Biomedical subjects

H Kende

Publications and source records attributed to H Kende.

At least 55 records · Page 3Linked to original sources

PubMed1985

Ethylene and the growth of rice seedlings.

Etiolated whole rice seedlings enclosed in sealed vials produced ethylene at a rate of 0.9 picomole per hour per seedling. When 2-centimeter-long shoots were subdivided into 5-millimeter-long sections, the sections containing the tip of the shoot evolved 37% of the total ethylene with the remaining 63% being produced along a gradient decreasing to the base of the shoot. The tip of the coleoptile also had the highest level of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid and of the ethylene-forming enzyme activity. Ethylene is one of the factors controlling coleoptile elongation. Decapitation of the seedling reduced ethylene evolution to one-third its original level and inhibited coleoptile growth. In short-term experiments, the growth rate of decapitated seedlings was restored to almost that of intact seedlings by application of ethylene at a concentration of 10 microliters per liter. Apart from ethylene, O(2) also participates in the control of coleoptile growth. When rice seedlings were grown in a gas mixture of N(2) and O(2), the length of the coleoptiles reached a maximum at a concentration of 2.5% O(2). Lower and higher concentrations of O(2) reduced coleoptile growth. The effect of exogenous ethylene on coleoptile growth was also O(2) dependent.

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PubMed1984

Effect of submergence on the cell wall composition of deep-water rice internodes.

The cell wall composition of internodes of deep-water rice plants (Oryza sativa L. cv Habiganj Aman II) which were induced to grow rapidly by submergence in water was compared to that of nonsubmerged plants which grew slowly. No differences could be detected in cellulose, uronic acid, and lignin content expressed on a dry weight basis. Cell wall preparations of rapidly growing, submerged internodes contained more hydroxyproline and had a higher hydration capacity than those of control internodes. The silicon content of submerged rice internodes was considerably lower than that of air-grown plants. The role of silicon as a structural component of the cell wall of grasses is discussed in relation to lodging of deep-water rice plants after the flood waters have receded.

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PubMed1984

Role of gibberellin in the growth response of submerged deep water rice.

We have shown previously that ethylene, which accumulates in the air spaces of submerged stem sections of rice (Oryza sativa L. cv "Habiganj Aman II"), is involved in regulating the growth response caused by submergence. The role of gibberellins in the submergence response was studied using tetcyclacis (TCY), a new plant growth retardant, which inhibits gibberellin biosynthesis. Stem sections excised from plants that had been watered with a solution of 1 micromolar TCY for 7 to 10 days did not elongate when submerged in the same solution or when exposed to 1 microliter per liter ethylene in air. Gibberellic acid (GA(3)) at 0.3 micromolar overcame the effect of TCY and restored the rapid internodal elongation in submerged and ethylene-treated sections to the levels observed in control sections that had not been treated with TCY. The effect of 0.01 to 0.2 micromolar GA(3) on internodal elongation was enhanced two- to eight-fold when 1 microliter per liter ethylene was added to the air passing through the chamber in which the sections were incubated. GA(3) and ethylene caused a similar increase in cell division and cell elongation in rice internodes. Thus, ethylene may cause internodal elongation in rice by increasing the activity of endogenous GAs. In internodes from which the leaf sheath had been peeled off, growth in response to submergence, ethylene and GA(3) was severely inhibited by light.

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PubMed1983

Properties and Partial Purification of 1-Aminocyclopropane-1-carboxylate Synthase.

