by particle-induced X-ray emission. The average bromine, iron, potassium, magnesium, and phosphorous concentrations were higher in calves with bronchopneumonia than in controls (p < 0.05). They were found to have higher amounts of calcium and zinc, and a higher zinc–copper ratio than that in healthy calves (p < 0.001). Based on the receiver operating characteristics curves, we propose a diagnostic cutoff point for zinc–copper ratio for identification of Mycoplasma pneumonia of 8.676. Our results indicate that assessment of the elemental composition of broncholaveolar lavage fluid is a promising diagnostic tool for Mycoplasma bronchopneumonia.     

The Interactions Between Monovalent Cations and Calcium During Their Adsorption on Isolated Cell Walls and Absorption by Intact Barley Roots

The adsorption of sodium, potassium, rubidium and calcium ionsat different concentrations was measured on cell walls frombarley roots (Hordeum vulgare L. cv. Union), prepared by detergenttreatment. It was found that sodium and calcium interacted verystrongly during their simultaneous adsorption, whereas potassiumdid not interfere with calcium. This has led us to concludethat calcium and sodium are adsorbed on identical sites in thecell wall, whereas potassium is adsorbed at another site. Rubidiumseems to be less specific for both sites and interferes onlymoderately with calcium. The adsorption on cell walls of thesecations was compared with their adsorption on intact roots at2 °C, where beside the cell wall, sites may be availableat the outer surface of the membrane, and further measurementswere made of absorption at 25 °C. The fact that sodium interactswith calcium and potassium alters the ratio of K to Na in thecell wall compared to their concentrations in the medium. Thepreferential shift towards potassium when calcium is presentcould be very important for the rates of initial uptake in lowsalt barley roots, since the membrane is in contact with a differentproportion of K to Na in the cell wall from the one suppliedin the medium. Hordeum vulgare L., barley, absorption of cations, adsorption, calcium, sodium, potassium rubidium     

Desiccation and the Control of Expression of {beta}-Phaseolin in Transgenic Tobacco Seeds

Drying of seeds at certain stages prior to maturation, i.e.premature desiccation, will terminate synthetic events uniqueto development, for example, storage protein synthesis, andinitiate processes associated with germination. In this studywe have investigated the role of desiccation in the expressionof a storage protein gene, ß-phaseolin, to determineif such a developmentally-regulated gene remains sensitive todrying when controlled by a promoter that has no known sensitivityto this treatment. We compared, in transgenic tobacco seeds,the effects of maturation and premature drying on the expressionof a full ß-phaseolin gene, and ß-phaseolingenes driven by a cauliflower mosaic virus 35S promoter withor without an alfalfa mosaic virus (AMV) 5' untranslated leadersequence. The results indicate that the ß-phaseolinpromoter is directly down-regulated by desiccation during maturationand, although activated during the drying phase of a prematuredesiccation event, it is not active upon rehydration or imbibition.The 35S promoter is down-regulated also by both maturation dryingand premature desiccation but unlike the ß-phaseolinpromoter it is reactivated upon rehydration or imbibition. Key words: Desiccation, ß-phaseolin, gene regulation, Phoseolus vulgaris, seed development     

Calcium, Sulfhydryls and the Mitotic Apparatus

The in vivo mitotic apparatus (MA) of clam, worm and sea urchineggs may be augmented or dispersed by application of specificantimitotic agents. Glycols, which are antimitotic at concentrationsof 1–3% in sea water, cause a rapid massive increase inMA volume and retardation as seen in the polarizing microscope.Caffeine and dinitrophenol (DNP) cause a rapid disappearanceof the MA by shrinkage. Glycol effects can be balanced by DNPor caffeine if the agents are applied at the proper time andconcentration although normal cleavage does not ensue. Analysisof DNP and caffeine shrinkage suggests that they act indirectlyby causing release of calcium from intracellular stores, calciumcausing inactivation of polymerizable tubulin. DNP could causerelease of calcium either from mitochondria (if egg mitochondriahave a calcium uptake system) or by causing a decrease in ATPlevels which would inactivate calcium uptake systems such asthe Petzelt Ca⁺² ATP-ase or Kinoshita Ca⁺² uptake vesicles. Caffeine, while an inhibitor of the phosphodiesterase for cyclicAMP does not cause inhibition of the MA through the cyclic nucleotide.It is found that caffeine (and other methyl xanthines) causean inhibition of glutathione reductase activity in treated eggs.It is postulated that the calcium ATP-ases in the egg may becontrolled by reversible oxidation and reduction of their sulfhydrylsthus regulating calcium concentration in the cytosol. The possiblemodes of action of this system are discussed.     

