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The localization of phenylalanine ammonia-lyase [EC 4.3.1.5] within sweet clover (Melilotus alba) leaves was investigated. Apical buds and axillary leaves contained 15 to 30 times more enzyme activity than did mature leaves. Mesophyll protoplasts were prepared by digesting young leaves with Cellulysin and Macerase and ...
The properties of homoserine dehydrogenase (EC 1.1.1.3) isolated from shoots of young etiolated seedlings of Zea mays L. var. earliking can be reversibly altered by dialysis against an appropriate buffer. Treatment with 500 millimolar potassium phosphate buffer (pH 7.5) in the absence of l-threonine results in diminish...
Upon illumination of dark-grown maize seedlings (5 days old) with incandescent light, there occurred a nearly simultaneous increase, after a certain lag period, in the activities of enzymes engaged in the C(4) pathway and the Calvin-Benson cycle. The light-induced biosynthesis of chlorophyll (a and b) precedes the incr...
Two endo-1,3,-beta-d-glucanases (I and II, EC 3.2.1.6) are present in etiolated peas at opposite ends of the stem. Glucanase I from subapical regions degrades substrates to a series of low molecular weight dextrins, and is most readily assayed reductometrically (e.g. as laminarinase). Glucanase II from basal regions pr...
SUCCINATE DEHYDROGENASE (SUCCINATE: phenazine methosulfate oxidoreductase, EC 1.3.99.1) activity in crude mitochondrial fraction from pea (var. Alaska) cotyledons increased during seed imbibition to reach a maximum after about 12 hours. The increase was not inhibited by either cycloheximide or d(-)threo-chloramphenicol...
The activities of three enzymes of phenolic biosynthesis and six of general metabolism were studied at 24-hour intervals between the 3rd and 8th day after planting in barley shoots treated with the chlorosis-inducing herbicide Sandoz 6706 and grown in the dark or under high or low intensity light. The herbicide had no ...
Scutella separated from grains of Himalaya barley after germination for 3 days rapidly took up l-leucine from aerated incubation media; with 1 millimolar leucine the rate varied between 4 and 14 micromoles per gram per hour and the pH optimum was at 3.5 to 5, both depending on buffer composition and prewashing time. Th...
Boron is required for fiber growth and development in cotton ovules cultured in vitro. Incorporation of [(14)C]glucose by such fiber from supplied UDP-[(14)C]glucose into the hot alkali-insoluble fraction is rapid and linear for about 30 minutes. Incorporation of [(14)C]glucose from such substrate by fibers grown in bo...
Submitochondrial particles freshly prepared by sonication from pea cotyledon mitochondria showed low ATPase activity. Activity increased 20-fold on exposure to trypsin. The pea cotyledon submitochondrial particle ATPase was also activated by "aging" in vitro. At pH 7.0 addition of 1 millimolar ATP prevented the activat...
Subcellular organelles from castor bean (Ricinus communis) endosperm were isolated on discontinuous sucrose gradients from germinating seeds which were 1 to 7 days postimbibition. Marker enzyme activities of the organelles were measured (fumarase, catalase, and triose phosphate isomerase) and the homogeneity of the org...
The CO(2) compensation point of the submersed aquatic macrophyte Hydrilla verticillata varied from high (above 50 microliters per liter) to low (10 to 25 microliters per liter) values, depending on the growth conditions. Plants from the lake in winter or after incubation in an 11 C/9-hour photoperiod had high values, w...
During grain formation stage (90 to 110 days), the youngest flag leaf of rice (Oryza sativa L. cv. Jaya) remained metabolically most active (as indicated by cellular constituents and enzyme activities) and the third leaf the least active. At the grain development stage (110 to 120 days) the above pattern of age-related...
Freshly isolated protoplasts from leaves of oat seedlings (var. Victory) which do not divide when cultured on a wide range of media are capable of incorporating tritiated leucine, uridine, and thymidine into trichloroacetic acid-insoluble macromolecules. Over 70% of the leucine and uridine incorporated over an 18-hour ...
Pear fruits (Pyrus communis L. var. Bartlett) were treated with solutions containing aminoethoxyvinylglycine (AVG) using a modified vacuum infiltration method that introduced 4.3 milliliters solution per 100 grams tissue. At concentrations of 1 millimolar, AVG strongly inhibited ethylene production and delayed for 5 da...
