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Glycine-Glomus-Rhizobium Symbiosis: II. Antagonistic Effects between Mycorrhizal Colonization and Nodulation.
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Soybean (Glycine max [L.] Merr.) plants grown in pot cultures were inoculated with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae (Nicol. & Gerd.) Gerd. and Trappe and Rhizobium japonicum strain 61A118 at planting (G(1)R(1)) or at 20 days (G(20)R(20)), or with one of the endophytes after the other has colonized the host root (G(1)R(20), G(20)R(1)). Nodulated (PR(1)) and VAM (G(1)N) dipartite associations, or nonsymbiotic plants (PN) using nutrient solutions with N, P, or N + P concentrations providing endophyte-equivalent nutrient inputs were used as controls. The delayed tripartite associations received the appropriate N, P, or N + P amendment while one or both endophytes were absent during the first 20 days of growth. Prior inoculation with one endophyte significantly inhibited development of the other. Root hexose sugar concentrations were negatively correlated with VAM colonization (r = -0.89), nodule activity (r = -0.91), and root P content (r = -0.93). Nodule (r = 0.97) and root (r = 0.96) P content correlated positively with VAM colonization. Nodule weight or VAM-fungal biomass were significantly greater in associations grown with only one endophyte. Dry weights of the PN, G(1)N, PR(1), and G(20)R(20) plants were significantly greater than those of tripartite plants inoculated at planting with either or both endophytes. Interendophyte inhibition is attributed to competition for root carbohydrates, and this effect apparently also affects overall plant productivity. The objective of the study was to determine if the timing of endophyte introduction and establishment affected the development of the other symbiotic partners.
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Bethlenfalvay, G J, Brown, M S, Stafford, A E
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Potassium and sodium absorption kinetics in roots of two tomato species : lycopersicon esculentum and lycopersicon cheesmanii.
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Excised roots of the tomato species, Lycopersicon esculentum Mill. cv Walter (the commercial species) and of Lycopersicon cheesmanii ssp. minor (Hook.) C.H. Mull. (a wild species from the Galapagos Islands), were used in comparative studies of their absorption of K(+) and Na(+). Uptake of (86)Rb-labeled K(+) and (22)Na-labeled Na(+) by excised roots of ;Walter' and L. cheesmanii varied as a function of genotype and tissue pretreatment with or without K(+). Excised roots of ;Walter' consistently absorbed more (86)Rb-labeled K(+) than those of L. cheesmanii. Absorption of K(+) from solutions ranging from 0.01 to 0.2 millimolar KCl showed saturation kinetics in both K(+)-pretreated and K(+)-depleted roots of ;Walter,' and for K(+)-depleted roots of L. cheesmanii. K(+)-pretreated roots of L. cheesmanii had exceedingly low rates of K(+) uptake with strikingly different, linear kinetics. Pretreatment with K(+) caused a decrease in rates of K(+) uptake in both genotypes. Potassium depleted roots of L. cheesmanii absorbed Na(+) at a greater rate than those of ;Walter,' whereas K(+)-pretreated roots of ;Walter' absorbed Na(+) at a greater rate than those of L. cheesmanii. The results confirm and extend previous conclusions to the effect that closely related genotypes may exhibit widely different responses to the two alkali cations, K(+) and Na(+).
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Wrona, A F, Epstein, E
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Potassium transport in two tomato species : lycopersicon esculentum and lycopersicon cheesmanii.
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The commercial tomato, lycopersicon esculentum Mill. cv Walter, and its wild relative, Lycopersicon cheesmanii ssp. minor (Hook.) C.H. Mull., were grown for 30 days under controlled conditions and in solution culture varying in its content of Na(+) and K(+). Subsequently, (86)Rb-labeled K(+) uptake and distribution were studied. From all solutions, ;Walter' consistently absorbed (86)Rb-labeled K(+) at a higher rate in micromoles per gram fresh weight per 30 minutes than L. cheesmanii. L. cheesmanii distributed a greater proportion of the absorbed K(+) from its root to its shoot. When 0.6 millimolar NaNO(3) replaced 0.6 millimolar KNO(3) in the pretreatment solution, intact plants of both genotypes followed a similar pattern as when they were pretreated with K(+) only, but a greater percentage of the absorbed K(+) remained in the roots. Leaf slices of L. cheesmanii plants deprived of K(+) for 6 days showed a greater rate of K(+) uptake than did slices from ;Walter' plants pretreated the same way. Stem slices of L. cheesmanii, however, had a lower uptake rate than did those of ;Walter'. Both leaf and stem slices of ;Walter' plants, pretreated 6 days with 0.6 millimolar NaNO(3) substituting for 0.6 millimolar KNO(3) in their growth medium, had greater rates of (86)Rb-labeled K(+) uptake from 0.5 and 20 millimolar KCl solutions than did slices of L. cheesmanii. These marked differences in patterns of ion uptake and translocation indicate that these genotypes of tomato have evolved different mechanisms to deal with K(+) and Na(+) in their environments.
