Effect of temperature on the dose–response curves for auxin-induced elongation growth in maize coleoptile segments

The dose–response curves for IAA-induced growth in maize coleoptile segments were studied as a function of time and temperature. In addition, the kinetics of growth rate responses at some auxin concentrations and temperatures was also compared. It was found that the dose–response curves for IAA-induced elongation growth were, independently of time and temperature, bell-shaped with an optimal concentration at 10⁻⁵ M IAA. The kinetics of IAA-induced growth rate responses depended on IAA concentration and temperature, and could be separated into two phases (biphasic reaction). The first phase (very rapid) was followed by a long lasting one (second phase), which began about 30 min after auxin addition. For coleoptile segments incubated at 30°C, the amplitudes of the first and second phase were significantly higher, when compared with 25°C, at all IAA concentrations studied. However, when coleoptile segments were incubated at 20°C, the elongation growth of coleoptile segments treated with suboptimal IAA concentrations was diminished, mainly as a result of both phases reduction. In conclusion, we propose that the shape of the dose–response curves for IAA-induced growth in maize coleoptile segments is connected with biphasic kinetic of growth rate response.     

Stimulatory effects of potassium iodide on auxin action in the coleoptile segments of maize (Zea mays)

Potassium iodide (KI) was found to stimulate IAA-induced elongation of coleoptile segments in maize (Zea mays L.). The promoting effects of KI on coleoptile elongation, which were optimal at 1 mM in the presence of IAA, did not occur as a result of better conservation of IAA in the incubation medium. In addition, KI did not affect fusicoccin- or epibrassinolide-induced elongation. Additionally, sodium iodide (NaI) induced similar stimulatory effects on IAA-induced elongation, however, potassium chloride (KCl) showed no effect, suggesting that iodide is the active component. KI also enhanced IAA-induced ethylene biosynthesis in maize coleoptile segments. Taken together, these results suggest the involvement of KI-sensitive step(s) in auxin action before effectors of the signal transduction pathway split to elongation growth and ethylene biosynthesis. In-yong Hwang and Soo Chul Chang contributed equally to this work.     

Effects of UV-C radiation on extension growth, H+-extrusion and transmembrane electric potential in maize coleoptile segments

The effect of 253.7 nm ultraviolet radiation on elongation growth, medium acidification and changes in electric potential difference between vacuole and external medium in cells of maize ( Zea mays L.) coleoptile segments was investigated. It was found that irradiation with 390, 1170, 3900 and 5 850 J m⁻² UV-C (ultraviolet radiation 253.7 nm) inhibited elongation growth, whereas at 195 J m⁻² stimulation of growth was observed. The administration of IAA (10⁻⁵ M ) to the incubation medium of coleoptile segments partially abolished the inhibitory effect of UV-C. The pH of the incubation medium, measured simultaneously with growth, showed that the exposure of the segments to UV-C caused inhibition of H⁺-extrusion (or stimulation of H⁺ uptake). The presence of IAA (10⁻⁵ M ) in the incubation medium promoted (except after 5850 J m⁻² irradiation) H⁺-extrusion to a level comparable with that produced by IAA in non-irradiated segments. In UV-C irradiated segments the potential difference underwent significant alterations. Irradiation of coleoptile segments with 390 J m⁻² caused a transient depolarization, which was fully reversible within 30 min, while at higher doses depolarization was irreversible. The hyperpolarization of the membrane potential (MP) in cells of maize coleoptile induced by IAA was completely nullified by subsequent irradiation with UV-C. It is suggested that UV-C inhibited IAA-induced growth by a mechanism independent of cell wall acidification.     