Xylem extracts of poplar tree contained glyoxylate reductase specific for NADPH. By isoelectric focusing in the pH ranges 3.5 to 10 or 4 to 6, the enzyme exhibited a single peak of activity at pH 5.4. The enzyme showed essentially no activity toward hydroxypyruvate, pyruvate, or NADH. The reaction was optimal at pH 6.0...
Nitrogen deficiency and the presence of specific organic carbon sources prevent chloroplast development in Euglena. In exponentially growing cultures, chlorophyll levels were low and independent of the nitrogen content of the growth medium. Chlorophyll levels increased in stationary phase and the amount of chlorophyll ...
Four chlorophyll-protein complexes have been resolved from the cyanophyte, Nostoc sp., by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis at 4 C. Complexes solubilized by SDS from Spinacia oleracea were run for comparison. As has been well documented, the P700-chlorophyll a-protein complex from the high...
The activities of catalase, peroxidase, indoleacetic acid (IAA) oxidase and peroxide levels in cucumber plants during and after chilling were determined. During 96 hours at 5 C and 85% relative humidity, catalase activity declined, IAA oxidase activity increased, and peroxide concentrations increased. Peroxidase activi...
An acetylcholine hydrolyzing enzyme was prepared and purified (40 times) from dwarf bean hypocotyl hooks. The purity of the enzyme was proved by polyacrylamide gel electrophoresis. The molecular weight of the enzyme was determined to be 65,000 daltons. Enzyme activity was the highest at pH 8.0 and between 30 and 36 C. ...
An assay was developed for simultaneous kinetic analysis of the activities of the bifunctional plant enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase [EC 4.1.1.39]. [1-(14)C,5-(3)H]Ribulose 1,5-bisphosphate (RuBP) was used as the labeled substrate. Tritium enrichment of the doubly labeled 3-phosphoglycerate (3-PG...
Evidence in support of the hypothesis of gene expression and subunit association suggested earlier for Triticum alcohol dehydrogenase has been obtained through purification and partial characterization of the enzyme from tetraploid wheat. Three isozymes of alcohol dehydrogenase were separated and purified to apparent h...
Incubation of excised, 1-centimeter long root tips of corn in chilled distilled H(2)O increases the activity of extractable GDP fucose: polysaccharide fucosyl transferase. Enhancement of transferase activity is optimal at 10 C and incubation at this temperature for 8 hours causes a 3- to 5-fold stimulation of activity....
Homogenates of mung bean cotyledons were subjected to equilibrium density centrifugation on linear sucrose gradients and the positions of the various organelles determined by assay of marker enzymes. Measurement of phospholipid distribution on such gradients showed that the major peak of phospholipid at a density of 1....
Oxyleghemoglobin was used to supply low concentrations of O(2) to H(2)-oxidizing bacteroids from Rhizobium japonicum USDA 122 DES. The H(2) oxidation system of these bacteroids was capable of effectively utilizing O(2) at the low concentrations of O(2) expected to be found in soybean nodules. Apparent K(m) values of ap...
Four legume species (four genera) were examined and found to contain hemagglutinins with properties similar to those which we have previously described for the enzymic hemagglutinin in Vigna radiata. Examination of extracts by gel filtration and ion exchange chromatography showed that an alpha-galactosidase activity ex...
Differences in the accumulation of total seed protein and globulin-1 (G1) protein were detected among three inbred lines of common bean. Total protein accumulation ranged from 2.3 to 3.7 milligrams per cotyledon pair per day among lines. In all lines the dry weight and protein accumulation ceased and a loss of chloroph...
Placing light-grown Lemna gibba L. G-3 into the dark results in a changed pattern of protein synthesis. Although the amount of protein in the tissue and the over-all rate of incorporation of [(35)S]methionine into protein does not significantly decline during four days of darkness, the rate of synthesis of three polype...
Pea cotyledons were injected with d-[(14)C]mannose or d-[(14)C]-glucosamine and incubated for 1 to 1.5 hours. Cotyledons were homogenized and subcellular fractions were isolated by differential centrifugation followed by linear sucrose density gradient centrifugation.Radioactivity that was precipitated by trichloroacet...
Two proteases active in and specific to oat etioplasts and up to 24-hour etiochloroplasts, only very slightly contaminated by other cellular compartments are described. The enzyme showed pH optima of 4.2 (acid) and 6.8 (neutral), temperature optima of 50 C and the highest level of enzyme activity was with prolamellar b...