We studied the regulation of 1-aminocyclopropane-1-carboxylate (ACC) synthase activity in tomato (Lycopersicon esculentum Mill.) fruit tissue and attempted the purification of this enzyme. The increase of ACC synthase activity in wounded tomato pericarp was inhibited by cordycepin and cycloheximide. Density labeling studies showed a 0.75% increase in the buoyant density of ACC synthase isolated from tomato pericarp tissue that had been incubated on (2)H(2)O as compared to ACC synthase from H(2)O-treated tissue. These data are consistent with the hypothesis that ACC synthase is synthesized de novo following wounding of tomato pericarp tissue. SDS-gel electrophoresis and fluorography showed that the pattern of incorporation of l-[(35)S]methionine into protein changed with time after wounding of the tissue. Radioactive protein bands that were not detected 1 hour after wounding, became apparent 2 to 3 hours after wounding.Gel filtration on Sephadex G-100 gave a molecular weight estimate for ACC synthase of 57,000 +/- 1,500 daltons. Hydrophobic interaction chromatography on phenyl-Sepharose yielded a 60- to 70-fold purification of the enzyme. SDS-gel electrophoresis of this preparation indicated the presence of one intense band at 57,000 daltons and several less intense bands. Affinity chromatography was of limited usefulness in the purification of ACC synthase since the enzyme could not be eluted specifically from any of the affinity gels tried. Purification methods that involved pH changes led to a rapid loss of ACC synthase activity. ACC synthase was estimated to comprise less than 1% of the total protein in tomato pericarp tissue.

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PubMed1983

The role of ethylene in the growth response of submerged deep water rice.

We investigated the effect of partial submergence on internode elongation in a Bangladesh variety of floating or deep water rice (Oryza sativa L., cv. Habiganj Aman II). In plants which were at least 21 days old, 7 days of submergence led to a 3- to 5-fold increase in internodal length. During submergence, the ethylene concentration in the internodes increased from about 0.02 to 1 microliters per liter. Treatment of nonsubmerged plants with ethylene also stimulated internode elongation. When ethylene synthesis in partially submerged plants was blocked with aminooxyacetic acid and aminoethoxyvinylglycine, internode elongation was inhibited. This growth inhibition was reversed when ethylene biosynthesis was restored with 1-aminocyclopropane-1-carboxylic acid (ACC). Radio-labeling studies showed that ethylene in floating rice was synthesized from methionine via ACC. Internodal tissue from submerged plants had a much higher capacity to form ethylene than did internodal tissue from nonsubmerged plants. This increase in ethylene synthesis appeared to be due to enhanced ACC formation rather than to increased conversion of ACC to ethylene. Our results indicate that ethylene produced during submergence is required for the stimulation of growth in submerged floating rice plants.

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PubMed1983

How does deep water rice solve its aeration problem.

In partially flooded deep water rice (Oryza sativa L. cv Habiganj Aman II), continuous air layers trapped between the hydrophobic, corrugated surface of the leaf blades and the surrounding water constitute the major path of aeration. The conduction of gases through the internal air spaces of the leaf is negligible compared to the conduction of gases through the external air layers. The total volume of the air layers on both sides of a leaf blade is about 45% of the volume of the leaf blade itself. The size of the air layers around submerged leaf blades of cereals not adapted to conditions of partial flooding, e.g. of oats, barley, and wheat, is considerably smaller than that of rice. Gases move through the air layers not only by diffusion but also by mass flow. In darkness, air is drawn down from the atmosphere through the air layers along a pressure gradient created by solubilization of respiratory CO(2) in the surrounding water. In light, photosynthetic O(2) is expelled through the air layers to the atmosphere because the solubility of O(2) in water is much lower than that of CO(2). Air layers greatly increase the rate of photosynthetic carbon fixation by enlarging the surface of the gas-liquid interface available for CO(2) uptake from the water. Air layers are vital for the survival of the partially submerged rice plant. When leaves are washed with a dilute solution of a surfactant (Triton X-100), no air layers are formed under water. Plants without air layers do not grow in response to submergence, and the submerged parts of the plant deteriorate as evident by rapid loss of chlorophyll and protein. Air layers provide a significant survival advantage even to completely submerged rice plants.

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PubMed1980

Ethylene Action and Loss of Membrane Integrity during Petal Senescence in Tradescantia.

Senescence of isolated petals of Tradescantia is accompanied by a large increase in membrane permeability, and application of ethylene hastens the onset of this increase. There is a 1- to 2.5-hour lag between ethylene application and the onset of anthocyanin efflux (an indicator of increased membrane permeability). Simultaneous application of 0.1 millimolar cordycepin or cycloheximide with ethylene abolishes the response to ethylene. Analysis of phospholipid levels in these petals during senescence has shown that the increase in membrane permeability is accompanied by a massive loss of phospholipids. Factors which enhance or retard the rate of anthocyanin efflux exert a corresponding effect on the rate of phospholipid loss. The composition of the phospholipid fraction remains unchanged during senescence. The activity of phospholipase D declines during senescence whereas that of acyl hydrolase remains essentially constant.

