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1.
Mannose specific lectins of Vicia tetrasperma were purifiedby affinity chromatography with Sephadex G-100, and ion exchangechromatography. Chromatofocusing using PBE-94 gel was successfullyemployed to separate the major isolectins, lectin I and II.Both lectins had the same molecular weight of 78,000 and weretetramers composed of a uniform subunit with a molecular weightof 18,700. Amino acid compositions of these lectins were quitesimilar to each other, rich in aspartic acid (and/or asparagine)and hydroxyl amino acids, and lacking methionine and cysteine.Agar gel double diffusion using anti V. tetrasperma lectin antiserumrevealed that lectins from V. cracca, Pisum sativum, and Lensculinaris, all of which have mannose binding properties, wereantigenically identical. The antiserum reacted with the analogouslectins from V.faba, V. hirsuta, and V. angustifolia, but formationof a spur in the diffusion assay showed that they were slightlydifferent from V. tetrasperma lectin. (Received December 24, 1985; Accepted March 12, 1986)  相似文献   

2.
Three mannose-specific lectins exhibiting considerable similarities in NH2-terminal amino acid sequence were isolated from leaves of the Chinese daffodil Narcissus tazetta (Family Amaryllidaceae). The purification protocol involved extraction with an aqueous buffer, anion exchange chromatography on DEAE-cellulose using stepwise elution with increasing salt concentrations, affinity chromatography on mannose-agarose, and FPLC-gel filtration on Superose 12. From the peak unadsorbed on DEAE-cellulose, and two peaks adsorbed on the ion exchanger and eluted respectively with 0.2 M Tris-HCl buffer and 0.5 M NaCl, were prepared fractions which yielded isolectins 1, 2, and 3 after adsorption on mannose-agarose and FPLC-gel filtraton. All three isolectins were homodimers with a molecular weight of 26 kDa. The lectin unadsorbed on DEAE-cellulose had the lowest, while the most strongly adsorbed lectin had the highest hemagglutinating activity.  相似文献   

3.
Hapten-inhibition studies showed that 3-O-methyl-D-glucose andmethyl--D-mannopyranoside, which are strong sugar haptens ofhemagglutination by pea seed lectins, inhibited (a) bindingof pea seed lectins with Rhizobium leguminosarum J357 cells,(b) the precipitin reaction of pea seed lectins with a capsularpolysaccharide from J357 cells and (c) adsorption of J357 cellsto a pea root. When the capsular polysaccharide was absorb edby the lectins or oxidized by periodate to remove the precipitinreactivity with the lectin, the inhibitory activity of the capsularpolysaccharide towards the adsorption disappeared. I.ectins,which were isolated from the bathing solution of pea roots inacid buffer (pH 2.1), were similar to the seed lectins in sugar-bindingspecificity. The possible existence of lectins on pea root hairsurface was shown by the indirect imraunofluorescent antibodytechnique in combination with the biotin-avidin system. Theseresults suggest that host recognition in Rhizobium-pea. symbiosisis based on the interaction between rhizobial cells and hostlectins. 1Present address: Nodai Research Institute, Tokyo Universityof Agriculture, Setagaya-ku, Tokyo 156, Japan. (Received February 18, 1981; Accepted May 2, 1981)  相似文献   

4.
1-Aminocyclopropane-1-carboxylate (ACC) oxidase (ethylene-formingenzyme) was isolated from wounded mesocarp tissue of Cucurbitamaxima (winter squash) fruit, and its enzymatic properties wereinvestigated. The enzyme required Fe2+ and ascorbate for itsactivity as well as ACC and O2 as substrates. The in vitro enzymeactivity was enhanced by CO2. The apparent Km value for ACCwas 175 µM under atmospheric conditions. The enzyme activitywas inhibited by sulfhydryl inhibitors and divalent cationssuch as Co2+, Cu2+, and Zn2+. ACC oxidase activity was induced at a rapid rate by woundingin parallel with an increase in the rate of ethylene production.The exposure of excised discs of mesocarp to 2,5-norbornadiene(NBD),an inhibitor of ethylene action, strongly suppressed inductionof the enzyme, and the application of ethylene significantlyaccelerated the induction of the activity of ACC oxidase inthe wounded mesocarp tissue. These results suggests that endogenousethylene produced in response to wounding may function in promotingthe induction of ACC oxidase. (Received January 13, 1993; Accepted April 15, 1993)  相似文献   