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Wrona, A F, Epstein, E
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Effect of foliar applications of urea on accelerated senescence of maize induced by ear removal.
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Field grown maize (Zea mays L. cv B73 x Mo17) plants, with and without ears, were sprayed with urea solutions to determine whether foliar application of N could prevent or delay the accelerated loss of reduced N from the leaf and leaf senescence induced by ear removal. Urea sprays were applied at 7, 14, and 21 days after anthesis in three separate and equal applications that provided a total of 67 kilograms N per hectare or 1 gram N per plant. Treatments were arranged in a 2 x 2 factorial in a randomized complete block with five replicates. Appropriate plant and leaf samplings and assays were made.In response to spray treatments, net increases of reduced N were detected in the whole shoot and plant parts, especially the stalk of the earless plants and grain of the eared plants. There was no effect of urea spray treatment on the normal loss of N from the leaves or rate of senescence of the eared plants or on the accelerated loss of N from the leaves or rate of senescence induced by ear removal. Grain and stover yields were unaffected by the spray treatment.Apparently the plants were unable to utilize the urea N applied to the vegetation (primarily leaves) after anthesis to enhance or extend the accumulation of dry weight by either eared or earless plants.
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Below, F E, Crafts-Brandner, S J, Hageman, R H
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Auxin regulation of a proton translocating ATPase in pea root plasma membrane vesicles.
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Pea root microsomal vesicles have been fractionated on a Dextran step gradient to give three fractions, each of which carries out ATP-dependent proton accumulation as measured by fluorescence quenching of quinacrine. The fraction at the 4/6% Dextran interface is enriched in plasma membrane, as determined by UDPG sterol glucosyltransferase and vanadate-inhibited ATPase. The vanadate-sensitive phosphohydrolase is not specific for ATP, has a K(m) of about 0.23 millimolar for MgATP, is only slightly affected by K(+) or Cl(-) and is insensitive to auxin. Proton transport, on the other hand, is more specific for ATP, enhanced by anions (NO(3) (-) > Cl(-)) and has a K(m) of about 0.7 millimolar. Auxins decrease the K(m) to about 0.35 millimolar, with no significant effect on the V(max), while antiauxins or weak acids have no such effect. It appears that auxin has the ability to alter the efficiency of the ATP-driven proton transport.
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Gabathuler, R, Cleland, R E
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Hydrolysis of Intracellular Proteins in Vacuoles Isolated from Acer pseudoplatanus L. Cells.
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Acer pseudoplatanus cell suspension cultures were used to examine the ability of vacuoles isolated from protoplasts to hydrolyze their endogenous proteins. Total cell proteins were labeled by addition of [(3)H]leucine to the culture medium. After preparation of the protoplasts, vacuoles were isolated and were shown to be essentially free from other cellular components. Up to 30% of the [(3)H]leucine-labeled newly synthesized proteins were recovered in the vacuoles. When incubated for 6 hours at 20 degrees C, the vacuoles degraded half of these proteins. The protein breakdown was temperature and pH dependent. Analysis by electrophoresis, in denaturing polyacrylamide gels, revealed that most of the vacuolar proteins were degraded. However, some vacuolar proteins were unaffected during a 6-hour incubation period. The results indicate that vacuoles are able to acquire and degrade intracellular proteins.
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Canut, H, Alibert, G, Boudet, A M
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The senescence of detached leaves of tropaeolum.
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The senescence of detached Tropaeolum majus leaves was compared with that described earlier for Avena. Tropaeolum was chosen as being not only a dicot but also as having a nearly circular leaf, thus needing only the smallest minimum of wounding, since wounding delays the loss of chlorophyll and protein in darkness. Tropaeolum resembles Avena in that closing the stomata osmotically or with ABA causes rapid senescence in light. As in Avena also, n-hexanol and alpha,alpha'-dipyridyl delay senescence in darkness but cause ;bleaching' of chlorophyll in light. Unlike Avena, however, kinetin and gibberellic acid, which delay senescence in the dark in both species, do so in Tropaeolum without causing any significant stomatal opening. The senescence of Tropaeolum leaves is actually promoted by fusicoccin, which powerfully delays senescence in Avena, although fusicoccin does cause stomatal opening in darkness in both species. Thus, many of the phenomena of senescence are alike in the monocot and dicot, but there are several significantly different responses to the senescence-modifying reagents. It is concluded that while stomatal closure accelerates senescence in both species, stomatal opening is not directly linked to the prevention of leaf senescence.
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Thimann, K V
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A Ca/H Antiport System Driven by the Proton Electrochemical Gradient of a Tonoplast H-ATPase from Oat Roots.