Effect of cadmium on growth, proton extrusion and membrane potential in maize coleoptile segments

Cd accumulation, its effects on elongation growth of maize coleoptile segments, pH changes of their incubation medium and the membrane potential of parenchymal cells were studied. The Cd content increased significantly with exposure to increasing cadmium concentrations. Coleoptile segments accumulated the metal more efficiently in the range 10–100 μM Cd, than in the range 100–1000 μM Cd. Cd at concentrations higher than 1.0 μM produced a significant inhibition of both growth and proton extrusion. 100 μM Cd caused depolarization of the plasma membrane (PM) potential in parenchymal cells. The simultaneous treatment of maize coleoptile segments by indole-3-acetic acid (IAA) and Cd, counteracted the toxic effect of Cd on growth. Moreover, our data also showed that 100 μM Cd suppressed the characteristic IAA-induced hyperpolarization of the membrane potential, causing membrane depolarization. These results indicate that the toxic effect of Cd on growth of maize coleoptile segments might be, at least in part, caused via reduced PM H⁺-ATPase activity.     

Cellular responses to naphthoquinones: juglone as a case study

The effects of juglone (JG) on the endogenous growth, growth in the presence of either indoleacetic acid (IAA) or fusicoccin (FC) and on proton extrusion were studied in maize coleoptile segments. In addition, membrane potential changes were also determined at chosen JG concentrations. It was found that JG, when added to the incubation medium, inhibited endogenous growth as well as growth in the presence of either IAA or FC. Simultaneous measurements of growth and external pH indicated that inhibition of either IAA-induced growth or proton extrusion by JG was a linear function of JG concentration. Addition of JG to the control medium caused depolarization of the membrane potential (E_m), value of which was dependent on JG concentration and time after its administration. Hyperpolarization of E_m induced by IAA was suppressed in the presence of JG. It was also found that for coleoptile segments initially preincubated with JG, although subsequently removed, addition of IAA was not effective in the stimulation of growth and medium acidification. Taken together, these results suggest that the mechanism by which JG inhibits the IAA-induced growth of maize coleoptile segments involves inhibition of PM H⁺-ATPase activity.     

Role of chloride ions in the promotion of auxin-induced growth of maize coleoptile segments

Background and Aims

The mechanism of auxin action on ion transport in growing cells has not been determined in detail. In particular, little is known about the role of chloride in the auxin-induced growth of coleoptile cells. Moreover, the data that do exist in the literature are controversial. This study describes experiments that were carried out with maize (Zea mays) coleoptile segments, this being a classical model system for studies of plant cell elongation growth.

Methods

Growth kinetics or growth and pH changes were recorded in maize coleoptiles using two independent measuring systems. The growth rate of the segments was measured simultaneously with medium pH changes. Membrane potential changes in parenchymal cells of the segments were also determined for chosen variants. The question of whether anion transport is involved in auxin-induced growth of maize coleoptile segments was primarily studied using anion channel blockers [anthracene-9-carboxylic acid (A-9-C) and 4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid (DIDS)]. In addition, experiments in which KCl was replaced by KNO₃ were also performed.

Key Results

Both anion channel blockers, added at 0·1 mm, diminished indole-3-acetic acid (IAA)-induced elongation growth by ∼30 %. Medium pH changes measured simultaneously with growth indicated that while DIDS stopped IAA-induced proton extrusion, A-9-C diminished it by only 50 %. Addition of A-9-C to medium containing 1 mm KCl did not affect the characteristic kinetics of IAA-induced membrane potential changes, while in the presence of 10 mm KCl the channel blocker stopped IAA-induced membrane hyperpolarization. Replacement of KCl with KNO₃ significantly decreased IAA-induced growth and inhibited proton extrusion. In contrast to the KCl concentration, the concentration of KNO₃ did not affect the growth-stimulatory effect of IAA. For comparison, the effects of the cation channel blocker tetraethylammonium chloride (TEA-Cl) on IAA-induced growth and proton extrusion were also determined. TEA-Cl, added 1 h before IAA, caused reduction of growth by 49·9 % and inhibition of proton extrusion.

Conclusions

These results suggest that Cl^– plays a role in the IAA-induced growth of maize coleoptile segments. A possible mechanism for Cl^– uptake during IAA-induced growth is proposed in which uptake of K⁺ and Cl^– ions in concert with IAA-induced plasma membrane H⁺-ATPase activity changes the membrane potential to a value needed for turgor adjustment during the growth of maize coleoptile cells.     