5.
Leaves from mature Griffonia simplicifolia plants were examined for the presence of leaf lectins possessing sugar binding specificities similar to the four known seed lectins (GS-I, GS-II, GS-III, GS-IV). Three (GS-I, -II, -IV) of the four known G. simplicifolia seed lectins were present in the leaves. Leaf G. simplicifolia lectins I and IV were similar to the respective seed lectins. Leaf GS-II, however, was composed of two types of subunits (Mr = 33,000 and 19,000), whereas the seed lectin consists of only one type of subunit (Mr 32,500). Seed and leaf GS-II lectins also had different isoelectric points. All leaf and seed lectins were similar with respect to their hemagglutination and glycoconjugate precipitation properties and all subunits contained covalently bound carbohydrate. Leaf GS-IV appeared slightly under-glycosylated compared to seed GS-IV.

The fate of GS-I and GS-II seed lectins in aging cotyledons was investigated. GS-I isolectins usually contain isolectin subtypes associated with each main isolectin. Upon inbibition and germination, these GS-I isolectin subtypes disappeared. Over time, GS-II lectin did not change its disc gel electrophoretic properties.

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6.
Tyler  Germund 《Annals of botany》1993,71(4):295-301
Germination, establishment and growth of Melica ciliata L.,an 'acidifuge' species of rocky habitats in Europe, were studiedexperimentally and related to chemical properties of the soilsolution, including pH, base cation composition, Al concentrationand speciation, and Mn concentration. M. ciliata failed to establishin acid leptite soil. However, it was able to grow in solutionat pH 3·6 and exhibited vigorous growth at pH 3·9,a typical soil solution pH of leptite sites, which Melica isunable to colonize. Higher concentrations of Mn than those measuredin any leptite soil solutions did not influence growth. Exposureto 0·037 mmol l-1 (1 mg l-1) of Al3+, a concentrationusually exceeded in the soil solution of leptite sites, severelyretarded root growth. Speciation technique applied to Al insoil solutions obtained by centrifugation demonstrated a closerelationship between H+ and Al concentrations and, in particular,between H+ and free ionic (quickly reacting) Al species. Soilsolution concentrations of free ionic Al proved to be < 0·002mmol l-1 in sites lacking Melica , but often > 0·10mmol l-1 in site lacking Metalica . It is concluded that theinability of M. ciliata to establish in acid soils is not primarilydue to the high H+ concentration but to the high soil solutionconcentrations of Al, especially free Al ionic species at lowsoil pH.Copyright 1993, 1999 Academic Press Melica ciliata, distribution, soil solution, pH, aluminium speciation, manganese, base cations, iron  相似文献   

7.
Peanut lectin was purified from seed meal of the Spanish and Jumbo Virginia varieties of peanut (Arachis hypogaea L.) by affinity chromatography on lactose coupled to Sepharose 4B. Polyacrylamide gel isoelectric focusing resolved the lectin preparation from Jumbo Virginia seeds into seven isolectins (pI 5.7, 5.9, 6.0, 6.2, 6.3, 6.5, and 6.7). Seed meal from the Spanish variety contained six isolectins which were indistinguishable from the pI 5.7, 5.9, 6.2, 6.3, 6.5, and 6.7 isolectins from Jumbo Virginia. Quantitative, lactose-specific hemagglutination was used to examine the lectins in tissues of both peanut varieties. In young (3- to 9-day-old) seedlings of each variety, more than 90% of the total amount of lectins detected in the plants was in the cotyledons. Most of the remainder was in hypocotyls, stems, and leaves; young roots contained no more than 4 micrograms of lectin per plant. Lectins were present in all nonroot tissues of 21- to 30-day-old seedlings, except 27-day-old Spanish hypocotyls. As cotyledons of each variety senesced, several of the more basic isolectins decreased to undetectable levels, but the acidic isolectins remained until at least 15 days after planting. Some of the seed isolectins and several apparently new lactose-binding lectins were also identified in affinity-purified extracts of 5-day-old roots and hypocotyls. Rabbit antibodies raised against the Jumbo Virginia seed isolectin preparation reacted with seed, cotyledon, and hypocotyl lectin preparations from both varieties. Analysis of seed lectin preparations from seven varieties of A. hypogaea and of a related species (A. villosulicarpa) indicated that isolectin composition in Arachis may be a characteristic of both the species and the subspecies (botanical type) to which the variety belongs.  相似文献   