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Two types of ATP-dependent calcium (Ca(2+)) transport systems were detected in sealed microsomal vesicles from oat roots. Approximately 80% of the total Ca(2+) uptake was associated with vesicles of 1.11 grams per cubic centimeter and was insensitive to vanadate or azide, but inhibited by NO(3) (-). The remaining 20% was vanadate-sensitive and mostly associated with the endoplasmic reticulum, as the transport activity comigrated with an endoplasmic reticulum marker (antimycin A-insensitive NADH cytochrome c reductase), which was shifted from 1.11 to 1.20 grams per cubic centimeter by Mg(2+).Like the tonoplast H(+)-ATPase activity, vanadate-insensitive Ca(2+) accumulation was stimulated by 20 millimolar Cl(-) and inhibited by 10 micromolar 4,4'-diisothiocyano-2,2'-stilbene disulfonic acid or 50 micromolar N,N'-dicyclohexylcarbodiimide. This Ca(2+) transport system had an apparent K(m) for Mg-ATP of 0.24 millimolar similar to the tonoplast ATPase. The vanadate-insensitive Ca(2+) transport was abolished by compounds that eliminated a pH gradient and Ca(2+) dissipated a pH gradient (acid inside) generated by the tonoplast-type H(+)-ATPase. These results provide compelling evidence that a pH gradient generated by the H(+)-ATPase drives Ca(2+) accumulation into right-side-out tonoplast vesicles via a Ca(2+)/H(+) antiport. This transport system was saturable with respect to Ca(2+) (K(m) apparent = 14 micromolar). The Ca(2+)/H(+) antiport operated independently of the H(+)-ATPase since an artifically imposed pH gradient (acid inside) could also drive Ca(2+) accumulation. Ca(2+) transport by this system may be one major way in which vacuoles function in Ca(2+) homeostasis in the cytoplasm of plant cells.
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Schumaker, K S, Sze, H
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The equilibrium of the reaction catalyzed by sucrose phosphate synthase.
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The equilibrium constant for the reaction catalyzed by sucrose phosphate synthase was reported 25 years ago to be 3250 at pH 7.5. It has been redetermined and found to be about 2 in the direction of synthesis and 6 to 10 when measured in the opposite direction.
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Barber, G A
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Corn mitochondrial protein synthesis in response to heat shock.
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Corn (Zea mays L., W23(N), OH43(N), and reciprocal single cross hybrid) seedling mitochondria respond to a 10 degrees C temperature shift (27-37 degrees C) by incorporating a greater amount of [(35)S]methionine into acid-insoluble material than mitochondria incubated at the original growing temperature (27 degrees C). This increase is in part manifested in the enhanced synthesis of a 52 kilodaltons protein. At both temperatures mitochondria of two inbreds and their reciprocal hybrids synthesize normal (N) cytoplasm proteins sensitive to chloramphenicol and insensitive to cyclohexamide treatment. The 52 kilodaltons protein is found in the supernatants of pelleted (15,000g, 5 min) mitochondria after heat shock. The role of this protein in the heat shock response is discussed in light of the implication of mitochondria as the primary cellular target to temperature stress.
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Nebiolo, C M, White, E M
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A Circadian Rhythm in the Number of Daughter Cells in Synchronous Chlorella fusca var vacuolata.
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Chlorella fusca var vacuolata cells were transferred to continuous darkness or weak light (0.07 watts per square meter) (both were called waiting time, WT) after a 12-hour light and 12-hour dark schedule. A daily dilution is performed at the end of the light/dark schedule, resulting in always the same average production of 18 autospores per mother cell. After 12 and 24 hours of WT in darkness, the production of autospores in a subsequent light/dark schedule was 50 and 100%, respectively. If the WT was performed in weak light (0.07 watts per square meter) the lowest production was obtained, independently of the length of WT. However, an interruption of this weak light by dark pulses (3 hours) increased the autospore production by an amount that depends upon the phase of the circadian rhythm, and varied up to 70% of the control (WT in permanent darkness). If the WT (total darkness) was interrupted by light pulses of 0.5 hour (white, same as used for growth), a phase response curve of productivity resulted. Pulses between the 12th and 18th hour of WT in darkness gave a 3-hour delay of maximum; later on pulses shifted the maximum autospore production 3 hours ahead.
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Wu, J T, Tischner, R, Lorenzen, H
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Effects of Irradiance on Crassulacean Acid Metabolism in the Epiphyte Tillandsia usneoides L. (Bromeliaceae).
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Spanish moss (Tillandsia usneoides L.) was collected in South Carolina, maintained in a greenhouse, then exposed to five levels of photosynthetic photon flux density (PPFD) for 3 weeks. Following this treatment, plants were sampled for chlorophyll concentrations, nocturnal acid accumulations, and photosynthetic responses to subsequent exposure at a range of PPFD. No acclimation to PPFD was observed; all plants exhibited similar patterns of nocturnal CO(2) uptake and acid accumulation regardless of initial PPFD treatment. These patterns revealed that at a PPFD level of approximately 200 micromoles per square meter per second (daytime integrated PPFD of 10 moles per square meter per day), CAM saturated or, in low-PPFD plants, was optimal. The results of this study indicate that adaptation to high PPFD is not necessarily a requirement of CAM.
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Martin, C E, Eades, C A, Pitner, R A
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A group translocator for sucrose assimilation in tonoplast vesicles of sugarcane cells.