Electron donation to photosystem I was studied in highly resolved particles from spinach. Divalent cations increased the efficiency of electron donation from spinach plastocyanin to P700(+) through a decrease in the apparent K(m) for plastocyanin. Cytochrome f was not an efficient electron donor for P700(+) in the pres...
A protein kinase activity from spinach chloroplasts, tightly associated with the thylakoid membranes, has been solubilized and partially characterized. This membrane-bound protein kinase is stimulated by light and electron transport activity through photosystem II appears to be required for stimulation.Electron transpo...
Brief saturating light flashes were used to probe the mechanism of inactivation of O(2) evolution by Tris in chloroplasts. Maximum inactivation with a single flash and an oscillation with period of four on subsequent flashes was observed. Analyses of the oscillations suggested that only the charge-collecting O(2)-evolv...
Photosynthesis and the Warburg effect (O(2) inhibition of photosynthesis) were evaluated in preparations of intact spinach chloroplasts enriched with varying amounts of lysed chloroplasts. Increasing the ratio of broken to intact plastids resulted in decreased rates of CO(2) assimilation.Hydrogen peroxide when added at...
The localization of some key enzymes leading to sucrose synthesis in photosynthetic tissue of C(3) and C(4) species was investigated. These included UDP-glucose (UDPG) pyrophosphorylase, sucrose phosphate synthetase, and glycerate kinase. Whether glycerate kinase is localized exclusively in the chloroplast or partly ou...
Enzymes dissociated from corn (hybrid B73 x Mo17) seedling cell walls by solutions of high ionic strength possess the capacity to degrade Avena caryopsis glucan. Inhibitor studies disclosed that both endo- and exoenzyme activities were involved and that the reaction sequence paralleled the autolytic solubilization of b...
Light and dark (14)CO(2) assimilation, pulse-chase ((14)CO(2) followed by (12)CO(2)) labeling experiments both in the light and in the dark, photorespiratory activity and some enzymes (ribulose 1,5-bisphosphate (RuBP) carboxylase, phosphoenolpyruvate (PEP) carboxylase, and NADP-malic enzyme) were followed in sections o...
Ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) was crystallized from a heterozygous tobacco (Nicotiana tabacum L.) aurea mutant (Su/su), its wild-type sibling (su/su), and green revertant plants regenerated from green spots found on leaves of haploid Su plants. No differences were found in the specific a...
The poly(A)-containing mRNA from barley aleurone layers pretreated with gibberellic acid has been purified by phenol-chloroform extraction and repeated oligo[d(pT)]-cellulose chromatography. This RNA has been translated in both the wheat germ and reticulocyte lysate in vitro translation systems with greater than 50% of...
Assays of alkaline pyrophosphatase activity in crude plant extracts are inhibited by soluble calcium coextracted with the enzyme from leaf tissues. Calcium concentrations in most extracts are high enough to interfere seriously with the assay.All C(4) plants examined kept their soluble calcium and interference with pyro...
Nicotinamide adenine dinucleotide phosphate (NADP)-dependent glyceraldehyde-3-phosphate dehydrogenase (GPDH) (EC 1.2.1.13), a chloroplast enzyme, had low activity in etioplasts of maize leaves. A light dependent increase of enzyme activity of 7-day-old etiolated seedlings showed a lag period of about 2.5 hours followed...
Photoautotrophic and mixotrophic growth of Lemna paucicostata Hegelm. 6746 (formerly Lemna perpusilla Torr. 6746) was investigated to establish standardized conditions for biochemical studies. Optimal temperature for growth was 29 to 30 C. The medium used previously (Datko AH, Mudd SH, Giovanelli J 1977 J Biol Chem 252...
The sequence of frond emergence and the intervals required for daughter colony separation have been determined for Lemna paucicostata Hegelm. 6746 growing under standardized conditions. After separation of a new mother colony, the first daughter colony is produced from the left meristematic pocket and separates after a...
Synthesis of nitrate reductase (EC 1.6.6.1) in Chlorella vulgaris was studied under inducing conditions, i.e. with cells grown on ammonia and then transferred to nitrate medium. Cycloheximide (but not chloramphenicol) completely inhibited synthesis of the enzyme, but only if it was added at the start (i.e. at the time ...