8.
Ion currents across the plasma membrane of the unicellular greenalga Eremosphaera viridis were characterized with electrophysiologicalmethods, especially the two electrode voltage-clamp-technique.Under different conditions, at increased external Clconcentrations or after perfusion of different anion channelblockers (A9C (anthracen-9-carboxylic acid), NPPB ((5-nitro-2-3-phenylpropylamino)benzoic acid) and ZnCl2), increased instantaneous negative currentswere observed. The negative currents were carried by cationfluxes into the cell. The transporter responsible had low selectivityamong potassium and sodium. Additionally also divalent cationswere transported. The cation influx was not affected by thepotassium channel inhibitors TEA (tetraethylammoniumsulfate),Ba2+ or Cs+ at concentrations of 1 mM, but was strongly reducedby 100 µM AlCl3. Our results with E. viridis demonstrate,for the first time for an unicellular alga, the existence ofan inwardly rectifying cation current across the plasma membrane.Parallels and differences to inwardly rectifying cation currentsand channels described in plasma membranes of other plant cellsare discussed. (Received May 10, 1993; Accepted September 13, 1993)  相似文献   

9.
The internal salt content and distribution in photosynthetictissues as well as the effect of NaCl on photosynthetic carbonfixation enzymes was investigated in two seagrass species fromthe Red Sea. Concentrations of both Na+ and Cl were lower in the chloroplast-richepidermis than in underlying cell layers in Halophila stipulacea.In Halodule uninervis, the concentration of Na+ was lower inthe epidermis than in the underlying cells, while K+ was evenlydistributed between cell layers. The epidermal concentrationsof Na+ were estimated to be 0.17 and 0.10 M for Halophila stipulaceaand Halodule uninervis, respectively, which were about to the average leaf concentrations. Epidermal Cl concentrationof Halophila stipulacea was estimated to be 0.08 M, a valueonly about of the overall leaf concentration. Phosphoenolpyruvate carboxylase (PEPcase) extracted from leavesof these seagrasses showed increased activity at 0.05–0.3M NaCl in vitro. Ribulose-l, 5-bisphosphate carboxylase (RuBPcase)activity, on the other hand, was inhibited by NaCl at all testedconcentrations. At epidermal NaCl concentrations, PEPcase activitywas thus stimulated while RuBPcase was inhibited. The reducedRuBPcase activity at such concentrations compared to salt-freeconditions was still sufficient to account for observed photosyntheticrates. We conclude that these seagrasses have adapted to a saline environmentboth by maintaining relatively low ion concentrations in theepidermis where photosynthesis occurs and by having carbon-fixingenzymes capable of functioning in the presence of salt.  相似文献   

10.
Maize (Zea mays L., hybrid Cargill 147) seedlings, grown inaseptic conditions, were inoculated with three strains of Azospirillumlipoferum (Al op 33, Al iaa 320, and ATCC 29708) or culturedin different concentrations of indol-3-acetic acid (IAA) orgibberellin A3 (GA3). After 48 h, root length, root surfacearea, root dry weight, and shoot dry weight were measured inall treatments. Gibberellin content was evaluated in selectedroots of control and Azospirillum inoculated seedlings by gaschromatography-mass spectrometry-selected ion monitoring withthe use of deuterated gibberellins as internal standards. Thethree strains of A. lipoferum, IAA (2 ng ml–1), and GA3(40 to 400 pg ml–1) significantly enhanced root growth.Improvement of root hair growth and density was obtained mainlywith A. lipoferum strain Al op 33 and GA3 40 pg ml–1.GA3 was identified by gas chromatography-mass spectrometry (byboth, full scan and selected ion monitoring) in a free acidfraction from roots of the seedlings inoculated with A. lipoferum.In the roots of the non inoculated seedlings GA3 was found afterhydrolysis of a fraction expected to contain glucosyl conjugates. (Received April 26, 1993; Accepted September 27, 1993)  相似文献   