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Existence of a group translocator for sucrose transfer into vacuoles of sugarcane (Saccharum sp.) cells has been further confirmed by the use of tonoplast vesicles isolated from intact vacuoles. The group translocator depends on external UDP-Glucose (Glc) and, via a series of enzymic reactions within the tonoplast, sucrose phosphate and sucrose are deposited inside the vesicles. Fructose-6-phosphate was not required for UDP-Glc uptake, nor was it taken up. None of the other sugar phosphates tested were taken up nor were the nucleotide sugars, UDP-Galactose and ADP-Glc. The uptake of UDP-Glc was concentration-dependent with a K(m) of 1.2 millimolar and a V(max) of 83.3 nanomoles per minute per milligram protein. The optimum pH for UDP-Glc uptake was 7.0. Uptake of UDP-Glc was inhibited by para-chloromercuribenzene-sulfonic acid, UDP, and GDP; carbonyl cyanide m-chlorophenylhydrazone inhibited to a lesser extent.
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Maretzki, A, Thom, M
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Compartmentation and equilibration of abscisic Acid in isolated xanthium cells.
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The compartmentation of endogenous abscisic acid (ABA), applied (+/-)-[(3)H]ABA, and (+/-)-trans-ABA was measured in isolated mesophyll cells of the Chicago strain of Xanthium strumarium L. The release of ABA to the medium in the presence or absence of DMSO was used to determine the equilibration of ABA in the cells. It was found that a greater percentage of the (+/-)-[(3)H]ABA and the (+/-)-trans-ABA was released into the medium than of the endogenous ABA, indicating that applied ABA did not equilibrate with the endogenous material.Therefore, in further investigations only the compartmentation of endogenous ABA was studied. Endogenous ABA was released from Xanthium cells according to the pH gradients among the various cellular compartments. Thus, darkness, high external pH, KNO(2), and droughtstress all increased the efflux of ABA from the cells. Efflux of ABA from the cells in the presence of 0.6 m mannitol occurred within 30 seconds, but only 8% of the endogenous material was released during the 20 minute treatment.
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Bray, E A, Zeevaart, J A
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Relationship of Xylem Embolism to Xylem Pressure Potential, Stomatal Closure, and Shoot Morphology in the Palm Rhapis excelsa.
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Xylem failure via gas embolism (cavitation) induced by water stress was investigated in the palm Rhapis excelsa (Thumb.) Henry. Xylem embolism in excised stems and petioles was detected using measurements of xylem flow resistance: a decrease in resistance after the removal of flow-impeding embolisms by a pressure treatment indicated their previous presence in the axis. Results supported the validity of the method because increased resistance in an axis corresponded with: (a) induction of embolism by dehydration, (b) increased numbers of cavitations as detected by acoustic means, (c) presence of bubbles in xylem vessels. The method was used to determine how Rhapis accommodates embolism; results suggested four ways. (a) Embolism was relatively rare because pressure potentials reach the embolism-inducing value of about -2.90 megapascals only during prolonged drought. (b) When embolism did occur in nature, it was confined to the relatively expendable leaf xylem; the stem xylem, which is critical for shoot survival, remained fully functional. (c) Even during prolonged drought, the extent of embolism is limited by complete stomatal closure, which occurred at the xylem pressure potential of -3.20 +/- 0.18 megapascals. (d) Embolism is potentially reversible during prolonged rains, since embolisms dissolved within 5 h at a pressure potential of 0.00 megapascals (atmospheric), and xylem sap can approach this pressure during rain.
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Sperry, J S
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Behavior and viability of tobacco protoplasts in response to electrofusion parameters.
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This investigation examines responses of protoplasts in a systematic and quantitative way to the various electrical treatments used to achieve electrofusion and their individual and cumulative effect on protoplast viability. Mesophyll and cell suspension protoplasts from two species of the same genera, Nicotiana tabacum and N. rustica var brasilia were used in these experiments. Optimal frequencies for alignment of tobacco protoplasts were between 500 kilohertz and 2 megahertz at 100 volts per centimeter. Variations in frequency and voltage of the alternating current (AC) field caused predictable movements of protoplasts within an electrofusion chamber. AC frequencies below 10 hertz or above 5 megahertz significantly decreased the viability of protoplasts in the fusion chamber as estimated by fluorescein diacetate staining 1 hour after treatment. Although the direct current (DC) pulse appeared to have a slight detrimental effect on protoplast viability, this effect was not significantly different from untreated control preparations.Protoplasts from both leaf mesophyll cells and suspension cells were induced to fuse with one or more 10 to 30 microseconds DC square wave pulses of approximately 1 kilovolt per centimeter after the protoplasts had been closely appressed with an AC field.
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Saunders, J A, Roskos, L A, Mischke, S, Aly, M A, Owens, L D
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Paclobutrazol Inhibits Abscisic Acid Biosynthesis in Cercospora rosicola.