{beta}-Naphthyl oligophosphates: Inhibitors of photophosphorylation and H+-ATPase of spinach chloroplasts

ß-Naphthyl di-, tri- or tetraphosphate inhibits photophosphorylationof spinach chloroplasts competitively with ADP, whereas ß-naphthylmonophosphate inhibits it competitively with Pi. The apparentKi of ß-naphthyl diphosphate for the ADP site was300 µM and that of ß-naphthyl monophosphatefor the Pi site was 1.45 mM. At 10 mM, both of these two organicphosphates inhibited photophosphorylation more than 90%. Noneof the above four ß-naphthyl phosphates were phosphorylatedby chloroplasts. ß-Naphthyl di-, tri- or tetraphosphateinhibits ATPase activity of isolated chloroplast coupling factor1 (CF₁) (EC 3.6.1.3 [EC] ) and light-triggered ATPase activity ofchloroplasts competitively with ATP, whereas ß-naphthylmonophosphate acts non-competitively. None of the four ß-naphthylphosphates were hydrolyzed by these two ATPase activities. Atconcentrations equal to ADP or ATP, ß-naphthyl di-,tri- or tetraphosphate inhibited these three reactions in theorder; ATPase of isolated CF₁> photophosphorylation>light-triggeredATPase of chloroplasts. The results suggest that the effect of the monophosphate isprincipally on the Pi site(s) and that of the di-, tri- or tetraphosphateis on the adenine nucleotide site(s) on the active center ofCF₁. ¹Part of this work was reported at the 1979 Annual Meeting ofthe Japanese Society of Plant Physiologists (Nagoya, April 7,1979) and the 52nd Annual Meeting of the Japanese BiochemicalSociety (Tokyo, October 7, 1979). This work was supported inpart by Grants-in-Aid for Scientific Research from the Ministryof Education, Science and Culture, Japan (311808 and 311909). (Received November 14, 1979; )     

cDNA Sequences of Three Kinds of {beta}-tubulins from Rice

Complete nucleotide sequences of three kinds of rice ß-tubulincDNA clones (pTUB22, R1623 and R2242) were determined. Southernhybridization indicated that these ß-tubulins consistof one gene family. Using RFLP mapping, these three ß-tubulincDNAs were mapped to different chromosomes indicating at leastthree loci for the ß-tubulin gene. The deduced aminoacid sequences of these cDNAs showed a high similarity to otherplant ß-tubulins. The asparagine residue located atthe 100th amino acid from the Nterminus of plant ß-tubulinswas also conserved with these three ß-tubulins. Thisasparagine is thought to be responsible for the sensitivityagainst rhizoxin, the toxin of the pathogen of rice seedlingblight, Rhizopus sp. a soil-borne microorganism. Expressionof the three ß-tubulin genes was analyzed by Northernblotting and all three clones were expressed in root, the possibletarget tissue of rhizoxin. These results suggest that theseclones are candidates of ß-tubulins targeted by rhizoxin.     

Endo-1,4-{beta}-Glucanase in the Cell Wall of Stems of Auxin-Treated Pea Seedlings

Endo-1,4-ß-glucanase induced by treatment of pea seedlingswith 2,4-D was extracted from a preparation of the walls ofepicotyl cells. The ß-glucanase was purified by chromatographyon DEAE-cellulose, affinity chromatography on Con A-Sepharoseand SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The activityof ß-glucanase was retained after removal of SDS andextraction from polyacrylamide gels. The band of a protein (46kDa), that corresponded to the activity of endo-1,4-ß-glucanase,was injected directly into mice for preparation of antiserumand the protein was also subjected to amino acid sequencingafter blotting onto a membrane. Western blot analysis showedthat the antiserum obtained bound to a 46-kDa polypeptide andrecognized endo-1,4-ß-glucanase. The N-terminal sequenceof the 46-kDa polypeptide revealed some homology to abscissionendo-1,4-ß-glucanases of bean and avocado fruit. (Received September 29, 1993; Accepted January 20, 1994)     

Studies on {beta}-Glucosidase Production in Cultured Tissue of Trifolium repens L.