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PubMed1980

Effect of 1-Aminocyclopropane-1-Carboxylic Acid on the Production of Ethylene in Senescing Flowers of Ipomoea tricolor Cav.

Application of 1-aminocyclopropane-1-carboxylic acid (ACC) to rib segments excised from flowers of Ipomoea tricolor Cav. resulted in the formation of C(2)H(4) in greater quantities than produced under natural conditions. The ability of ACC to enhance C(2)H(4) production was independent of the physiological age of the tissue and its capacity to synthesize C(2)H(4) without applied ACC. When ACC was fed to rib segments that had been treated with [(14)C]methionine, incorporation of radioactivity into C(2)H(4) was reduced by 80%. Aminoethoxyvinylglycine and aminooxyacetic acid inhibited C(2)H(4) production in rib segments of I. tricolor but had no effect on ACC-enhanced C(2)H(4) production. Protoplasts obtained from flower tissue of I. tricolor did not form C(2)H(4), even when incubated with methionine or selenomethionine. They produced C(2)H(4) upon incubation with ACC, however. ACC-dependent C(2)H(4) production in protoplasts was inhibited by n-propyl gallate, AgCl, CoCl(2), KCN, Na(2)S, and NaN(3). ACC-dependent C(2)H(4) synthesis in rib segments and protoplasts was dependent on O(2), the K(m) for O(2) being 1.0 to 1.4% (v/v). These results confirm the following pathway for C(2)H(4) biosynthesis in I. tricolor. methionine [selenomethionine] --> S-adenosylmethionine [selenoadenosylmethionine] --> ACC --> C(2)H(4).

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PubMed1979

Interactions of Methionine and Selenomethionine with Methionine Adenosyltransferase and Ethylene-generating Systems.

Since selenomethionine appears to be a better precursor of ethylene in senescing flower tissue of Ipomoea tricolor and in indole acetic acid-treated pea stem sections than is methionine (Konze JR, N Schilling, H Kende 1978 Plant Physiol 62: 397-401), we compared the effectiveness of selenomethionine and methionine to participate in reactions which may be connected to ethylene biosynthesis. Evidence is presented that selenomethionine is also a better substrate of methionine adenosyltransferase (ATP: methionine S-adenosyltransferase, EC 2.5.1.6) from I. tricolor, the V(max) for selenomethionine being twice as high as that for methionine. The affinity of the enzyme is higher for methionine than for selenomethionine, however. Methionine added to flower tissue together with selenomethionine inhibits the enhancement of ethylene synthesis by the seleno analog. Likewise, methionine reduces the high, selenomethionine-dependent reaction rates of methionine adenosyltransferase from I. tricolor flower tissue. On the other hand, selenomethionine is less effective as an ethylene precursor than is methionine in model systems involving oxidation by free radicals. It was concluded that activation of methionine by methionine adenosyltransferase and formation of S-adenosylmethionine are more likely to be involved in ethylene biosynthesis than is oxidation of methionine by free radicals.

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PubMed1979

Methionine metabolism and ethylene formation in etiolated pea stem sections.

Stem sections of etiolated pea seedlings (Pisum sativum L. cv. Alaska) were incubated overnight on tracer amounts of l-[U-(14)C]methionine and, on the following morning, on 0.1 millimolar indoleacetic acid to induce ethylene formation. Following the overnight incubation, over 70% of the radioactivity in the soluble fraction was shown to be associated with S-methylmethionine (SMM). The specific radioactivity of the ethylene evolved closely paralleled that of carbon atoms 3 and 4 of methionine extracted from the tissue and was always higher than that determined for carbon atoms 3 and 4 of extracted SMM.Overnight incubation of pea stem sections on 1 millimolar methionine enhanced indoleacetic acid-induced ethylene formation by 5 to 10%. Under the same conditions, 1 millimolar homocysteine thiolactone increased ethylene synthesis by 20 to 25%, while SMM within a concentration range of 0.1 to 10 millimolar did not influence ethylene production. When unlabeled methionine or homocysteine thiolactone was applied to stem sections which had been incubated overnight in l-[U-(14)C]methionine, the specific radioactivity of the ethylene evolved was considerably lowered. Application of unlabeled SMM reduced the specific radioactivity of ethylene only slightly.