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Three plant growth regulators, paclobutrazol, ancymidol, and decylimidazole, which are putative inhibitors of gibberellin (GA) biosynthesis, were studied to determine their effect on abscisic acid (ABA) biosynthesis in the fungus Cercospora rosicola. All three compounds inhibited ABA biosynthesis, and paclobutrazol was the most effective, inhibiting ABA 33% at 0.1 micromolar concentrations. In studies using (E,E,)-[1-(14)C] farnesyl pyrophosphate, it was shown that ancymidol blocked biosynthesis prior to farnesyl pyrophosphate (FPP), whereas paclobutrazol and decylimidazole acted after FPP. The three inhibitors did not prevent 4'-oxidation of (2Z,4E)-alpha-ionylideneacetic acid. C. rosiciola metabolized ancymidol by demethylation to alpha-cyclopropyl-alpha-(p-hydroxyphenyl)-5-pyrimidine methyl alcohol. Paclobutrazol was not metabolized by the fungus. Information that these plant growth regulators inhibit ABA as well as GA biosynthesis should prove useful in determining the full range of action of these compounds.
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Norman, S M, Bennett, R D, Poling, S M, Maier, V P, Nelson, M D
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Localization of carbamoylphosphate synthetase and aspartate carbamoyltransferase in chloroplasts.
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The localization of carbamoylphosphate synthetase (CPSase) and aspartate carbamoyltransferase (ACTase), the first two enzymes of the pyrimidine biosynthetic pathway, in chloroplasts was investigated. In dark-grown radish (Raphanus sativus) seedlings, light induced a prominent increase in CPSase activity, but had little effect on ACTase activity. Both enzymes were found in chloroplasts isolated from radish cotyledons and leaves of spinach (Spinacia oleracea), soybean (Glycine max), and corn (Zea mays). The higher activity of ACTase relative to CPSase is discussed in relation to the instability of carbamoylphosphate, the product of the CPSase, and to the control of pyrimidine synthesis. Based on these results, the function of CPSase and ACTase in chloroplasts is discussed.
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Shibata, H, Ochiai, H, Sawa, Y, Miyoshi, S
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Properties of Plasma Membrane Isolated from Chilling-Sensitive Etiolated Seedlings of Vigna radiata L.
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Plasma membrane was isolated in a uniform population and with a high purity from chilling-sensitive etiolated young seedlings of Vigna radiata (mung bean) utilizing an aqueous two polymer phase separation system and subsequent sucrose density gradient. The isolated plasma membrane was associated with vanadate-sensitive and KNO(3)-insensitive ATPase. The ATPase has high specificities both for substrate and Mg(2+) ion with optimum pH at 6.5. It was slightly stimulated by monovalent anions, especially Cl(-). Proton ionophores such as gramicidin D and carbonyl cyanide p-trifluoromethoxyphenylhydrazone did not stimulate the enzyme activity. The ATPase is apparently latent and highly stimulated by the addition of detergents such as Triton X-100. A maximum stimulation was achieved by the addition of 0.02% Triton X-100. After treatment with proteinase K in an isotonic buffer solution, the enzyme activity was less affected, whereas the peptides were specifically digested. Based on these facts, the isolated plasma membrane vesicles appear to be tightly sealed and in a right-side-out orientation. The plasma membrane ATPase had two inflection points at higher (18.9 degrees C) and lower (6.7 degrees C) temperatures on the Arrhenius plots of the activity. The lower inflection temperature apparently coincided with that of the anisotropy parameter of embedded 1,6-diphenyl-1,3,5-hexatriene, indicating that the membrane bound ATPase activity was affected by a phase transition of membrane lipids and/or temperature-dependent conformational changes in the enzyme molecules per se. Considering the fact that the plant material used here is highly sensitive to chilling temperatures and injured severely by exposure to temperatures below 5 degrees C for a relatively short period, the thermotropic properties of membrane molecules are considered to be involved in the mechanism of chilling injury.
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Yoshida, S, Kawata, T, Uemura, M, Niki, T
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Characterization of 12-oxo-phytodienoic Acid reductase in corn: the jasmonic Acid pathway.
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12-Oxo-phytodienoic acid reductase, an enzyme of the biosynthetic pathway that converts linolenic acid to jasmonic acid, has been characterized from the kernel and seedlings of corn (Zea mays L.). The molecular weight of the enzyme, estimated by gel filtration, was 54,000. Optimum enzyme activity was observed over a broad pH range, from pH 6.8 to 9.0. The enzyme had a K(m) of 190 micromolar for its substrate, 12-oxo-phytodienoic acid. The preferred reductant was NADPH, for which the enzyme exhibited a K(m) of 13 micromolar, compared with 4.2 millimolar for NADH. Reductase activity was low in the corn kernel but increased five-fold by the fifth day after germination and then gradually declined.
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Vick, B A, Zimmerman, D C
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Relationship between Photosynthesis and Protein Synthesis in Maize: I. Kinetics of Translocation of the Photoassimilated Carbon from the Ear Leaf to the Seed.