11.
Treatment of mitochondria isolated from Texas male sterile cytoplasmcorn (T mitochondria) with high concentrations of dicyclohexylcarbodiimide(DCCD) (140 nmol DCCD mg–1 mitochondrial protein) completelyand immediately inhibited T mitochondrial swelling by Helminthosporiummaydis Race T toxin (HmT toxin). In order to obtain a specificinteraction between DCCD and the ATPase complex T mitochondriawere incubated with lower DCCD concentrations (1–5 nmolDCCD mg–1 mitochondrial protein) for up to 8 h at 4 °C.After 8 h incubation in the presence of 3.75 nmol DCCD mg–1mitochondrial protein, toxin-induced swelling was decreasedby 69%. Specificity of DCCD action upon the ATPase complex wasconfirmed by (1) SDS gel electrophoresis and fluorographic analysesof proteins from [14C]-DCCD-treated T mitochondria and immunoprecipitatesand (2) physiological experiments showing that DCCD exertednone of its other documented effects. These data suggest thatHmT toxin interacts with the ATPase complex of T mitochondriaeither at or near the DCCD-binding protein within the membranesector of the complex. Key words: Zea mays L., Helminthosporium maydis, Mitochondria  相似文献   

12.
The viscous and elastic moduli at different shear rates, togetherwith various biological oceanographic properties, were determinedin seawater from different hydrological layers in the southernNorth Sea in June. The biological oceanographic parameters includedPhaeocystis and Noctiluca abundances, chlorophyll a level (Chl),bacteria. HNAN and aggregate volume fraction. The plankton wasjointly dominated by Phaeocyslis sp. and Noctiluca scinullans.Noctiluca abundance showed no correlation with any other biologicalor viscoelastic parameter, but Phaeocystis abundance correlatedstrongly. The other biological parameters correlated with Phaeocystisand with each other positively and mostly significantly. Overall,viscoelasticity correlated more strongly with Chl than withany other biological parameter. For non-microlayer samples,the excess complex (viscoelastic) modulus (µ.Pa) G*E =2.0 x Ch11–3 (Chl in mg m–3). Viscous and elasticmoduli also correlated closely with each other. For a givenvalue of Chl. the microlayer samples were 6.5 or 14 times (dependingon the estimation method) more viscoelastic than in bulk-phasesamples. Viscoelasticity in samples of settled benthic ‘fluff’were lower even than bulk-phase samples, but this differencewas not significant. Comparison with Mediterranean data on viscoelasticity(Jenkinson. Oceanol. Acta, 16, 317–334, 1993), using publishedvalues for phytoplankton biomass (Wiadnyana, J. Rech. Océanogr.,17, 1–6, 1992), suggests that the relationship betweenChl (or phytoplankton biomass) and viscoelasticity might begeneral. This apparent biomodification of the viscosity andelasticity of seawater is discussed in relation to its likelyimpact on turbulence and plankton ecology.  相似文献   

13.
《FEBS letters》1985,184(1):104-109
The complete amino acid sequences of the β1- and β2-subunits of the isolectins (LoL1 and LoL11) from seeds of Lathyrus ochrus were determined by analysis of peptides derived from the proteins by digestion with trypsin, chymotrypsin, pepsin and the S. aureus V8 protease, as well as fragments produced by cleavage with iodosobenzoic acid. Both β-subunits consisted of singlepolypeptide chains of 181 amino acids, which differed from one another in only 3 positions. The homology of the Lathyrus ochrus isolectins with the other two-chain lectins of the tribe Vicieae, and the single-chain lectins of other tribes of the Leguminosae is discussed.  相似文献   

