Effect of zeatin on the growth and indolyl-3-acetic acid and abscisic acid levels in maize roots

Elongation, indolyl-3-acetic acid (IAA) and abscisic acid (ABA) levels, – gas chromatography-mass spectrometry quantification –, in the elongating zone were analysed for maize ( Zea mays L., Cv. LG11) roots immersed in buffer solution with or without zeatin (Z). The effect of Z depends on the initial extension rate of roots. The slower growing roots are more strongly inhibited by Z (10⁻⁷−10₋₅ M ) and they show a greater increase in IAA and ABA content. When compared to the rapidly growing roots, the larger reactivity of the 'slow'ones cannot be attributed to a higher Z uptake as shown when using [¹⁴C]-Z. It is suggested that Z could regulate root elongation by acting on the IAA and/or ABA level. The comparative action of these two hormones is discussed.     

Adventitious root formation 'in vitro' in apple rootstocks (Malus pumila) II. Uptake and distribution of indol-3yl-acetic acid during the auxin-sensitive phase in M.9 and M.26

During the auxin-sensitive phase of root initiation, rates of 3-indolyl- [2-¹⁴C] acetic acid (IAA) uptake into the 1 cm bases of shoots of the apple rootstock M.9 ( Malus pumila Mill.) 'in vitro' were not significantly affected by the presence of 10⁻³ M phloroglucinol (PG) using either liquid or agar-solidified media. The use of a liquid medium did however reduce rates of uptake over a 10-day period of auxin application. The distribution of labelled IAA between the 1-cm base and the shoot remainder was not affected by PG.
Exposure of shoots of the difficult-to-root M.9 and the easy-to-root M.26, to 2.8 × 10⁻⁵ M IAA containing [2-¹⁴C] IAA revealed no positive correlation between the amount of label taken up by the 1-cm base and rooting performance. M.9 bases absorbed almost twice as much label as M.26 after 9 days but had produced only one-third as many roots. Measurements of label distribution between the 1-cm base and the shoot remainder showed that less than 10% of the label moved to the shoot remainder over a 6-day period of auxin application. Dose-response curves of IAA and rooting over the range 1 × 10⁻⁵ M and 3 × 10⁻³ M showed that root number in M.9 was at an optimum at 1 × 10⁻³ M IAA after 6 days whilst M.26 required only 1 × 10⁻⁴ M for a similar response. These data support the hypothesis that differences in rooting of the two rootstocks reflect differences in the endogenous metabolism of exogenous IAA and not differences in its rates of uptake or distribution in the shoots.     

Identification of jasmonic acid in Mimosa pudica and its inhibitory effect on auxin- and light-induced opening of the pulvinules

Jasmonic acid was identified from Mimosa pudica L. plants by mass spectrometry, high performance liquid chromatography and thin layer chromatography. Effects of authentic jasmonic acid on pulvinule movement and transpiration of the pinnae were compared with those of abscisic acid. Jasmonic acid and abscisic acid each at 10⁻⁵ M inhibited both auxin- and light-induced opening of the pulvinules. A closure-inducing activity of jasmonic acid at 10⁻⁴ M was greater than that of abscisic acid at 10⁻⁴ M. Pinnae transpiration was reduced by 10⁻⁵ M abscisic acid but not by 10⁻⁴ M jasmonic acid.     

β-Adrenergic antagonists and carp (Cyprinus carpio L.) sperm motility

We studied the effects of two β-adrenergic antagonists, atenolol and propranolol, on carp sperm motility. Atenolol (10⁻³−10⁻⁸ m ) has no appreciable effects while propranolol (6 × 10⁻⁵−3 × 10⁻⁶ m ) affects the percentage of sperm motility in a dose-dependent fashion.     

Effects of a brassinosteroid on growth and electrogenic proton extrusion in maize root segments

In apical or subapical root segments of maize ( Zea mays L. cv. Dekalb XL640), 10⁻⁷–10⁻⁵ M 24-epibrassinolide (BR), a physiologically active synthetic epimer of the pollen hormone brassinolide, induces a significant stimulation of root growth, associated with an increase of acid secretion. The increase in acid secretion is enhanced by the presence of K⁺ in the medium and is accompanied by an early, significant hyperpolarization of the transmembrane electric potential (PD), which is completely suppressed by the addition of the protonophore uncoupler FCCP. Similar effects of BR have earlier been reported for shoots, and also for IAA in shoots. Contrariwise, 10⁻⁸–10⁻⁷ M IAA inhibits acid secretion and depolarizes the PD in the maize root segments. This suggests different pathways for the action of the two different hormones on the proton pump.     

