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.     

Effects of Chlorpromazine on Gravitropism in Avena Coleoptiles

Chlorpromazine (CPZ), an inhibitor of the calcium-activatedform of calmodulin, is readily taken up by the roots of intactoat seedlings but poorly translocated from the roots to thecoleoptile of these plants. However, plants repeatedly rotatedthrough solutions containing low concentrations of CPZ (10^–8–10^–5M)are infiltrated, and under these conditions, CPZ significantlyinhibits the negative gravitropic response of the coleoptilewithout retarding elongation growth. This effect is observablein ‘decapitated’ (apical 1–2 mm removed) coleoptilesections and in intact whole coleoptiles. If exogenous auxinis supplied to the decapitated sections, both their growth ratesand gravitropic responsiveness are increased and, under theseconditions, CPZ can reduce the gravitropic curvature withoutreducing the overall growth rate. These results are discussedin relation to the possible role of calmodulin-dependent calcium-ionpumps in gravitropism. chlorpromazine, gravitropism, calmodulin, calcium, oat, Avena sativa     

The Effect of Indoleacetic Acid on Cell Wall Extensibility in Avena Coleoptiles

The extensibility of cell walls in segments of living Avenacoleoptiles has been determined after treatment with IAA bythe use of a microextensometer. It has been demonstrated thatIAA has an effect both on the plastic and elastic extensibility,which is more marked if the segments are allowed to grow, exceptat temperatures nearing 0° C. The auxin-induced increasein extensibility is therefore not a direct effect on the walland metabolic factors must be involved. The results are interpreted,along cunent lines of thought, as indicating effects on thelinkage between protopectin or hemicellulose chains themselvesand between these and the microfibrillar network of cellulose.The bearing of these interpretations on the normal growth processin cell walls is briefly discussed.     

The Phototropic Responses of Avena Coleoptiles

Macleod, K., Firn, R. D. and Digby, J. 1986. The phototropicresponses of Avena coleoptiles.—J. exp. Bot. 37: 542–548. A number of studies of the elongation rate changes causing phototropismhave been made but the findings of different groups have notbeen entirely Consistent. Studies of oat coleoptile phototropismin response to first-positive and second-positive doses indicatethat no single pattern of elongation rate changes causes phototropismeven in a single species. The relative effect of phototropicstimulation on the elongation rate at the shaded or the illuminatedside of coleoptiles subject to unilateral illumination dependson physiological state of the cell-for instance its positionin the elongation zone or whether it has been given red lightrecently. Models of phototropism will have to account for sucha diversity of phototropic responses. The importance of makingfull elongation rate measurements has once again been demonstrated. Key words: Phototropism, first-positive, second-positive, phytochrome, coleoptile, elongation rate     

DMPA and Avena Coleoptiles RNA Turn-over

Addition of α-methoxy-(3, 4-dichlorophenyl) acetic acid (DMPA) at different concentrations in the culture medium of sub-apical sections of Avena coleoptile results in an increased growth rate, which correspond to the quantitative measure of the increasing of ribosomal and soluble RNA turn-over. The relative decrease in total RNA synthesis observed for the supraoptimal DMPA concentration of 1000 μg/ml appears to be due to the specifical retardation in the 4 S RNA turn-over.     

Growth and Geotropic Responses of Avena Coleoptiles to 4-Amino-3,5,6-Trichloropicolinic Acid

The elongation and geotropic responses of coleoptile sections of Avena sativa L. to various concentrations of 4-amino-3,5,6-trichloropicolinic acid (Tordon) proved to be qualitatively similar to those previously reported for 2,3,6-trichlorobenzoic acid (TCBA). Tordon stimulated growth in a range of concentrations from 1 × 10^?6 to 1 × 10^?4M but higher concentrations were inhibitory. Geotropic curvature was extensively depressed by 1 × 10^?5 and 1 × 10^?4M Tordon, concentrations which accelerated elongation. A similar differential effect has been reported for TCBA and other auxins. Several other picolinic acids and related compounds were tested, but only very slight responses were noted.     