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To gain a better understanding of the biochemical basis for partitioning of photosynthetically fixed carbon between leaf and grain, a (14)CO(2) labeling study was conducted with field-grown maize plants 4 weeks after flowering. The carbon flow was monitored by separation and identification of (14)C assimilates and (14)C storage components within each tissue during the chase period (from 4 to 96 hours) following a 5 minute (14)CO(2) pulse. In the labeled ear leaf, the radioactivity strongly decreased to reach, at the end of the experiment, about 12% of the total incorporated radioactivity, mostly associated with sucrose and proteins. Nevertheless, an unexpected reincorporation of radioactivity was observed either in leaf starch or proteins, the day following the pulse. Conversely, the radioactivity in the grain increased to attain 66% of the total incorporated (14)C after a 96 hour chase. The photosynthates, mostly sucrose, organic and free amino acids, rapidly translocated towards the developing seeds, served as precursors for the synthesis of seed storage compounds, starch, and proteins. They accumulate in free form for 24 hours before being incorporated within polymerized storage components. This delay is interpreted as a necessary prerequisite for interconversions prior to the polycondensations. In the grain, the labeling of the storage molecules, either in starch or in storage protein groups (salt-soluble proteins, zein, and glutelin subgroups), was independent of their chemical nature but dependent on their pool size.
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Moutot, F, Huet, J C, Morot-Gaudry, J F, Pernollet, J C
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Ethylene-induced increase in fructose-2,6-bisphosphate in plant storage tissues.
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TREATMENT OF CARROT ROOTS WITH ETHYLENE LED TO: (a) a doubling of the fructose-2,6-bisphosphate content; (b) a general increase in the concentration of glycolytic intermediates; and (c) an increase in the extractable activity of fructose-6-phosphate,2-kinase, the enzyme synthesizing fructose-2,6-bisphosphate from fructose-6-phosphate and adenosine triphosphate.
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Stitt, M, Cséke, C, Buchanan, B
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Distinction between Cytosol and Chloroplast Fructose-Bisphosphate Aldolases from Pea, Wheat, and Corn Leaves.
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A reinvestigation of cytosol and chloroplast fructose-1,6-bisphosphate (FBP) aldolases from pea (Pisum sativum L.), wheat (Triticum aestivum L.) and corn leaves (Zea mays L.) revealed that the two isoenzymes can be separated by chromatography on diethylaminoethyl (DEAE)-cellulose although the separation was often less clear-cut than for the two aldolases from spinach leaves. Definite distinction was achieved by immunoprecipitation of the two isoenzymes with antisera raised against the respective isoenzymes from spinach leaves. The proportion of cytosol aldolase as part of total aldolase activity was 8, 9, 14, and 4.5% in spinach (Spinacia oleracea L.), pea, wheat, and corn leaves, respectively. For corn leaves we also obtained values of up to 15%. The K(m) (FBP) values were about 5-fold lower for the cytosol (1.1-2.3 micromolar concentration) than for the chloroplast enzymes (8.0-10.5 micromolar concentration). The respective K(m) (fructose-1-phosphate, F1P) values were about equal for the cytosol (1.0-2.3 millimolar concentration) and for the chloroplast aldolase (0.6-1.7 millimolar concentration). The ratio V (FIP)/V (FBP) was 0.20 to 0.27 for the cytosol and 0.07 to 0.145 for the chloroplast aldolase. Thus, cytosol and chloroplast aldolases from spinach, pea, wheat, and corn leaves differ quite considerably in the elution pattern from DEAE-cellulose, in immunoprecipitability with antisera against the respective isoenzymes from spinach leaves, and in the affinity to FBP.
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Schnarrenberger, C, Krüger, I
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Water stress enhances expression of an alpha-amylase gene in barley leaves.
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The amylases of the second leaves of barley seedlings (Hordeum vulgare L. cv Betzes) were resolved into eight isozymes by isoelectric focusing, seven of which were beta-amylase and the other, alpha-amylase. The alpha-amylase had the same isoelectric point as one of the gibberellin-induced alpha-amylase isozymes in the aleurone layer. This and other enzyme characteristics indicated that the leaf isozyme corresponded to the type A aleurone alpha-amylase (low pI group). Crossing experiments indicated that leaf and type A aleurone isozymes resulted from expression of the same genes.In unwatered seedlings, leaf alpha-amylase increased as leaf water potential decreased and ABA increased. Water stress had no effect on beta-amylase. alpha-Amylase occurred uniformly along the length of the leaf but beta-amylase was concentrated in the basal half of the leaf. Cell fractionation studies indicated that none of the leaf alpha-amylase occurred inside chloroplasts.Leaf radiolabeling experiments followed by extraction of alpha-amylase by affinity chromatography and immunoprecipitation showed that increase of alpha-amylase activity involved synthesis of the enzyme. However, water stress caused no major change in total protein synthesis. Hybridization of a radiolabeled alpha-amylase-related cDNA clone to size fractionated RNA showed that water-stressed leaves contained much more alpha-amylase mRNA than unstressed plants. The results of these and other studies indicate that regulation of gene expression may be a component in water-stress induced metabolic changes.