Bioelectrical reactions of Nitellopsis obtusa induced by indole-3-acetic acid

The complex of bioelectrical paramenters (membrane potential, membrane resistance and capacitance) of internodal cells of Nitellopsis obtusa was measured over a wide range of IAA concentration (10⁻¹⁰ to 10⁻⁴ M ) with two intracellular microelectrodes. Primary effects of IAA at a concentration as low as 10⁻¹⁰ M were observed. The optimum range of IAA action was from 10⁻⁹ to 10⁻⁶ M . The type of IAA-induced electroresponse depended on the initial level of membrane potential, which characterized the energetic state of the plasmalemma. In the energized state (ca −200 mV) N. obtusa cells appeared to have 3 typical reactions: hyperpolarization (membrane potential less than K⁺-equilibrium potential), depolarization (membrane potential higher than K⁺-potential) and absence of response at K⁺-electrochemical equilibrium. Membrane capacitance was found constant at 0.74 ± 0.05 μF cm⁻², but membrane resistance increased up to 50% independently of the sign of the electrogenic reaction. Increase of membrance capacitance and decrease of the membrane resistance was a feature of the de-energized state (ca −135 mV) and may be explained by lower viscosity of membrane lipids, which interacted with IAA. The complex of parameter, including cytoplasmic steaming taken as an indicator of energy supply, is discussed as indicating slow IAA penetration combined with a primary action of IAA on the plasmalemma receptor sites.     

Imipramine affects circadian leaf movements in Oxalis regnellii

The sensitivity of the circadian leaf movement of Oxalis regnellii Mig. to imipramine (a tricyclic dibenzazepine) was investigated. Imipramine, like Li⁺, is used as a therapeutic agent against depressive disorders in man. The therapeutic effects of the two substances might be mediated via effects on basic circadian rhythms and the cellular level. It was indeed possible to influence the circadian movement of Oxalis by imipramine; pulses (10⁻³ M , 4h) phase shifted the rhythms and caused advances. A phase response curve is presented. No period change of the movements was caused by permanent presence of imipramine (5 - 10⁻⁵ or 10⁻⁵ M ). The nature of the imipramine-induced phase shift is discussed and compared with Li⁻¹ effects on the same circadian system.     

Molybdenum-dependent degradation of nicotinic acid by Bacillus sp. DSM 2923

Abstract Growth of Bacillus sp. DSM 2923 on nicotinic acid in mineral medium was dependent on the concentration of sodium molybdate added. Addition of increasing amounts of tungstate to the medium resulted in an inhibition of growth on nicotinic acid or 6-hydroxynicotinic acid as sole source of carbon and energy. Chlorate-resistant mutants were isolated which were not able to degrade nicotinic acid and 6-hydroxynicotinic acid nor to reduce nitrate. Additionally, enzyme activities of nicotinic acid dehydrogenase and 6-hydroxynicotinic acid dehydrogenase increased with increasing concentrations of molybdate (10⁻⁸ to 10⁻⁶ M) added to the medium, and decreased with increasing amounts of tungstate (10⁻⁶ to 10⁻⁵ M) in the medium.     

Phosphate transport in potato cuttings: Effect of gibberellic acid and abscisic acid

Apical cuttings of Solanum tuberosum L. cv. Sirtema were used al different stages of development to study long-distance transport of phosphate. The effects of two hormones, gibberellic acid (GA₃) and abscisic acid (ABA), on this process were also investigated. Before tuberization, phosphate (³²P) supplied to a single leaf was transported preferentially in the young and growing parts of the plant: apical bud, young leaves and roots. After tuberization, the tuber became the principal site of phosphate accumulation. GA³ treatment (10⁻⁴ M) of the tuber as well as of the leaves led to reduced transport of ³²P into the tuber. By contrast, treatment of the tuber with ABA (10⁻⁴M) did not change the ³²P distribution within the plant, while foliar spray with ABA greatly increased the transport into the tuber. The opposite effects of the two hormones on phosphate accumulation by tubers are discussed with regard to their opposite effects on the tuberization process.     