14.
In shake-flask culture asymmetrin was produced by PenicIlliumthomiiaud Byssochlamys niveaduring autolysis. A bioassay wasdevised to estimate relative concentrations of this compound.Culture filtrates of P. thomiicontaining asymmetrin caused adecrease in wet weight, dry weight, and total nitrogen of Phaseolusvulgarisup to 18 days after treatment. At certain intervalsafter treatment with culture filtrate of B. nivea,respirationwas inhibited, and changes were observed in isotope ratio valuesof plants supplied with glucose-14 C. Plants treated previouslywith culture filtrates of B. nivearesponded but little to gibberelhcacid. High concentrations of mdoleacetic acid inhibited growthof control plants but stimulated growth of plants treated withculture filtrate of B. nivea. 1Present address: United Fruit Company, Norwood, Massachusetts.Supported in part by a Public Health Service Fellowship No.GF 13,776 from the division of General Medical Sciences, PublicHealth Service, National Institutes of Health, and Grant G 20989from the National Science Foundation. 2Journal Paper No. 2170 of the Purdue Agricultural ExperimentStation.  相似文献   

15.
We previously reported that the red alga Eucheuma serra contains large amounts of mitogenic isolectins (ESA-1 and ESA-2), the hemagglutinating activities of which were strongly inhibited by glycoproteins bearing high mannose-type N-glycans. We therefore further examined two other species, E. amakusaensis and E. cottonii. Several lectins were isolated easily by a combination of extraction with aqueous ethanol, precipitation with cold ethanol, gel filtration, and ion exchange chromatography from both species, respectively. The purified lectins were designated as EAA-1, EAA-2, EAA-3, ECA-1 and ECA-2 after the specific names of both algae. The yields of EAAs and ECAs were as high as 2.8 and 2.7 mg g−1 of dry tissue, respectively, indicating that both species would also be good sources for high lectin yields. The five purified lectins shared the same properties in hemagglutinating activity, mitogenic activity, and hemagglutination-inhibition test in which glycoproteins bearing high mannose-type N-glycans were the most inhibitory. They also had almost identical molecular weight and 20 N-terminal amino acid sequence to each other and to those of ESAs, and only differed in the isoelectric point, indicating that they are isolectins to each other. The study thus demonstrated that several species of Eucheuma contain high yields of lectins homologous between species, suggesting that the genus as a whole may be considered as a valuable source of lectin proteins. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

16.
Plasma Membrane H+-ATPase in Guard-Cell Protoplasts from Vicia faba L.   总被引:2,自引:0,他引:2  
The activity of plasma membrane H+-ATPase was measured withmembrane fragments of guard-cell protoplasts isolated from Viciafaba L. ATP hydrolytic activity was slightly inhibited by oligomycinand ammonium molybdate, and markedly inhibited by NO3and vanadate. In the presence of oligomycin, ammonium molybdateand NO3, the ATP-hydrolyzing activity was strongly inhibitedby vanadate. It was also inhibited by diethylstilbestrol (DES),p-chloromercuribenzoic acid (PCMB) and Ca2+, but slightly stimulatedby carbonyl cyanide m-chlorophenylhydrazone (CCCP). The acitivityhad higher specificity for ATP as a substrate than other phosphoricesters such as ADP, AMP, GTP and p-nitrophenylphosphate; theKm was 0.5 mM for ATP. The activity required Mg2+ but was notaffected by K+, and it was maximal around pH 6.8. When guard-cellprotoplasts were used instead of membrane fragments, the ATPaseactivity reached up to 800µmol Pi.(mg Chl)–1.h–1in the presence of lysolecithin. These results indicate thatthe guard cell has a high plasma membrane H+-ATPase activity. (Received December 23, 1986; Accepted April 28, 1987)  相似文献   