The K(+)-stimulated ATPase was partially purified from a plasma membrane fraction from corn roots (WF9 x Mo 17) by solubilization with 30 millimolar octyl-beta-d-glucopyranoside followed by precipitation with dilute ammonium sulfate. The specific activity of the enzyme was increased about five times by this procedure. ...
A single enzyme catalyzes the synthesis of all four N(2)-(1-carboxyethyl)-amino acid derivatives found in a crown gall tumor tissue induced by Agrobacterium tumefaciens (E. F. Sm. and Town.) Conn strain B6 on sunflower (Helianthus annuus L.). This enzyme, octopine synthase, has been purified by ammonium sulfate fractio...
Treatment of etiolated pea (Pisum sativum (L. cv. Alaska) seedlings with 2'-isopropyl-4'-(trimethylammonium chloride)-5'-methylphenyl piperidine-1-carboxylate (Amo-1618) prior to irradiation with white light inhibits photomorphogenesis and formation and stacking of thylakoid membranes in the chloroplasts, as well as (-...
The enzymic hydrolysis of acyl-coenzyme A occurs in intact and purified chloroplasts. The different components of spinach chloroplasts were separated after a slight osmotic shock and the purified envelope membranes were shown to be the site of very active acyl-CoA thioesterase activity (EC 3.1.2.2.). The enzyme, which ...
Reduced (GSH), oxidized (GSSG), and protein-bound (PSSG) glutathione were determined in dry and hydrated wheat embryos. Dry embryos contained about 0.6 mumoles per gram dry weight each of GSSG and PSSG, and these levels declined 5- to 10-fold within minutes after the onset of imbibition. GSH declined from about 8 to 2 ...
Phospholipase A(2) (Naja naja), the nonpenetrating dye trinitrobenzene sulfonate, and the penetrating dye dinitrofluorobenzene, were used to determine the transmembrane distributions of phospholipids of mitochondria and glyoxysomes isolated from endosperm tissue of castor bean (Ricinus communis L. var. Hale). These stu...
The presence of lipid- and protein-bound sugars in the major organelle fractions isolated from germinating castor bean (Ricinus communis L.) endosperm has been established. Microsomes, glyoxysomes and mitochondria were subfractionated into a membrane fraction and a fraction containing peripheral membrane and soluble ma...
Density-labeling with 10 millimolar K(15)NO(3)/70% (2)H(2)O has been used to investigate isocitrate lyase synthesis during greening of sunflower (Helianthus annuus L.) cotyledons when the glyoxysomal enzyme activities sharply decline and the transition in cotyledonary microbody function occurs. A density shift of 0.005...
The white leaf tissue of seedlings of Zea mays L. affected by the recessive nuclear gene iojap shows no photosynthetic activity; it contains about 1.4% of carotenoid and less than 0.1% of chlorophyll a content of normal green tissue. Neither fraction I protein nor chloroplast adenosine triphosphatase (EC 3.6.1.4) (CF(1...
The typical procedure for determining NAD(+)-malic enzyme (EC 1.1.1.39) is to calculate the enzyme rate to be DeltaA(340)/Delta time after the endogenous NAD(+)-malic dehydrogenase (EC 1.1.1.37) catalyzed reaction has reached equilibrium. This ignores the equilibrium shift of oxaloacetic acid and NADH during the course...
The metabolism of (14)C-labeled aspartic acid, diaminopimelic acid, malic acid and threonine by isolated pea (Pisum sativum L.) chloroplasts was examined. Light enhanced the incorporation of [(14)C] aspartic acid into soluble homoserine, isoleucine, lysine, methionine and threonine and protein-bound aspartic acid plus ...
The membrane components of the castor bean spherosomes were characterized. The storage triacylglycerols of isolated spherosomes were extracted with diethyl ether, and the membrane was isolated by sucrose gradient centrifugation. It had an apparent equilibrium density of 1.12 grams per cubic centimeter, and possessed an...
The kinetics of [(3)H]thymidine incorporation into the DNA of carrot suspension cultures were investigated. At a thymidine concentration of 0.1 micromolar, incorporation into DNA is not quantitative but ceases after only 14% of the thymidine has been incorporated. Thymidine incorporation into DNA is resumed following a...
Allantoin and allantoic acid are the major forms of nitrogen transported from soybean nodules to other parts of the plant. Neither the pathway or the site of ureide synthesis has been demonstrated in root nodules.Bacteroid and cytosol (plant portion) fractions were prepared and the purity of each fraction was determine...