Some Responses of Avena Coleoptiles to Ethylene

Ethylene pretreatment of intact Avena seedlings or of excisedcoleoptile sections results in an increased response of thecells to auxin. It is suggested that ethylene brings about theacceleration of hydrolytic reactions controlling the physicalproperties of cell walls and hence increases their capacityfor growth. Coleoptile elongation of intact seedlings is inhibitedby ethylene; this inhibition is concurrent with a lateral expansionof the entire coleoptile. It is suggested that under a givenset of conditions coleoptile cells are capable of attaininga finite volume and that the preferential lateral expansioninduced by ethylene is accomplished at the expense of longitudinalextension. Experiments with intact and deseeded plants indicatethat lateral expansion depends on the supply of some factorfrom the endosperm.     

Phytochrome-controlled Hydrogen Ion Excretion by Avena Coleoptiles

A red light-induced, far red reversible stimulation of proton efflux from apical segments of etiolated Avena sativa L. cv. Victory coleoptiles was observed. The acidification responses to red light and also to auxin were not the consequence of respired CO(2). The response to red light was strongly inhibited by cycloheximide and carbonyl cyanide, m-chlorophenyl hydrazone, but mannitol had a stimulatory effect. Red light and auxin applied together yielded a greater than additive response, in comparison to the effects of the two stimuli applied separately.     

pH-Dependence of Extension Growth in Avena Coleoptiles and Its Implications for the Mechanism of Auxin Action

The pH-dependence of acid-induced growth in excised segments of Avena sativa coleoptiles has been reinvestigated in the pH range 3 to 7. In contrast to previous reports (e.g. DL Rayle [1973] Planta 114: 63-73), only acidic buffers with a pH below 5.0 induce an extension response. A pH of 3.5 to 4.0 is required to mimic auxin-mediated growth. Very similar pH-response curves are obtained with both intact (abraded) and peeled coleoptiles. These results agree with the recent finding of a similarly low sensitivity to protons in maize coleoptiles. It is shown that the apparently much higher sensitivity to protons previously reported for peeled Avena coleoptiles is due to incubating the tissue in buffer of pH 6.8 between peeling and measuring the effect of acidic buffers. Neutral pH reversibly inhibits the spontaneous extension burst originating on release from tissue tension after removing the epidermis. Reversal of this inhibition can be achieved by buffers of pH 5.0 to 6.0 (or distilled water), thereby simulating an acid-induced growth response in this pH range. It is concluded that true acid-induced wall-loosening generally does not take place above pH 5.0 and that a pH considerably below 4.0 is required in order to stimulate growth to an extent comparable to that obtained in response to auxin. The “acid-growth theory,” which requires an acid-mediated loosening of the cell wall in the pH range 5 to 6, this pH being established by auxin-induced proton excretion, can therefore also not be substantiated in Avena.     

Calcium Requirement for Indoleacetic Acid-induced Acidification by Avena Coleoptiles

The ionic specificity of IAA-induced acidification by Avena coleoptiles was studied, using zwitterionic, presumably impermeant buffers. The acidification was almost totally dependent on divalent cations with an order of effectiveness of Ca(2+) >/= Sr(2+) > Mn(2+), Mg(2+); whereas other polyvalent cations tested were ineffective. The Ca(2+) response was IAA-dependent. The CaCl(2) concentration was optimal at 0.3 to 1 mm and inhibitory at higher concentrations. Sr(2+) inhibited Ca(2+)-dependent acidification and monovalent cations such as K(+) did not induce additional acidification in the presence of optimal CaCl(2). These data are consistent with a mechanism for IAA-induced acidification involving a Ca(2+) -H(+) exchange.     

Inhibition of Low pH-induced Elongation in Avena Coleoptiles by Abscisic Acid

An angular position-sensing transducer was used to make continuous measurements of acid-induced elongation of Avena sativa coleoptile segments. Elongation rates at pH 4.5 (5 mm succinate buffer) were about 5-fold greater than those at pH 6.0. Buffered 0.1 mm abscisic acid produced a partial decrease of the growth rate. Pretreatments with abscisic acid buffered at pH 6.0 usually caused a further reduction of the elongation response when the coleoptile segments were subsequently placed in buffer at pH 4.5 containing abscisic acid. Abscisic acid did not completely prevent the pH effect in any of these experiments, and the brief latent period of the pH response was not affected by abscisic acid treatments. At pH 4.5, where the inhibitory effect of ABA was maximum, low pH-induced elongation was also inhibited by KCN and HgCl₂. These results suggest that pH-(4.5) induced elongation in this system may be dependent on some metabolic processes and that abscisic acid-induced inhibition of this elongation may involve an interaction with these processes.