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PubMed1979

Hydrolytic enzymes in the central vacuole of plant cells.

The hydrolase content of vacuoles isolated from protoplasts of suspension-cultured tobacco cells, of tulip petals, and of pineapple leaves, and the sedimentation behavior of tobacco tonoplasts were studied. Three precautions were found to be important for the analysis of vacuolar hydrolases and of the tonoplast. (a) Purification of protoplasts in a Ficoll gradient was necessary to remove cell debris which contained contaminating hydrolases adsorbed from the fungal cell-wall-degrading enzyme preparation. (b) Hydrolase activities in the homogenates of the intact cells or the tissue used and of the purified protoplasts had to be compared to verify the absence of contaminating hydrolases in the protoplast preparation. (c) Vacuoles obtained from the protoplasts by an osmotic shock had to be purified from the lysate in a Ficoll gradient. Since the density of the central vacuole approximates that of the protoplasts, about a 10% contamination of the vacuolar preparation by surviving protoplasts could not be eliminated and had to be taken into account when the distribution of enzymes and of radioactivity was calculated.THE INTRACELLULAR ACTIVITIES OF THE FOLLOWING ACID HYDROLASES WERE PRIMARILY LOCALIZED IN THE VACUOLE OF TOBACCO CELLS: alpha-mannosidase, beta-N-acetylglucosaminidase, beta-fructosidase, nuclease, phosphatase, phosphodiesterase. A similar composition of acid hydrolases was found in vacuoles obtained from protoplasts of tulip petals. Proteinase, a hydrolase with low activity in tobacco cells and tulip petals and therefore difficult to localize unequivocally, was found to be vacuolar in pineapple leaves, a tissue containing high levels of this enzyme. Our data support the hypothesis that the central vacuole of higher plant cells has an enzyme composition analogous to that of the animal lysosome.None of the vacuolar enzymes investigated was found to be bound to the tonoplast. When vacuoles were isolated from cells labeled with radioactive choline, the vacuolar membrane was found to contain radioactivity. On sucrose gradients, the label incorporated into tonoplasts banded around a density of 1.10 grams per cubic centimeter (24% sucrose, w/w).

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PubMed1978

No Effect of 5-Fluorouracil on the Properties of Purified alpha-Amylase from Barley Half-seeds.

alpha-Amylase has been purified from de-embryonated seeds of barley (Hordeum vulgare L. cv. Betzes) which have been incubated on 10(-6)m gibberellic acid (GA(3)) following 3 days of imbibition in buffer. Incubation of the half-seeds in up to 10(-2)m 5-fluorouracil (5-FU) during the entire incubation period, including imbibition, had no effect on any of the following characteristics of purified alpha-amylase: thermal stability in the absence of calcium, molecular weight of the enzyme, isozyme composition, specific activity, or the amount of alpha-amylase synthesized by the aleurone tissue. The synthesis of rRNA and tRNA was strongly inhibited by 5-FU, indicating that the analog had entered the aleurone cells. These results are not in agreement with those of Carlson (Nature New Biology 237: 39-41 [1972]) who found that treatment of barley aleurone with 10(-4)m 5-FU prior to the addition of GA(3) resulted in decreased thermal stability of GA(3)-induced alpha-amylase and who interpreted this as evidence that the mRNA for alpha-amylase was synthesized during the imbibition of the aleurone tissue and independently of gibberellin action. Results of the present experiments indicate that the thermal stability of highly purified alpha-amylase is not altered by treatment of barley half-seeds with 5-FU, and that 5-FU cannot be used as a probe to examine the timing of alpha-amylase mRNA synthesis.