Variation observed in the growth and cytology of white clovercells is considered in relation to enzyme production in culturedtissue. Variation also occurred in the production of ß-glucosidaseby groups of cultured cells derived from the same plant andwith identical cultural histories. This variability betweencultures grown in a completely defined medium, provided an approachto the investigation of the factors affecting ß-glucosidaseproduction in cultured cells. Evidence was obtained from Michaelisconstants that two distinct ß-glucosidases were producedby cultured tissue.     

The Expression of Beta ({beta}) Keratins in the Epidermal Appendages of Reptiles and Birds

The integuments of extant vertebrates display a variety of epidermalappendages whose patterns, morphology and terminal differentiation(epidermal keratins) depend upon interactions between ectodermal(epidermis) and mesodermal (dermis) tissues. In reptiles andbirds, appendage morphogenesis precedes terminal differentiation.Studies have demonstrated that appendage morphogenesis influencesthe expression of the appendage specific keratin genes. However,little is known about the nature of the structural genes expressedby the epidermal appendages of reptiles. How pattern formationand/or appendage morphogenesis influence terminal differentiationof reptilian appendages is not known. The epidermal appendages of reptiles and birds are characterizedby the presence of both alpha () and beta (ß) typekeratin proteins. Studies have focused on the genes of avianß keratins because they are the major structural proteinsof feathers. The occurrence of ß keratin proteinsin the scales and claws of both birds and reptiles and theirimmunological cross-reactivity suggest that the genes for reptilianß keratins may be homologous with those of birds.In bird appendages, the ß keratins are the productsof a large family of homologous genes. Specific members of thisgene family are expressed during the development of each appendage.Recent sequence analyses of feather ß keratins, fromdifferent orders of birds, demonstrate that there is more diversityat the DNA level than was implied by earlier protein sequencingstudies. Immunological techniques show that the same antibodies thatreact with the epidermal ß keratins of the chicken(Gallus domesticus) react with the epidermal ß keratinsof American alligators (Alligator mississippiensis). Furthermore,a peptide sequence (20 amino acids) from an alligator claw ßkeratin is similar to a highly conserved region of avian claw,scale, feather, and feather-like ß keratins. Theseobservations suggest that the ß keratin genes of avianepidermal appendages have homologues in the American alligator.Understanding the origin and evolution of the ß keratingene families in reptiles and birds will undoubtedly add toour understanding of the evolution of skin appendages such asscales and feathers.     

Characterization of serum {beta}-glucuronyltransferase involved in chondroitin sulfate biosynthesis

We studied a glucuronyltransferase involved in chondroitin sulfate(CS) biosynthesis in a preparation obtained from fetal bovineserum by heparin-Sepharose affinity chromatography. This enzymetransferred GlcA from UDP-GlcA to the nonreducing GalNAc residuesof polymeric chondroitin. It required Mn²⁺ for maximal activityand showed a sharp pH optimum between pH 5.5 and 6.0. The apparentKm value of the glucuronyltransferase for UDP-GlcA was 51 µM.The specificity was investigated using structurally definedacceptor substrates, which consisted of chemically synthesizedtri-, penta-, and heptasaccharide-serines and various odd-numberedoligosaccharides with a GalNAc residue at the nonreducing terminus,prepared from chondroitin and CS by chondroitinase ABC digestionfollowed by mercuric acetate treatment. The enzyme utilizeda heptasaccharide-serine GalNAcß1-4GlcAß1-3GalNAcß1-4GlcAß1-3Galß1-3Galß1-4Xylß1-O-Serand a pentasaccharide-serine GalNAcß 4GlcAß1-3Galß1-3Galß1-4Xylß1-O-Seras acceptors. In contrast, neither a trisaccharide-serine Galß1-3Galß1-4Xylß1-O-Sernor an     