The nuclease activity of developing corn endosperms was found to consist mainly of plant RNase I during the period of major deposition of dry weight. The RNase concentrations in most inbred lines and hybrids increased throughout development, but there were large differences among genotypes in the enzyme levels at all s...
The soluble bean leaf auxin-binding protein (ABP) has a high affinity for a range of auxins including indole-3-acetic acid (IAA), alpha-napthaleneacetic acid, phenylacetic acid, 2,4,5-trichlorophenoxyacetic acid, and structurally related auxins. A large number of nonauxin compounds that are nevertheless structurally re...
Reliable estimates of Michaelis constants (K(m)) and inhibitor constants may be obtained, in the absence of control over the amount of enzyme being added to any assay system, provided the following constraints are met. Michaelis-Menten kinetics are obeyed. Two rate measurements must be made with the same sample of enzy...
The ultrastructure and enzyme distribution in chloroplasts and other subcellular fractions isolated from the siphonous green alga, Caulerpa simpliciuscula, are described. The isolated chloroplasts were similar in appearance to those in the tissue from which they were derived, and in typical preparations 70% or more wer...
Some dynamic aspects of leaf elongation in rice were studied. Under both well watered and water-deficient conditions, leaf elongation rates were 15 to 30% greater during the day than during the night. Night temperatures below 27 C limited the rate of elongation at night but when night temperatures exceeded 27 C, night ...
An extract from cucumber cotyledons was shown to cause an inhibition of protochlorophyll biosynthesis and accumulation. The extract inhibited the net synthesis of protochlorophyll as well as the incorporation of delta-amino[(14)C]levulinic acid into protochlorophyllide and protochlorophyllide ester by excised cotyledon...
Four maize hybrids, two with high and two with low levels of postanthesis nitrate reductase activity were grown under field conditions. The characteristic enzyme patterns had been established in previous work. Nitrate reductase and proteases were measured in three representative leaves (ear leaf, fourth leaf above and ...
The microbodies extracted from the cotyledons of mustard seedlings (Sinapis alba L.) form two bands (at 1.18 kilograms per liter together with the mitochondria, and at 1.24 kilograms per liter) on conventional isopycnic sedimentation density gradients. The artifactual co-banding of part of the microbodies with the mito...
A method for determining the subcellular metabolite levels in spinach protoplasts is described. The protoplasts are disrupted by centrifugation through a nylon net, releasing intact chloroplasts which pass through a layer of silicone oil into perchloric acid while the remaining cytoplasmic components remain over the oi...
Using isolated spinach (hybrid 424) chloroplasts deprived of their envelopes (reconstituted chloroplast system), the metabolism of glucose, glucose 1-phosphate, and glucose 6-phosphate via the oxidative pentose phosphate cycle was analyzed. The activity of oxidative pentose phosphate cycle was monitored by continuous s...
Membrane barriers which prevent direct contact between Fusarium solani and pea endocarp tissue prevent fungal spores from inducing phytoalexin production. Conversely, preinduced host resistance responses are not readily transported from the plant across the membrane barrier to Fusarium macroconidia.Crude enzyme extract...
When leaf blades of fully expanded second leaves of barley (cv. Prior) were excised and incubated with the cut end in a 20 milligram per liter solution of abscisic acid, they accumulated proline at the rate of about 1 micromole per hour per gram fresh weight after a 3- to 4-hour lag. This accumulation occurred reproduc...
The effect of bicarbonate ions on induction of Mg(2+)-ATPase activity, on the N-ethylmaleimide inhibition of phosphorylation and on energy-dependent adenine nucleotide exchange has been examined with pea seedling chloroplasts. Incubation of chloroplasts with N-ethylmaleimide in the presence of 15 millimolar bicarbonate...
ATP, GTP, CoA, Mg(2+), and Mn(2+) did not inhibit biosynthesis of steryl glycoside and acylated steryl glycoside when added singly to enzyme preparations from spinach leaves. The combination of ATP (but not GTP), CoA, and Mg(2+) or Mn(2+) caused marked inhibition, especially of steryl glycoside biosynthesis, when react...
Mung bean plants (Wilczek) accumulate increasingly greater amounts of buffer-extractable copper in both their shoots and roots when grown in liquid medium containing greater than 2 micrograms per milliliter copper (31.4 micromolar) as cupric sulfate. This increase in soluble copper is accompanied by an increase in the ...