Invertase and sucrose synthase activity, carbohydrate status and endogenous IAA levels during Citrus leaf development

Levels of activity of the sucrose catabolizing enzymes, acid invertase (EC 3.2.1.26) and sucrose synthase (EC 2.4.1.13), were measured during development of new leaves of Citrus sinensis (L.) Osbeck cv. Shamouti. Soluble acid invertase showed a peak activity of 32 nkat (g fresh weight)⁻¹ at ca 60% of full leaf expansion and rapidly declined toward and after full expansion. There was no concomitant increase in an insoluble form of the enzyme. Sucrose synthase activity, measured in the synthesis direction, declined from 33% of full leaf expansion [10 nkat (g fresh weight)⁻¹] 10, and following, full expansion. Highest sucrose synthase activity, measured in the cleavage direction, was 6 nkat (g fresh weight)⁻¹ and showed little change during development. Acid invertase has a K_m of 5 m M for sucrose, while sucrose synthase had a K_m of 118 m M for sucrose. Changes in acid invertase activity correlated with changes in the reducing sugar:sucrose ratio. These results suggest that soluble acid invertase activity is the primary enzyme responsible for sucrose catabolism in the expanding Citrus leaf. Changes in leaf expansion rate and invertase activity did not correlate positively with changes in endogenous free IAA level, as determined by enzyme linked immunoassay.     

Adventitious root formation 'in vitro' in apple rootstocks (Malus pumila) I. Factors affecting the length of the auxin-sensitive phase in M.9

The length of the auxin-sensitive phase of root initiation 'in vitro' in the apple rootstock M.9 ( Malus pumila Mill.) has been determined using the auxins indol-3yl-acettc acid (IAA) at 2.8 × 10⁻⁵ M and indol-3yl-butyric acid (IBA) at 1.5 × 10⁻⁵ M in the presence and absence of 10⁻³ M phloroglucinol (PG). PG synergised IBA-induced rooting after 4 days exposure, but contact times exceeding 8 days decreased root number. In contrast, PG consistently synergised IAA-induced rooting in the dark for contact periods up to 13 days with the highest rooting being recorded at 9 days. An irradiance of 20 W m⁻² from fluorescent lamps halved IAA-induced rooting irrespective of the presence or absence of PG. The culture of shoots at temperatures of 22,25 and 29°C during the root initiation phase (auxin present) and the root emergence phase (auxin absent) produced no difference in rooting response. In the presence of PG the use of liquid culture in place of agar-solidified culture during the auxin-sensitive phase reduced root number but not rooting percentage.     

THE RELEASE OF COPPER-COMPLEXING LIGANDS BY THE BROWN ALGA FUCUS VESICULOSUS (PHAEOPHYCEAE) IN RESPONSE TO INCREASING TOTAL COPPER LEVELS

The growth of Fucus vesiculosus L. germlings in chemically defined culture media containing a range of Cu concentrations (20–1000 nM) was monitored simultaneously with measurement of the Cu speciation in the media by competitive equilibrium-adsorptive cathodic stripping voltammetry. Fucus vesiculosus germlings were found to exude Cu-complexing ligands with conditional stability constants of the order of 1.6 × 10¹¹. Ligand concentrations increased with increasing total dissolved Cu concentrations (Cu_T) until a concentration of 500–800 neq Cu·L⁻¹ was reached. Concentrations of the ligand exceeded Cu_T in treatments containing 20 and 100 nM Cu, were similar to Cu_T in the 500-nM Cu treatment, but were less than Cu_T in the 1000-nM treatment. Therefore, [Cu²⁺] were calculated to be at concentrations of 10⁻¹¹− 10⁻¹⁰ M in the 20- and 100-nM treatments, 10⁻⁹ M in the 500-nM treatment, and 10⁻⁷ M in the 1000-nM treatment. Growth rates were lowest at Cu²⁺ concentration > 10⁻⁹. These results are discussed within the context of the potential roles for exuded copper-complexing ligands.     

Vessel differentiation along different tissue polarities

Organized vessel differentiation in an isolated system was studied on a quantitative basis. An auxin source was oriented on isolated pieces of turnip storage root ( Brassica campestris cv. Rapifera) to allow diffusion a) in the direction of, b) at right angle to, and c) opposite to the original polarity. New vessel members differentiated within 44 h, and a minimum average auxin (IAA) concentration of 3.10⁻⁶ M was required to induce initial vessel differentiation. The differentiation rates in three experimental orientations were 167, 60 and 43 cells h⁻¹ at 10⁻³ M IAA, and 1445, 1346 and 838 cells (log IAA concentration)⁻¹ after 96 h, respectively. The difference between the differentiation rates in the original polarity orientation (a) and in the orientation at right angle (b) is interpreted as reflecting reorientation itself, which requires a minimum time.     