Apamin-sensitive, small-conductance, calcium-activated potassium channels mediate cholinergic inhibition of chick auditory hair cells

Acetylcholine released from efferent neurons in the cochlea causes inhibition of mechanosensory hair cells due to the activation of calcium-dependent potassium channels. Hair cells are known to have large-conductance, “BK”-type potassium channels associated with the afferent synapse, but these channels have different properties than those activated by acetylcholine. Whole-cell (tight-seal) and cell-attached patch-clamp recordings were made from short (outer) hair cells isolated from the chicken basilar papilla (cochlea equivalent). The peptides apamin and charybdotoxin were used to distinguish the calcium-activated potassium channels involved in the acetylcholine response from the BK-type channels associated with the afferent synapse. Differential toxin blockade of these potassium currents provides definitive evidence that ACh activates apamin-sensitive, “SK”-type potassium channels, but does not activate carybdotoxin-sensitive BK channels. This conclusion is supported by tentative identification of small-conductance, calcium-sensitive but voltage-insensitive potassium channels in cell-attached patches. The distinction between these channel types is important for understanding the segregation of opposing afferent and efferent synaptic activity in the hair cell, both of which depend on calcium influx. These different calcium-activated potassium channels serve as sensitive indicators for functionally significant calcium influx in the hair cell. Accepted: 12 August 1999     

An Ion-stimulated Adenosine Triphosphatase from Bean Roots

A soluble ATP-ase from bean roots was discovered. The enzyme was assayed by measuring the release of inorganic P from ³²P-labelled ATP. The enzyme is strongly stimulated by hoth sodium and potassium ions, in the alkaline pH range. Its characteristics are compared to that of the animal membrnnal ATP-ase which is presumably involved in the transport of ions in animal tissues.     

Threshold environmental concentrations of cations defining the range of roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> L. in freshwater reservoirs

Threshold levels of sodium, potassium, calcium, and magnesium that define the distribution range of roach in fresh water are 0.015–0.019, 0.012–0.015, 0.006–0.009, and 0.002–0.003 mmol/L, respectively. A reduction in water mineralization is accompanied by a significant decrease in the concentration gradient of cations between the organism of a fish and the environment and by the increased stress on the systems responsible for salty exchange. Many reservoirs are characterized by lower concentrations of potassium in the water than is needed for the success of the roach population. The survival of roach in these waterbodies depends on the amount of potassium consumed by fish with their food. A comparative analysis is performed to assess the threshold levels of cations for roach, two bivalve mollusk species, crayfish, and the filamentous algae Spirogyra.     

Purification and Characterization of Wall-bound Exo-l,3-{beta}-D-Glucanase from Barley (Hordeum vulgare L.) Seedlings

A ß-D-glucanase activity hydrolyzing 1,3:1,4-ß-D-glucanwas released from the cell walls of barley by 3M LiCl treatment.It was purified by sequential cation-exchange, gel-filtrationand hydrophobic chromatography. The molecular mass of the glucanasewas 66 kDa as determined by SDS-polyacrylamide gel electrophoresis.Sequence determination of the first thirty amino acids of theN-terminus revealed a high homology of this enzyme to the Pseudomonasl,4-ß-D-glucosidase (56.5%). The purified ß-D-glucanasehas a pH optimum at 5.0, and hydrolyzes oligosaccharides containingß-D-1,3 or ß-D-1,4 linkage. The glucanaseshowed maximum hydrolytic activity toward laminaritetraose,the rate being about two times that of cellotetraose and aboutfour times that of gentiobiose. Polysaccharides such as lichenan,l,3:l,4-ß-D-glucan (from barley), laminarin and pustulanare also hydrolyzed, but not carboxylmethyl-curdlan, carboxymethyl-cellulose,xyloglucan and maltose. The purified ß-D-glucanaseyielded monomeric glucose from laminarihexaose, and exhibitedcharacteristics of an exo-l,3-ß-D-glucanase (EC 3.2.1.58 [EC] ).The activity and biochemical characteristics of this enzymesuggest that it is an exo-l,3-ß-D-glucanase involvedin the rapid turnover of l,3:l,4-ß-D-glucan in barleycell walls during seedling growth. (Received September 24, 1996; Accepted December 9, 1996)