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Jacobsen, J V, Hanson, A D, Chandler, P C
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Chilling Sensitivity in Oryza sativa: The Role of Protein Phosphorylation in Protection against Photoinhibition.
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The effects of exposure to low temperature on photosynthesis and protein phosphorylation in chilling-sensitive and cold-tolerant plant species were compared. Chilling temperatures resulted in light-dependent loss of photosynthetic electron transport in chilling-sensitive rice (Oryza sativa L.) but not in cold-tolerant barley (Hordeum vulgare L.). Brief exposure to chilling temperatures (0-15 degrees C, 10 min) did not cause a significant difference in photosynthetic O(2) evolution capacity in vivo between rice and barley. Analysis of in vivo chlorophyll fluorescence in chilling-sensitive rice suggests that low temperatures cause an increased reduction of the plastoquinone pool that could result in photoinhibitory damage to the photosystem II reaction centers. Analysis of (32)P incorporation into thylakoid proteins both in vivo and in vitro demonstrated that chilling temperature inhibited protein phosphorylation in rice, but not in barley. Low temperature (77 K) fluorescence analysis of isolated thylakoid membranes indicated that state I to state II transitions occurred in barley, but not in rice subjected to chilling temperatures. These observations suggest that protein phosphorylation may play an important role in protection against photoinhibition caused by exposure to chilling temperatures.
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Moll, B A, Steinback, K E
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Temperature response of plasma membranes in tuber-bearing solanum species.
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Permeability coefficients (Kp) of nonelectroytes were determined at several temperatures for nonacclimated and acclimated plasma membranes of the frost sensitive Solanum tuberosum and the frost resistant Solanum commersonii. In nonacclimated membranes, Kp were equal at 25 degrees C for the two species. Kp decreased with decreased temperature in both species; however, the decrease was much greater in the frost sensitive S. tuberosum than in frost resistant S. commersonii.Kp increased with cold acclimation. After acclimation the temperature sensitivity of Kp or activation energy (Ea) for S. tuberosum was similar to that of S. commersonii; however, Kp for S. tuberosum were lower than those for S. commersonii at all temperatures.The differences in Kp and activation energy indicate plasma membrane differences between the species before acclimation. After acclimation there was still a difference between the plasma membranes of the two species as well as a difference between the nonacclimated and acclimated membranes of the same species.
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Fennell, A, Li, P H
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Studies on the Mechanism of Regulation of the mRNA Level for a Soybean Storage Protein Subunit by Exogenous l-Methionine.
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In previous studies (GL Creason et al. 1983 Biochem Biophys Res Commun 117: 658-662; LP Holowach et al. 1984 Plant Physiol 74: 576-583), we have shown that when soybean (Glycine max L. Merrill cv Provar) cotyledons are cultured in medium supplemented with l-methionine, the beta-subunit of 7S protein and beta-mRNA are absent. We have carried out further studies on the mechanism of the methionine action. In one experiment, cotyledons were cultured for 16 days with or without methionine. After 4 days, some cotyledons were transferred from methionine-supplemented to basal (no methionine) medium and vice versa. In basal medium, beta-subunit was detected at 4 days whereas in methionine-supplemented medium, no beta-subunit was present. When cotyledons were transferred from basal to methionine-supplemented medium, the beta-subunit increased within a 4 day period and then remained constant (on a per cotyledon basis). This result indicated that methionine was not acting by accelerating the degradation of the beta-subunit. Four days after transfer from supplemented to basal medium cotyledons contained beta-subunit, thus demonstrating that the inhibition was reversible. During this time, the uncombined methionine declined from 7 to 1.5 mumoles methionine per gram fresh weight. When beta-mRNA was measured by in vitro translation, functional beta-mRNA was absent in tissue that was not accumulating beta-subunit. The messenger RNA for the beta-subunit had a half-life of about 1 day in the presence of methionine. Hybridization of cotyledon mRNA with cDNA complementary to beta-mRNA revealed that the 1700 nucleotide beta-mRNA was not present in supplemented cotyledons. Thus, expression of the beta-subunit gene is controlled at the level of transcription, RNA processing, or RNA turnover, rather than at the level of translation.
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Holowach, L P, Madison, J T, Thompson, J F
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Plastid DNA content in a cultured soybean line capable of photoautotrophic growth.
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The levels of chloroplast DNA in a cultured photoautotrophic soybean (Glycine max [L.] Merr. v Corsoy) cell line were determined by molecular hybridization. The cells were also grown photomixotrophically and heterotrophically as suspension cultures and the level of plastid DNA was found to be constant at approximately 26% of the total cellular DNA in all three growth modes. By comparison, total cellular DNA extracted from plants of the same variety used as the explant source for the cultured cells contained 12.3 to 18.9% (leaves and seeds) and 6.1 to 8.9% (roots) plastid DNA.
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Cannon, G, Heinhorst, S, Weissbach, A
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Sugar utilization by developing wild type and shrunken-2 maize kernels.