The dose-response curves for IAA induced elongation growth and acidification of the incubation medium of Zea mays coleoptile segments

The initial dose-response curves for auxin-induced elongation growth of Zea mays L. coleoptile segments and simultaneously measured changes of pH of the incubation medium were studied. It was found that these curves are bell-shaped on all occasions and that at all IAA concentrations studied acidification of the incubation medium took place. The optimum response for IAA-induced elongation growth and acidification of the incubation medium was 10⁻⁵ and 10⁻⁴ M IAA, respectively. The regression curves and correlation coefficients between magnitude of the growth response and acidification of the incubation medium indicated a close relationship between these sets of data over a wide range of IAA concentrations.     

Effect of cycloheximide on IAA-induced proton excretion and IAA-induced growth in abraded Avena coleoptiles

IAA-induced proton excretion in peeled or abraded oat ( Avena saliva L. cv. Victory) coleoptiles is closely associated with IAA-induced growth. It was attempted to separate these two processes by using cycloheximide to inhibit them differentially. Growth of abraded coleoptile segments was measured by a shadow graphic method, and their IAA-induced acidification of the external solution was monitored with a pH meter. IAA stimulated proton excretion in abraded Avena coleoptile segments after a 13 min lag. IAA-induced proton excretion was inhibited within 5 min by cycloheximide at concentrations of 1.8 × 10⁻⁶, 3.6 × 10 or 3.6 × 10⁻⁵ M. Cycloheximide at these concentrations, added within 4 min of IAA, prevented IAA-induced acidification of the medium for at least 60 min. However, it did not prevent IAA-induced growth during this time. It is concluded that some of the initial IAA-induced growth seen in Avena coleoptiles is independent of detectable IAA-induced proton excretion.     

Effect of temperature on growth,proton extrusion and membrane potential in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) coleoptile segments

The effects of temperature (5–45°C) on endogenous growth, growth in the presence of either indoleacetic acid (IAA) or fusicoccin (FC), and proton extrusion in maize coleoptile segments were studied. In addition, membrane potential changes at some temperatures were also determined. It was found that in this model system endogenous growth exhibits a clear maximum at 30°C, whereas growth in the presence of IAA and FC shows the maximum value in the range 30–35°C and 35–40°C, respectively. Simultaneous measurements of growth and external medium pH indicated that FC at stressful temperatures was not only much more active in the stimulation of growth, but was also more effective in acidifying the external medium than IAA. Also the addition of either IAA or FC to the bathing medium at 30 and 40°C did not change the kinetic characteristic of membrane potential changes observed for both substances at 25°C. However, the increased temperature significantly decreased IAA and FC-induced membrane hyperpolarization. IAA in the incubation medium, at 10°C, brought about additional membrane depolarization (apart from the one induced by low temperature). In contrast to IAA, FC at 10°C caused gradual repolarization of membrane potential, which correlated with both FC-induced growth and FC-induced proton extrusion. A plausible interpretation for temperature-induced changes in growth of maize coleoptile segments is that, at least in part, these changes were mediated via a PM H⁺-ATPase activity.     

Effect of thiosulphinates contained in garlic extract on growth, proton fluxes and membrane potential in maize (Zea mays L.) coleoptile segments

The effect of thiosulphinates contained in garlic extract (GE) on endogenous growth, growth in the presence of either indoleacetic acid (IAA) or fusicoccin (FC), and proton extrusion in maize coleoptile segments were studied. In addition, membrane potential changes at some GE dilutions and the protective effect of dithiothreitol (DTT) against GE toxicity were also determined. It was found that GE at almost all dilutions studied, when added to the incubation medium inhibited endogenous growth as well as growth in the presence of either IAA or FC. Simultaneous measurements of growth and external pH indicated that the administration of GE resulted in a complex change in the pH of the external medium; after an initial transient acidification, pH increased and reached the maximal value followed by a gradual decrease of medium pH. When IAA or FC was added after preincubation of the segments in the presence of GE the changes in medium pH were not significantly different from these obtained with GE only. If the coleoptile segments were first preincubated with GE and subsequently GE was removed, the addition of IAA induced strong growth and medium acidification. Dithiothreitol added together with GE neutralized the toxic effect of GE on growth of coleoptile segments incubated in the presence of IAA. The addition of GE to the control medium caused a depolarization of the membrane potential, the value of witch depended on GE dilution. These results indicate that the toxic effect of GE on growth of plant cells might be caused by disruption of the catalytic function of the plasma membrane H⁺-ATPase on formation of the disulfide bonds.     