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PubMed1978

Ethylene and senescence in petals of tradescantia.

Flowers of Tradescantia (clone O2) which are ephemeral, produce ethylene during senescence with the maximum rates occurring during the initial period of fading. Senescing isolated petals produce ethylene in a similar manner, exhibit a loss of membrane semipermeability, and exogenous ethylene hastens the onset as well as the subsequent rate of this loss. The aminoethoxy analog of 0.1 millimolar rhizobitoxine completely inhibits ethylene production by isolated petals but only partially the loss of membrane semipermeability. Isolated petals acquire a sensitivity to ethylene as they mature, becoming fully sensitive on the day of anthesis.

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PubMed1978

Enhancement of ethylene formation by selenoamino acids.

Selenomethionine and selenoethionine enhanced ethylene production in senescing flower tissue of Ipomoea tricolor Cav. and in auxin-treated pea (Pisum sativum L.) stem sections. This enhancement was fully inhibited by the aminoethoxy analog of rhizobitoxine. Methionine did not have a comparable promotive effect, and ethionine partly inhibited ethylene production. When [(14)C]methionine was applied to flower or pea stem tissue followed by treatment with unlabeled selenomethionine or selenoethionine, the specific radioactivity of the ethylene evolved was considerably reduced. The dilution of the specific radioactivity of ethylene by selenomethionine, and in pea stem sections also by selenoethionine, was greater than the dilution by nonradioactive methionine at the same concentration. These results indicate that both selenoamino acids serve as precursors of ethylene and that they are converted to ethylene more efficiently than is methionine.

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PubMed1977

Membrane Lipids in Senescing Flower Tissue of Ipomoea tricolor.

Rib segments excised from flower buds of Ipomoea tricolor Cav. pass through the same phases of senescence as the respective tissue on the intact plant. Such segments were used to correlate changes in lipid content with known symptoms of aging, such as rolling up of the ribs and ethylene formation. It was found that the level of phospholipid had already started to decline before visible signs of senescence were evident. As the segments began to roll up and to produce ethylene, the rate of phospholipid loss accelerated sharply. During the same period, the level of fatty acids esterified to phospholipids also fell by 40%. No qualitative changes in any lipid component could be detected during senescence. Labeling experiments using (33)P as marker showed that the rate at which radioactivity was lost from phospholipids during aging was parallel to the rate at which the level of total phospholipids declined. Exogenously applied ethylene accelerated the loss of phospholipid and the senescence of rib segments while benzyladenine retarded both of these processes.Ag(+), which counteracts the effect of ethylene in many plants, inhibited rolling up of the rib segments but did not affect either spontaneous and ethylene-induced ethylene generation, or phospholipid loss. In contrast, Co(2+), a purported inhibitor of ethylene synthesis, reduced ethylene production, rolling up, and phospholipid loss. The inhibition of ethylene-induced rolling up by Co(2+) could not be overcome with exogenous ethylene, however.Our results indicate that phospholipid loss is a marker for membrane degradation in rib segments. Changes in membrane integrity and in cellular compartmentation may be the basis for ethylene synthesis during aging of flower tissue.

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PubMed1976

Relationship between Ethylene Evolution and Senescence in Morning-Glory Flower Tissue.