17.
Cellular membrane fractions, including endoplasmic reticum (ER),Golgi-enriched membrane, plasma membrane and tonoplasts, wereisolated from Vigna radiata seedlings. Each of these membranefractions was associated with specific ATPases which were highlydependent on Mg2+. ATPases of ER, Golgi-enriched membrane andplasma membrane were sensitive to vanadate but the tonoplastATPase was not. ATPases were mostly dependent on Cl1, but aslight stimulation by K+ was observed in the case of ATPasesof Golgi-enriched membrane and plasma membrane. KNO3 inhibitedtonoplast ATPase but stimulated the other ATPases. ER ATPasecan be distinguished from other ATPases by the following characteristics:specific inhibition by KNO2 and Triton X-100, stimulation bylow concentrations of diethylstilbestrol and 4,4'-diisothiocyanostilbene-2,2'-disulfonicacid, and high sensitivity to heat. The ATPases showed typicalMichaelis-Menten kinetics and had Km values of 0.5 to 0.6 ITIMMg2+-ATP for ER, Golgienriched-membrane and tonoplast ATPases,and 2.27 msi Mg2+-ATP for plasma membrane ATPase. ATPases ofGolgi-enriched membranes and plasma membranes had similar properties,but they were still distinguishable by the differences in theirKm values and their responses to Triton X-100. Based on theseresults, it is postulated that each cellular membrane is associatedwith a specific ATPase in cells of V. radiata. 1Contribution No. 3171 from the Institute of Low TemperatureScience. (Received April 22, 1988; Accepted September 28, 1988)  相似文献   

18.
Lectin-induced apoptosis of tumour cells   总被引:3,自引:0,他引:3  
The mechanisms of cytotoxic activity of Griffonia simplicifolia1-B4 (GS1B4) and wheat germ agglutinin (WGA) lectins againstvarious murine tumour cell lines were studied. Tumour cellsthat lack lectin-binding carbohydrates were resistant to lysisby these lectins. However, YAC-1 cells that expressed GS1B4lectin-binding sites showed low sensitivity to lysis. To furtheranalyse the relative importance of cell surface carbohydratesin lectin cytotoxicity, BL6–8 melanoma cells, which donot express the  相似文献   

19.
Neill, S. J., McGaw, B. A. and Horgan, R. 1986. Ethylene and1-aminocyclopropane-l-carboxylic acid production in flacca,a wilty mutant of tomato, subjected to water deficiency andpretreatment with abscisic acid —J. exp. Bot. 37: 535–541. Plants of Lycoperstcon esculentum Mill. cv. Ailsa Craig wildtype and flacca (flc) were sprayed daily with H2O or 2?10–2mol m–3 abscisic acid (ABA). ABA treatment effected apartial phenotypic reversion of flc shoots; leaf areas wereincreased and transpiration rates decreased. Leaf expansionof wild type shoots was inhibited by ABA. Indoleacetic acid (IAA), ABA and l-aminocyclopropane-l-carboxylicacid (ACC) concentrations were determined by combined gas chromatography-massspectrometry using deuterium-labelled internal standards ABAtreatment for 30 d resulted in greatly elevated internal ABAlevels, increasing from 1?0 to 4?3 and from 0?45 to 4?9 nmolg–1 fr. wt. in wild type and flc leaves respectively.Endogenous IAA and ACC concentrations were much lower than thoseof ABA. IAA content ranged from 0?05 to 0?1 nmol g–1 andACC content from 0?07 to 0?24 nmol g–1 Ethylene emanationrates were similar for wild type and flc shoots. Wilting of detached leaves induced a substantial increase inethylene and ACC accumulation in all plants, regardless of treatmentor type. Ethylene and ACC levels were no greater in flc leavescompared to the wild type. ABA pretreatment did not preventthe wilting-induced increase in ACC and ethylene synthesis. Key words: ABA, ACC, ethylene, wilting, wilty mutants  相似文献   

20.
Eight cvs of barley (Hordeum vulgare L.) were separately plantedwith Wild Oats (Avena fatua L., genetically pure line CS40)in a sand culture with two external K+ concentrations. Substantialdifferences were observed among barley cvs in their abilityto compete with wild oat. The variety Fergus was highly competitiveat both high and low [K+]e, whereas Steptoe was competitiveonly at high [K+]e, and Compana was only weakly competitivewith wild oat. The differences between barley cvs were relatedto their previously reported efficiencies of K+ uptake and utilization. Hordeum vulgare L., Avena fatua L., barley, wild oat, competition, K+ nutrition, utilization efficiency  相似文献   

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