Effects of phenolic acids on ammonia oxidation by terrestrial autotrophic nitrifying microorganisms

Abstract It has been hypothesized that vegetation in certain ecosystems inhibits nitrification in soil by producing phenolic compounds that inhibit oxidation of ammonia by nitrifying microorganisms. This hypothesis is based largely on a report that very low concentrations (10⁻⁶ M–10⁻⁸ M) of several phenolic acids (notably ferulic acid) completely inhibited NO₂⁻ production in an aqueous suspension of soil treated with (NH₄)₂SO₄ and a nutrient solution suitable for growth of Nitrosomonas and other autotrophic nitrifying microorganisms. To evaluate this hypothesis, we determined the effects of three ohenolic acids (ferulic acid, caffeic acid, and p -coumaric on nitrite production by representatives of three genera of terrestrial autotrophic nitrifying microorganisms ( Nitrosospira, Nitrosomonas , or Nitrosolubos ) grown on a defined medium containing NH₄⁺. We found that nitrite production by the Nitrososspira was not inhibited by ferulic acid, caffeic acid, or p -coumaric acid at concentrations of 10⁻⁶ or 10⁻⁵ M and was only slightly inhibited when these acids were at a concentration of 10⁻⁴ M. We also found that ferulic acid did not markedly inhibit nitrite production by the three genera of nitrifying microorganisms studied, even when its concentration was as high as 10⁻³ M. These observations invalidate the hypothesis tested because the phenolic acids studied did not significantly retard ammonia oxidation by autotrophic microorganisms even when their concentration in cultures of these microorganisms greatly exceeded their concentrations in soils.     

Effect of auxins on spermidine uptake into carrot protoplasts

The effect of an auxin, indole-3-acetic-acid (IAA), on spermidine uptake into protoplasts of carrot ( Daucus carota L. cv. Ingrid) was studied. In the presence of 1 m M Ca²⁺, IAA (10⁻⁷ to 10⁻⁴ M ) enchances [¹⁴C]-spermidine uptake into carrot protoplasts, while no stimulation occurs in the absence of Ca²⁺. The time course of the uptake with and without IAA is very rapid and reaches saturation within 1 to 2 min. Preincubation of protoplasts with IAA inhibits the spermidine uptake. La³⁺, known not to penetrate the plasmalemma, exerts the same effect as Ca²⁺, but gives lower uptake values than Ca²⁺. The application of vanadate, an ATPase inhibitor, strongly inhibits IAA-stimulated spermidine uptake, suggesting that an energy-dependent mechanism may be involved in this transport. Neither spermidine nor Ca²⁺ alone stimulate IAA uptake. The synthetic auxin 2,4-dichlorophenoxyacetic acid, yields the same results as IAA with regard to time course of spermidine uptake with and without preincubation while, unlike IAA, no significant effect was observed on the Ca²⁺ -induced increase of spermidine uptake.     

Ascorbic acid as negative effector of the peroxidase-catalyzed degradation of indole-3-acetic acid

Ascorbic acid is a strong inhibitor of indole-3-acetic oxidation catalyzed by commercial horse-radish peroxidase. In the presence of excess ascorbic acid, the indole-acetic acid oxidation catalysis is apparently blocked. The activity of peroxidase for indoleacetic acid at pH 3.7 and 33°C, in the presence of 2,4-dichlorophenol and MnCl₂ as promotors was measured by polarographic technique. The K_m was 0.27 m M and the maximum velocity was 1.02 mmol O₂ (mg protein)⁻¹ min⁻¹. Dixon plots lead to an apparent K_i of 1.25 (μ M for ascorbic acid and the inhibition was apparently competitive. Ascorbic acid, besides appearing to be a strong inhibitor of the IAA oxidase activity of peroxidase, seemed to protect IAA from total degradation. Addition of more than 5 μ M ascorbic acid produced both an exponential increase in the lag time before the onset of reaction and, at the end, an oxidation protection of 26 μ M IAA when 111 μ M IAA was present at the stawrt. The possibility of ascorbic acid-IAA auxin from endogenous oxidation in plants, is proposed.     

The Relationship of Indole-3-Acetic Acid Content and Growth of Crown-Gall Tumor Tissues of Tobacco in Culture

We have measured the content of the auxin, indole-3-acetic acid (IAA), in a cloned, crown-gall teratoma line of Nicotiana tabacum L. cv. 'Turkish' by a highly specific and sensitive radioimmunoassay. This tissue line, which does not require auxin for continuous growth in culture, exhibits two phases of growth. During the first phase, which lasts about two weeks after subculturing, growth is exponential on a fresh weight basis and the content of IAA is about 10⁻⁷–10⁻⁶ mol. kg⁻¹, but variable. During the second phase, growth rate declines gradually and the IAA content of the tissue drops dramatically; however, this drop does not result from a net loss of auxin by the tissue. The rate of growth during the exponential phase was not correlated with the IAA content of the tissue, but was strongly correlated with the IAA content of the inoculum. We found that rapidly growing leaves of Turkish tobacco have roughly the same IAA content as cultured teratoma tissues. Moreover, both tissues exhibited a similar relationship between auxin content expressed per leaf or per tissue explant and growth. These findings do not support the hypothesis that the autonomous growth of plant tumors results from an abnormally high content of auxin in the tissue.     