An excised tissue system consisting of corolla rib segments was developed to study the relationship between senescence and ethylene production in morning-glory flowers (Ipomoea tricolor). Such segments, isolated 1 or 2 days (day -1 or day -2) before flower opening (day 0) passed through the same developmental phases as did the corresponding tissues of the intact organ. When excised on day -1 and incubated overnight, the rib segments turned from purple to blue and changed from a slightly curled to a flat configuration. On day 0, these segments rolled up during the afternoon and turned purple again, as did the ribs of an intact corolla; the rolling up coincided with an increased rate of ethylene production. Premature rolling up and associated ethylene evolution were induced by ethylene or propylene treatment. When segments were excised on day -2 and incubated overnight, there were no changes in color or shape; during day -1, no spontaneous rolling up and little ethylene evolution occurred. Application of ethylene or propylene to these immature segments elicited rolling up but did not stimulate endogenous ethylene production.Overnight treatment of segments cut on day -1 with 10(-6)m benzyladenine markedly retarded spontaneous rolling up and ethylene evolution, although the response to applied ethylene was only slightly slowed. Overnight treatment of segments cut on day -1 with the ethoxy analog of rhizobitoxine (10(-5) to 10(-4)m) resulted in almost complete (>99%) inhibition of both spontaneous and propylene-induced ethylene evolution. Although spontaneous rolling up was delayed, it was not abolished, and ethylene-induced rolling up was almost unaffected.These data indicate that an ethylene-generating system develops as an integral part of the aging process in flower tissue. Ethylene hastens aging of the flower, but may not play an obligatory role in flower senescence.

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PubMed1976

Methionine metabolism and ethylene biosynthesis in senescent flower tissue of morning-glory.

In immature rib segments prepared from morning-glory (Ipomoea tricolor) flower buds, the major soluble metabolite formed from tracer amounts of l-methionine-U-(14)C was S-methylmethionine (SMM). In segments of senescing ribs, (14)C was progressively lost from SMM and appeared in free methionine. Immature segments contained about 4 nmoles of free methionine and about 16 nmoles of SMM per 30 segments. As the segments senesced, the methionine content increased about 10-fold while the SMM content remained unchanged; during this time about 0.8 nmole of ethylene was produced per 30 segments. Tracer experiments with l-methionine-U-(14)C, l-methionine-methyl-(3)H, and l-homocysteine thiolactone-(35)S indicated that SMM was capable of acting as a methyl donor, and that in senescent segments the methyl group was utilized for methionine production with homocysteine serving as methyl acceptor. Of the 2 molecules of methionine produced in this reaction, 1 was re-methylated to SMM, and the other contributed to the observed rise in the content of free methionine.Internal pools of methionine and SMM were prelabeled (but not significantly expanded) by overnight incubation on 10 mum l-methionine-U-(14)C. The specific radioactivity of the ethylene subsequently evolved during the senescence of the segments closely paralleled the specific radioactivity of carbon atoms 3 plus 4 of free methionine extracted from the tissue, demonstrating that methionine was the major precursor of ethylene in this system. The specific radioactivity of carbon atoms 3 plus 4 of extracted SMM was about twice that of the free methionine.Based on these results, a scheme for methionine biosynthesis in senescent rib tissue is presented. The operation of this pathway in the control of ethylene production is discussed.

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PubMed1976

Biosynthesis of wound ethylene in morning-glory flower tissue.

Production of wound ethylene was investigated in rib segments excised from flower buds of morning-glory (Ipomoea tricolor). Segments of the ribs were cut from buds 2 days before flower opening, floated overnight on 5 mm KCl solution, and transferred to agar the following morning. These immature segments evolved only a small quantity of ethylene during incubation on agar, with most of the production occurring in the morning. When such segments were wounded mechanically early in the afternoon, the rate of ethylene production rose more than 10-fold within 1 hour and returned to a low rate after about 3 hours.Production of ethylene by both untreated and wounded rib segments was inhibited more than 95% by overnight pretreatment with the ethoxy analog of rhizobitoxine (3 x 10(-5) and 10(-4)m). After overnight exposure of segments to 9 muml-methionine-U-(14)C, the specific radioactivity of the ethylene evolved by untreated and wounded tissue was determined and compared to the specific radioactivities of carbon atoms 3 plus 4 of methionine and S-methylmethionine (SMM) extracted from the segments. The specific radioactivity of methionine was about one-half that of SMM; neither value was significantly affected by wounding. The specific radioactivity of ethylene evolved by untreated tissue was close to that of SMM. In wounded tissue the specific radioactivity of the ethylene evolved was lower, but still above that of methionine. These results are consistent with the interpretations that wound ethylene is synthesized from carbon atoms 3 plus 4 of either SMM or methionine. On the basis of earlier experiments with senescing rib segments, it is suggested that methionine serves as the precursor of the wound ethylene.

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