Untreated mung bean hypocotyls produced very little C(2)H(4) but, upon treatment with 10 millimolar Cu(2+) or 10 millimolar Cu(2+) + 10 millimolar Ca(2+), C(2)H(4) production increased 20- and 40-fold, respectively, within 6 hours. This increase in C(2)H(4) production was preceded and paralleled by an increase in 1-ami...
Chloroplasts can be obtained by gentle lysis or mild shear of spheroplasts of vitamin B(12)-deficient Euglena gracilis and then purified by isopycnic sedimentation on gradients of Ludox AM or Percoll. The chloroplasts appear compact and highly refractile by phase contrast or Hoffmann contrast microscopy. Upon incubatio...
Chloroplast replication in Euglena gracilis is specifically inhibited by ultraviolet light and the effect is photoreactivable.The ability of irradiated cells to be photoreactivated is lost more rapidly if cells are incubated in red light than in darkness. A mutant, Y(9)ZNa1L, which lacks the red-blue photomorphogenic s...
Thiamin pyrophosphotransferase activity was present in seedling extracts from several monocot and dicot species of agronomic as well as noncultivated plants. Changes in thiamin pyrophosphotransferase activity and thiamin pyrophosphate content were followed for 6 days in soybean (Merr.) seedlings. Maximum enzyme activit...
Thiamin:ATP pyrophosphotransferase (EC2.7.6.2) activity from soybean (Merr.) seedlings grown for 48 hours was determined by measuring the rate of [2-(14)C]thiamin incorporation into thiamin pyrophosphate. With partially purified (11-fold) enzyme, optimal activity occurred between pH 7.1 and 7.3, depending on the buffer...
An enzyme system from fruits of the flax plant is described that catalyzes the synthesis of the sugar nucleotide guanosine 5'-diphosphate l-fucose from guanosine 5'-diphosphate d-mannose with the intermediate formation of guanosine 5'-diphosphate 4-keto-6-deoxy-d-mannose. Tritium from-[(3)H]H(2)O was incorporated into ...
The possibility that fusicoccin (FC) binds to plasma membrane-associated ATPases of oat (cv. Victory) roots has been examined. Specific FC-binding in vitro is localized primarily on plasma membrane-enriched fractions. This FC-binding is greatly reduced by pretreatment of the membrane vesicles at temperatures above 45 C...
A histone-type protein kinase (EC 2.7.1.37) has been partially purified (320-fold) from the crude extracts of soybean hypocotyls by means of a combination of gel filtration and anion exchange procedures. The purified enzyme fraction is devoid of the activities of phosphoprotein phosphatase (EC 3.1.3.16), histone protea...
Twenty different legume species (20 genera) were examined for alpha-galactosidase and hemagglutinin activities. Although all of the species contained enzyme activity, only 13 of 20 contained hemagglutinin activities and none displayed a hemagglutinin activity comparable to the previously described alpha-galactosidase-h...
A soybean histone-type protein kinase was used to prepare (32)P-labeled histone H1 as substrate for purification and characterization of a phosphoprotein phosphatase (EC 3.1.3.16) from soybean hypocotyls. The phosphatase has been purified 169-fold by ammonium sulfate fractionation, ethanol precipitation, and chromatogr...
The subcellular localization of two hydrolases (ribonuclease and vicilin peptidohydrolase) which are synthesized de novo in the cotyledons of mung bean seedlings was studied. Earlier experiments had shown that both enzymes accumulate in the protein bodies in the course of seedling growth. Two methods to fractionate sub...
Biochemical changes in potato leaves during cold acclimation have been examined and compared between a frost-tolerant S. acaule and a frost-susceptible S. tuberosum species. Changes were also examined in S. tuberosum, S. acaule, and S. commersonii species when they were hardened at different temperatures to varying har...
The detergent octyl-beta-d-glucopyranoside (30 millimolar in 2 millimolar Tris-maleate, pH 7.0) preferentially extracts complexes containing protein and chlorophylls a plus b (CP) from spinach, leaving a residue highly enriched in CP I (P700-chlorophyll a protein). Use of the detergent results in a relatively gentle ex...
A Cd-binding protein has been isolated from the roots of Cd-treated tomato plants cv. Rutgers. Almost all the Cd from a high-speed supernatant fraction was recovered in a 10,000-dalton fraction from a gel filtration column coincident with 250-nanometer absorbing material. DEAE-cellulose chromatography of this 10,000-da...