Effect of 2,5-norbornadiene on abscission and ethylene production in citrus leaf explants

Citrus ( Citrus sinensis L. Osbeck) leaf explants completely abscise within 48 h when exposed to saturating amounts of ethylene at 25°C. When 2,5-norbornadiene was added, 2000 μl 1⁻¹ reduced abscission of explants also exposed to 2 μl 1⁻¹ of ethylene to the level of the control, and 8000 μl 1⁻¹ reduced abscission in explants exposed to 10 μl 1⁻¹ of ethylene to the level of the control, but abscission was complete when 1 000 μl 1⁻¹ of ethylene was used in the presence of 8 000 μl 1⁻¹ of 2,5-norbornadiene. When explants were exposed to 2 μl 1⁻¹ of ethylene, 2000 μl 1⁻¹ of 2,5-norbornadiene prevented abscission if applied up to 10 h after exposure to ethylene. After 18 h, applied 2,5-norbornadiene had little effect on abscission at 48 h. A Lineweaver-Burk plot gave a 1/2 maximum value of 0.12 μl 1⁻¹ for ethylene on abscission, 2,5-Norbornadiene gave competitive kinetics with respect to ethylene with a K₁ value of approximately 120 μl 1⁻¹ of 2,5-norbornadiene. The presence of 2,5norbornadiene stimulated ethylene production, which progressively increased as the 2,5-norbornadiene concentration was increased from 250 to 8 000 μl 1⁻¹ 2,5-Norbornadiene also suppressed the induction of cellulase and polygalacturonase by ethylene. Together, 2,5-norbornadiene and 2,4-dichlorophenoxyacetic acid were more effective than either alone in reducing abscission. 2,5-Norbornadiene also was effective in preventing the reduction of indole-3-acetic acid transport induced by ethylene.     

Emission rate of amino acids and ammonia and their role in olfactory preference behaviour of juvenile Arctic charr, Salvelinus alpinus (L.)

The behaviour of juvenile Arctic charr, Salvelinus alpinus (L.) , to an abrupt concentration step of L-amino acids, L-alanine and ammonium chloride was studied by fluviarium technique. The emission rates of these substances were studied. Juvenile Arctic charr emit 8.0 × 10⁻⁴ mol total ammonia-N kg⁻¹ h⁻¹ and 3.3 × I0⁻⁵ mol amino acids kg⁻¹ h⁻¹. In behaviour tests the charr avoided 5.6x 10⁻⁶and 5.6 × 10⁻⁷ M ammonium chloride. The 17 L-amino acid mixture, ranked as observed in the analysis of emission, was avoided at 4.6 × 10⁻⁷ M, while 100 times dilution of this value gave neither avoidance nor attraction. The charr avoided L-alanine tested alone at the concentration of 4.6 × 10 ⁻⁷ M. Anosmic charr showed neither avoidance nor attraction to the mixture of 17 amino acids tested at 4.6 × 10⁻⁷ M. The results indicate that ammonia as well as emitted amino acids are not responsible for the olfactory mediated attraction to conspecific odour shown earlier in Arctic charr. On the contrary, these substances may have a negative effect by reducing the strength of attraction.     

Taste responses of the facial and glossopharyngeal nerves to amino acids in the rainbow trout

No significant differences were noted between responses of rainbow trout Oncorhynchus mykiss facial and glossopharyngeal nerves to 15 amino acids. Nine of these amino acids tested at 10⁻² M were stimulatory, whereas only two tested at 10⁻³ M were effective gustatory stimuli. For both nerve systems, ≤10⁻³ M L-proline was the most stimulatory amino acid, with an estimated threshold of 10⁻⁷ M; however, L-α-amino-β-guanidino-propionic acid (estimated threshold of 3×10⁻³ M), was the most potent compound at 10⁻² M. These results indicate that the same amino acids activate taste buds innervated by facial and glossopharyngeal nerves, respectively, and suggest that the same amino acids can be important in chemosensory feeding behaviour in the rainbow trout.