Fusicoccin counteracts inhibitory effects of high temperature on auxin-induced growth and proton extrusion in maize coleoptile segments

Plant growth and development are tightly regulated by both plant growth substances and environmental factors such as temperature. Taking into account the above, it was reasonable to point out that indole-3-acetic acid (IAA), the most abundant type of auxin in plants, could be involved in temperature- dependent growth of plant cells. We have recently shown that growth of maize coleoptile segments in the presence of auxin (IAA) and fusicoccin (FC) shows the maximum value in the range 30–35°C and 35–40°C, respectively. Furthermore, simultaneous measurements of growth and external medium pH indicated that FC at stressful temperatures was not only much more active in the stimulation of growth, but was also more effective in acidifying the external medium than IAA. The aim of this addendum is to determine interrelations between the action of IAA and FC (applied together with IAA) on growth and medium pH of maize coleoptile segments incubated at high temperature (40°C), which was optimal for FC but not for IAA.Key words: auxin, fusicoccin, coleoptile segments, elongation growth, medium pHA well studied aspect of auxin action especially in maize coleoptile, is its effect on cell elongation, proton extrusion and membrane potential.^1–7 It is now generally agreed that indole-3-acetic acid (IAA), as the principal regulator of plant elongation growth, causes (i) acceleration of elongation growth as compared to endogenous growth, (ii) enhancement of proton extrusion as compared to auxin—free medium, and (iii) transient depolarization followed by a slow hyperpolarization of membrane potential. According to the “acid growth theory” of elongation growth,^8–11 auxin induced cell wall acidification provides favorable conditions for cell wall loosening, a requirement for cell elongation. At least in maize coleoptile segments, auxin induced cell wall acidification is mediated by increased activity and/or amount of the PM H⁺-ATPase.^11,12 In the case of fusicoccin, which mimics the effect of auxin in many respects,¹³ it was shown that FC-binding site arises from interaction of the 14-3-3 protein dimmer with the C-terminal autoinhibitory domain of the H⁺-ATPase and that FC stabilizes this complex.^14–18 It should be pointed out that in spite of abundant literature on the mechanism through which IAA or FC control growth of grass coleoptiles, little is know how these substances work at extreme temperatures. Over the past decade, the involvement of 14-3-3 proteins in plant stress responses has often been suggested.¹⁹ For example, work by Chelysheva et al.,²⁰ and Babakov et al.,²¹ demonstrated that under low temperature and high osmolarity conditions, 14-3-3 proteins interact with the C-terminal autoinhibitory domain of the PM H⁺-ATPase activating the proton pump that play a key role in stress responses in higher plants. We have recently shown²² that FC at 40°C induced maximal growth whereas growth observed at the same temperature in the presence of IAA was reduced by 33% compared to the maximal value at 30°C. It was also found²² that at 40°C the kinetics of the pH change differed significantly for both growth substances; the segments treated with IAA at 40°C were virtually not able to acidify the external medium, whereas FC at this temperature caused practically maximal acidification. In this addendum we have shown that application of FC together with IAA conteracted the inhibitory effect of high temperature (40°C) on IAA-induced growth and proton extrusion in maize coleoptile segments (Fig. 1). For example, the total IAA-induced elongation growth of coleoptile segments at 40°C was 1438.1 ± 134.5 µm cm⁻¹ (mean ± SE, n = 11) while elongation of 2747.4 ± 269.7 µm cm⁻¹ (mean ± SE, n = 11) was observed in IAA applied together with FC (Fig. 1A). The data in Figure 1B indicate that coleoptile segments incubated at 40°C (over 2 h), without growth substances (control) characteristically changed the pH of the medium: usually within the first 30–45 min an increase of pH (by ca. 0.5 pH unit) was observed, followed by a slow decrease of pH. When IAA or FC was added (after 2 h of segment''s incubation in control medium), an additional decrease of pH was observed. As can be seen in Figure 1B, FC added at 40°C was much more effective in acidification of the medium, as compared to IAA. For FC, 5h after its addition, the pH of the incubation medium dropped to pH 4.2, whereas for IAA the pH was only 5.4. However, addition of IAA together with FC at 40°C dropped medium pH approximately to the same value as was observed in the presence of FC only.Open in a separate windowFigure 1Effect of high temperature (40°C) on growth (A) and medium pH (B) of maize coleoptile segments incubated in the presence of IAA (10 µM) and FC (1 µM). The growth of a stack of 21 segments, expressed as elongation (µm cm⁻¹), was measured simultaneously with medium pH at 40°C. After preincubation (over 2 h) of the coleoptile segments in control medium, IAA and FC was added (arrow). Values are means of 11 independent experiments. Bars indicate ± SE. In the case of medium pH SE did not exceed 8%.In conclusion, the results presented in this addendum provide further evidence that FC on the receptor level is much more effective than IAA.     