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To characterize the movement of sugars during kernel development in maize, a newly devised in vitro kernel development scheme was utilized. Viable seeds of wild type maize (Zea mays L.) as well as the mutant shrunken-2 (sh2) were found to mature when grown in culture with reducing sugars or sucrose as the carbon source. However, wild type and sh2 kernels had greater germination, starch content, and seed weight when sucrose, rather than reducing sugars, was the carbon source. By the use of labeled sucrose it was shown that sucrose can move into endosperm tissue without intervening degradation and resynthesis. These results show that when grown in vitro the maize seed can utilize reducing sugars for development, but it prefers sucrose.
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Cobb, B G, Hannah, L C
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District programme to reduce smoking: can sustained intervention by general practitioners affect prevalence?
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A total of 101 general practitioners in 27 practices in inner London took part in a quasi-experimental study designed to examine whether a brief intervention applied to all smokers seen by general practitioners and sustained on a continuous basis could in time have a cumulative effect and reduce the prevalence of smoking among their patients. Of 21 practices approached in our local district (Camberwell), seven were willing to undertake brief intervention with support from the smokers' clinic (SBI), four opted for intervention without support (BI), and six acted as usual care controls. A further 10 out of 12 practices approached in South Hammersmith provided an unselected group of usual care controls. A series of six cross-sectional surveys were conducted over a three-year period. Each survey consisted of all adult patients attending to see a doctor during a defined two-week period, sample sizes averaging just over 9000 per survey. The estimated decline in self-reported smoking prevalence over the 30-month period following the start of intervention was 5.5% (from 36.4% to 30.9%) in the SBI group compared with 2.1% for BI and 2.8% and 3.0% in the two usual care control groups, the decline in the SBI group being significantly greater than in the other groups which did not differ significantly between each other. These interim results provide encouraging evidence that brief intervention by general practitioners with support and back-up from a local smokers' clinic can, when sustained on a continuous basis, reach sufficient smokers to reduce smoking prevalence in their practice populations. However, firm conclusions must await longer periods of observation now that the other Camberwell practices have adopted the SBI procedures.
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Russell, M A, Stapleton, J A, Hajek, P, Jackson, P H, Belcher, M
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Levels of customary physical activity among the old and the very old living at home.
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With an activity inventory designed specifically for use among elderly people, detailed profiles of customary physical activity were obtained from 507 old (aged 65-74 years) and 535 very old (aged 75 years and over) individuals randomly sampled from the community. Participation in four categories of activity was assessed: outdoor productive activities; indoor productive activities; leisure activities; and walking. Customary engagement in many activities was found to be low, age (old versus very old) and sex being among the most important determinants of participation. The method of assessment is described, and activity profiles normative for older age groups are presented.
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Dallosso, H M, Morgan, K, Bassey, E J, Ebrahim, S B, Fentem, P H, Arie, T H
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High and low risk groups for cancer of colon and rectum in Denmark: multiplicative Poisson models applied to register linkage data.
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Multiplicative Poisson models were used to identify subgroups of the Danish population at high and low risk of developing cancer of the right or left side of the colon, and cancer of the rectum. The analysis was based on cross-linked data from the 1970 census and the Danish Cancer Registry, where a 10-year follow-up period yielded some 20,000 colo-rectal cancers, in approximately 2.5 million persons. The risk of cancer of the right side of the colon in longer educated men living in apartment houses was almost twice as high as in farmers living in single family houses (relative risk 1.84; 95% confidence interval 1.42-2.37). A two-fold ratio (RR 2.18; 95% CI 1.70-2.62) was also seen in the risk of cancer of the left side of the colon between men with longer education in Greater Copenhagen and farmers in Jutland. The annual number of colon cancer cases in men in Denmark could be reduced by 27% if the incidence for all men was equal to that found for the low risk group of farmers.
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Mellemgaard, A, Engholm, G, Lynge, E
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Gastro-intestinal atresias in Finland in 1970-79, indicating time-place clustering.
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Information on oesophageal, duodenal and rectal atresias was collected from the hospitals, malformation register and death certificates in Finland from 1970-79. The respective prevalence rates were 4.1, 1.4 and 3.6 per 10,000 births, which are among the highest rates cited in the literature. Mothers of children with oesophageal and duodenal atresias were older than the average for child-bearing women in Finland. The sex ratio (male/female) of rectal atresia patients was 2.4. About 50% of the atresia patients had associated malformations. The data indicated time-spaced clustering for atresias. Furthermore, the cases of oesophageal and duodenal atresias appeared to spread within a time period of 30 days from a province to its neighbouring provinces to the north. Spreading was not observed for rectal atresias but clustering was noted when smaller geographic areas and individual communities were analysed. Occupations involving animal contacts were overpresented among the fathers of atresia patients. The data suggest infective aetiology, probably with no known clinical symptoms and no seasonal patterns.
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Kyyrönen, P, Hemminki, K
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Epidemiological aspects of suicide among the young in selected European countries.
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