Mitochondria from green leaves of spinach have been prepared using a three-step procedure involving differential centrifugation, partition in an aqueous dextran polyethylene glycol two-phase system and Percoll gradient centrifugation. The mitochondrial fractions after the different steps of purification were compared. ...
MALATE OXIDATION IN PLANT MITOCHONDRIA PROCEEDS THROUGH THE ACTIVITIES OF TWO ENZYMES: a malate dehydrogenase and a NAD(+)-dependent malic enzyme. In cauliflower, mitochondria malate oxidation via malate dehydrogenase is rotenone- and cyanide-sensitive. Addition of exogenous NAD(+) stimulates the oxidation of malate vi...
The effect of temperature on alpha-amylase synthesis and secretion from barley (c.v. Himalaya) half-seeds and aleurone layers is reported. Barley half-seeds incubated at 15 C in gibberellic acid (GA) concentrations of 0.5 and 5 micromolar for 16 hours do not release alpha-amylase. Similarly, isolated aleurone layers of...
A toxin preparation from Helminthosporium maydis Race T containing several closely related molecules with apparently identical biological activities was highly active against mitochondria and protoplasts from Texas male-sterile (T) cytoplasm corn (T mitochondria and T protoplasts, respectively) but had no effect on the...
Responses of superoxide dismutase (EC 1.15.1.1) and glutathione reductase (EC 1.6.4.2) activities were evaluated in leaf tissue from intact cotton plants (Cotton Branch 1697) which were exposed to 75% O(2), 350 microliters per liter CO(2) for 48 hours. Soluble protein was extracted from O(2)-treated and control tissue,...
Energy-dispersive x-ray analysis was used to investigate the elemental storage within protein bodies, specifically the globoid crystals, in grains of wheat. Areas of the grain investigated included various parts of the embryo, the aleurone layer plus starchy endosperm near the embryo and the aleurone layer plus starchy...
Illumination (22,000 lumens per meter(2)) of etiolated maize plants for 80 hours brings about a 5-fold increase in phosphoenolpyruvate carboxylase activity per unit of protein. An increase in carboxylase protein and incorporation of [(35)S]methionine into the protein occurs simultaneously with the activity increase. In...
Intact cotyledons were taken from pea seeds at various stages during seed development and pulse-labeled with (14)C-amino acids. Salt-soluble proteins then were extracted and fractionated on Na dodecyl sulfate-polyacrylamide gels. Storage proteins in these extracts were identified by their binding to immunoaffinity colu...
Oxygen uptake and energy charge were monitored during aging of excised maize root tips and related to the soluble sugar content and exogenous sugar supply.Oxygen uptake declined immediately after excision to 50 to 30% of its initial value after 8 and 24 hours of aging at 25 C. There was also a sharp decline of the tota...
Polysomes isolated from a susceptible variety of wheat leaves (cultivar W2691) and those inoculated with the wheat stem rust fungus (f. sp. tritici, race 126-ANZ-6, 7) were incubated in a cell-free protein-synthesizing system. Under these conditions, different size classes of polypeptides, ranging in molecular weight f...
The transport of neutral amino acids into mitochondria isolated from the hypocotyl of mung bean (Roxb.) was studied by the swelling technique. Isolated mitochondria swelled when added to an isosmotic solution of proline, serine, methionine, threonine, alanine, and glycine. The swelling was stereospecific in that it was...
Chromatographic, kinetic, and regulatory properties of glutamine synthetase in rice were investigated. By DEAE-Sephacel column chromatography, two forms (glutamine synthetase 1 and glutamine synthetase 2) were identified in leaves and one form (glutamine synthetase R) was identified in roots. Purification on hydroxyapa...
A comparative study has been carried out of the growth of two lines of Datura innoxia (Mill.) cells, designated DI-6 and NR1, their resistance to chlorate, and their ability to assimilate nitrate in sterile culture. The NR1 cell line was isolated from DI-6 cultures by first growing the latter in a nitrate-based medium ...
The storage triacylglycerols of meadowfoam (Limnanthes alba) seeds are composed essentially of C(20) and C(22) fatty acids, which contain an unusual Delta5 double bond. When [1-(14)C]acetate was incubated with developing seed slices, (14)C-labeled fatty acids were synthesized with a distribution similar to the endogeno...