Auxin-induced elongation growth and expressions of cell wall-bound exo- and endo-beta-glucanases in barley coleoptiles

When auxin stimulates rapid cell elongation growth of cereal coleoptiles, it causes a degradation of 1,3:1,4-beta-glucan in hemicellulosic polysaccharides. We examined gene expressions of endo-1,3:1,4-beta-glucanase (EI) and exo-beta-glucanase (ExoII), of which optimum pH are about 5, and molecular distribution of hemicellulosic polysaccharides in barley (Hordeum vulgare L.) coleoptile segments treated with or without IAA. IAA (10(-5) M) stimulated the gene expression of EI, while it did not affect that of ExoII. IAA induced gene expression of EI after 4 h and increased wall-bound glucanase activity after 8 h. The molecular weight distribution of hemicellulosic polysaccharides from coleoptile cell walls was shifted to lower molecular weight region by 2 h of IAA treatment. Fusicoccin (10(-6) M) mimicked IAA-induced elongation growth and the decrease in molecular weight of hemicellulosic 1,3:1,4-beta-glucan of coleoptiles in the first 4 h, but it did not promote elongation growth thereafter. These facts suggest that acidification of barley cell walls by IAA action enhances pre-existing cell wall-bound glucanase activity in the early first phase of IAA-induced growth and the late second phase involves the gene expression of EI by IAA.     

A comparison of the effects of 1,4-naphthoquinone and 2-hydroxy-1,4-naphthoquinone (lawsone) on indole-3-acetic acid (IAA)-induced growth of maize coleoptile cells

The effects of 1,4-naphthoquinone (NQ) and 2-hydroxy-1,4-naphthoquinone (NQ-2-OH) on indole-3-acetic acid (IAA)-induced growth, medium pH changes and membrane potential (E_m) in maize (Zea mays L.) coleoptile cells were determined. In addition, the redox cycling properties of both naphthoquinones were also compared. The dose-response curves constructed for the effects of NQ and NQ-2-OH on endogenous and IAA-induced growth differ in shape. It was found that NQ was by 10–50% more effective in inhibiting IAA-induced growth in maize coleoptile segments than NQ-2-OH. Simultaneous measurements of growth and external medium pH indicated that NQ and NQ-2-OH reduced or eliminated proton extrusion at all of the concentrations used, excluding NQ at 1 µM. It was found that both naphthoquinones at concentrations higher than 10 µM caused the depolarisation of the membrane potential (E_m). Additionally, compared to the controls, NQ- and NQ-2-OH-exposure of coleoptile segments, at concentrations higher than 10 µM, caused an elevation of the hydrogen peroxide (H₂O₂) production and plasma membrane redox activity. The highest catalase activity was observed at 10 µM NQ and it was ca. 18-fold greater (at 4 h) than in the control medium. Moreover, it was also found that NQ and NQ-2-OH, at all concentrations studied, increased the malondialdehyde content of coleoptile segments at 4 h of the experiment. The data presented here are discussed taking into account the “acid growth hypothesis” of auxin action and the mechanisms by which naphthoquinones interact with biological systems.     

Galactose prevents proton excretion but not IAA-induced growth in segments of azuki bean epicotyls

We investigated the effect of galactose on IAA-induced elongation and proton excretion in azuki bean (Vigna angularis Ohwi et Ohashi) segments in order to confirm whether or not protons were involved in auxin-induced growth. Galactose inhibited the IAA-induced decrease in the solution pH but had no inhibitory effect on IAA-induced growth in segments of azuki bean epicotyls. On the other hand, galactose inhibited both IAA-induced growth and proton excretion in oat (Avena sativa L.) coleoptile segments. From these results it is unlikely that IAA-induced growth is mediated by proton excretion at least in azuki bean epicotyls.Abbreviations IAA indole-3-acetic acid - FC fusicoccin     

A comparison of the effects of IAA and 4-Cl-IAA on growth, proton secretion and membrane potential in maize coleoptile segments

The physiological activity of exogenous 4-Cl-IAA, as compared to IAA, was examined in maize coleoptile segments. It was found that in this model system 4-Cl-IAA is much more active in the stimulation of elongation than IAA. Simultaneous measurements of growth and external pH indicated that administration of either IAA or 4-Cl-IAA resulted in medium acidification. The kinetics of the pH changes, however, were faster after the addition of 4-Cl-IAA. In contrast to IAA, the coleoptile segments treated with chlorinated auxin were not able to increase medium pH after its initial drop. The re-addition of IAA after 5 h further enhanced growth over the next 2 h by 31%. By contrast, the re-addition of 4-Cl-IAA at the same time protocol as IAA did not cause an additional effect. The administration of 10 microM IAA induced in maize coleoptile cells a transient depolarization followed by a slow hyperpolarization of their membrane potential. In contrast to IAA, 4-Cl-IAA at 1 microM caused an immediate hyperpolarization of the membrane potential which, on average, was 2-fold greater than for IAA. The results reported here provide further evidence that 4-Cl-IAA is much more active, as compared to IAA, in stimulating the growth of maize coleoptile segments. Although it has not been directly demonstrated here, a plausible interpretation for the high 4-Cl-IAA activity is that, at least in part, it might be caused via a reduced metabolism of 4-Cl-IAA. Furthermore, for the first time, the data show that membrane potential responds to 4-Cl-IAA in a qualitatively different fashion than to IAA. These findings may, in turn, suggest a specific signal transduction pathway to 4-Cl-IAA in maize coleoptile cells.     

The effects of electric field on the growth of intact seedlings and coleoptile segments ofZea mays L.

The experiments were carried out with 96-h-old intact maize seedlings and 10 mm long coleoptile segments cut 4 mm below the tip. The electric fields were applied longitudinally along the seedlings. The electric field (15 V) caused inhibition of the elongation growth of intact seedlings which was dependent on both the polarity and the duration of the applied voltage. The growth inhibition was greater when the tip of the shoot was positive relative to the roots. The electric field also caused inhibition of indole-3-acetic acid (IAA) and fusicoccin (FC) induced growth of maize coleoptile segments excised from electrically treated seedlings. IAA-induced growth of coleoptile segments was greater when the tip of the shoot was negative to the roots (not in the case of FC-treated segments and intact seedlings). It was suggested that apart from the changes induced by electric field in transport system of auxin the electric field affected also the activity of plasmalemma proton pump.     

UDP-Glucose level as a limiting factor for IAA-induced cell elongation in Avena coleoptile segments

The effects of galactose on IAA-induced elongation and endogenous level of UDP-glucose (UDPG) in oat ( Avena sativa L. cv. Victory) coleoptile segments were examined under various growth conditions to see if there was a correlation between the level of UDPG and auxin-induced growth. The following results were obtained:

• (1)

  Galactose (10 m M ) inhibited the auxin-induced cell elongation of oat coleoptile segments after a lag of ca 2 h. Determinations of cell wall polysaccharides and UDP-sugars indicated that galactose, when inhibiting the cell wall polysaccharide synthesis, decreased the level of UDPG but caused an increase in the levels of Gal-1-P and UDP-Gal.
• (2)

  When coleoptile segments treated with IAA and galactose were transferred to galactose-free IAA-solution, the segment elongation was restored and the amounts of cell wall polysaccharides increased. During this period, the amount of UDPG increased and the levels of Gal-1-P and UDP-Gal slightly decreased or leveled off. The UDP-pentoses changed similarly as UDPG did.
• (3)

  Addition of sucrose (30 m M ) enhanced IAA-induced cell elongation and removed growth inhibition by 1 m M galactose. Sucrose increased the amounts of the cell wall polysaccharides and the level of UDPG in the presence or absence of IAA and also counteracted the decrease in UDPG caused by galactose.

These results indicate that the level of UDPG is an important limiting factor for cell wall biosynthesis and, thus, for auxin-induced elongation.     

Determination of Auxin-Dependent pH Changes in Coleoptile Cell Walls by a Null-Point Method

The present debate on the validity of the "acid-growth theory" of auxin (indole-3-acetic acid, IAA) action concentrates on the question of whether IAA-induced proton excretion into the cell wall is quantitatively sufficient to provide the shift in pH that is required to explain IAA-induced growth (see D.L. Rayle, R.E. Cleland [1992] Plant Physiol 99:1271-1274 for a recent apologetic review of the acid-growth theory). In the present paper a null-point method has been employed for determining the growth-effective cell-wall pH in the presence and absence of IAA after 60 min of treatment. Elongation of abraded maize (Zea mays L.) and oat (Avena sativa L.) coleoptile segments was measured with the high resolution of a displacement transducer. The abrasion method employed for rendering the outer epidermal cell wall permeable for buffer ions was checked with a dye-uptake method. Evidence is provided demonstrating that externally applied solutes rapidly and homogeneously penetrate into the epidermal wall, whereas penetration into the inner tissue walls is strongly retarded. "Titration" curves of IAA-induced and basal elongation were determined by measuring the promoting/inhibiting effect of medium pH under iso-osmotic conditions in the range of pH 4.5 to 6.0. In maize, the null point (no pH-dependent change in elongation rate after 5-10 min of treatment with 10 mmol L-1 citrate buffer) was pH 5.00 after 60 min of IAA-induced growth, and the null-point pH determined similarly in IAA-depleted tissue (10 times smaller elongation rate) was 5.25. Corresponding titration curves with Avena segments led to slightly lower null-point pH values both in the presence and absence of IAA-induced growth. After induction of acid-mediated extension by 1 [mu]mol L-1 fusicoccin (FC) in maize, the null-point pH shifted to 3.9. At 0.5 [mu]mol L-1, FC induced the same elongation rate as IAA but a 9-fold larger rate of proton excretion. At 0.033 [mu]mol L-1, FC induced the same rate of proton excretion as IAA but had no appreciable effect on elongation. The implications of these results against the background of recent attempts to revitalize the acid-growth theory of IAA action are discussed.     

Jasmonic Acid Inhibits the IAA-Induced Elongation of Oat Coleoptile Segments: a Possible Mechanism Involving the Metabolism of Cell Wall Polysaccharides

The effects of jasmonic acid (JA) on the IAA-induced elongationof segments of etiolated oat (Avena sativa L. cv. Victory) coleoptileswere studied. Exogenously applied JA substantially inhibitedIAA-induced elongation of oat coleoptile segments. The inhibitionof the growth of oat coleoptile segments due to JA appeared2 h after the application of JA with IAA. JA did not affectthe consumption of oxygen by the segments, the osmolarity ofthe cell sap or the IAA-induced loosening of cell walls, whichwas recognized as a decrease in the minimum stress-relaxationtime (T₀). JA was extremely effective in preventing increasesin the amount of the cell wall polysaccharides in both the non-cellulosicfraction and the cellulosic fraction during coleoptile growthin the presence and in the absence of IAA. Inhibition of thegrowth of oat coleoptile segments induced by JA was partiallyreversed by the simultaneous addition of sucrose to the testsolution. From these results, it appears that JA inhibits IAA-inducedelongation of oat coleoptile segments by interfering with someaspects of sugar metabolism that are related to the degradationand/or the synthesis of cell wall polysaccharides. (Received March 15, 1994; Accepted August 2, 1994)