Waldemar Karcz

Lead distribution in corn seedlings (Zea mays L.) and its effect on growth and the concentrations of potassium and calcium

It was observed that dry weight yield is not a sensitive parameter withwhich to assess lead toxicity to plants. Elongation growth of corn seedlingroots was more sensitive to lead than shoot growth and was inhibited by allconcentrations tested (10−5, 10−4, and 10−3 M).It was positively correlated with potassium concentration and negativelycorrelated with lead concentration in the roots. Negative correlation also wasobserved between lead concentration and potassium concentration in roots. It ispostulated that inhibition of corn root growth is connected with potassiumleakage from root cells. The toxic action of lead on corn seedling mesocotylandcoleoptile growth was not correlated with potassium concentration in planttissue and correlation between growth and lead concentration was low. Inseedlings treated with 10−4 and 10−3 M lead the growthof mesocotyl and coleoptile was affected similarly, although the concentrationof lead was threefold higher in mesocotyl tissue than in coleoptile tissue. It isproposed that depression of corn seedlings shoot growth is not an effect ofpotassium leakage or lead accumulation but of an unknown signal induced inroots, as a response to exposure to lead, which is transmitted to shoots. Thepositive correlation between lead and calcium concentrations found in seedlingroots might be connected with high constitutional tolerance of corn to lead.Since the first 8 mm of an apical root accounts for 50% of thelead accumulated by the whole root, it is postulated that rhizofiltration oflead contaminated waters should be more efficient when plant species withhighly branched root systems are used.

Effect of temperature on growth, proton extrusion and membrane potential in maize (Zea mays 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.

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 (Em), value of which was dependent on JG concentration and time after its administration. Hyperpolarization of Em 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.

Effect of cadmium and lead on the membrane potential and photoelectric reaction of Nitellopsis obtusa cells.

The effects of Cd and Pb on membrane potential (E(m)) and photoelectric reaction of Nitellopsis obtusa cells were investigated. It was found that Cd and Pb at 1.0 mM caused a depolarization of the E(m), whereas both metals at lower concentrations changed the E(m) in a different way. Pb at 0.1 mM and 0.01 mM hyperpolarized the E(m), whereas Cd at the same concentrations depolarized and did not change the E(m), respectively. In the presence of 0.01 mM Pb, the light-induced hyperpolarization of the E(m) was by 18% higher as compared to the control, whereas at 1.0 mM Pb it was by 40% lower. Pb at 0.1 mM and Cd at 0.01 mM or 5 × 0.01 mM did not change the light-induced membrane hyperpolarization. However, in the presence of Cd at 0.1 mM and 1.0 mM this hyperpolarization was 2-fold lower or was completely abolished, respectively. These results suggest that at high Cd and Pb concentrations both depolarization of the E(m) and decrease of light-induced membrane hyperpolarization in Nitellopsis obtusa cells are probably due to inhibition of the plasma membrane H(+)-ATPase activity, whereas both metals at lower concentrations differ in mechanism of membrane potential changes.

Effect of IAA and 4-Cl-IAA on growth rate in maize coleoptile segments

The dose-response curves for IAA and 4-Cl-IAA-induced growth of Zea mays L. coleoptile segments were studied as a function of time. Moreover, some characteristic growth parameters for both auxins were compared. The dose-response curve of growth rate measured after IAA or 4-Cl-IAA application was bell-shaped in all experiments. The optimum concentration was 10−6 M for 4-Cl-IAA and was found not to depend on the time of the growth measurement. However, in the case of IAA the optimum shifted from 10−6 M at the time of maximal growth rate to 10−5 M or even 10−4 M, when growth measured 3–4 hours after auxin application was analysed. The relative activity of 4-Cl-IAA-induced growth rate (as compared to IAA) increased significantly with increasing time from addition of this auxin to the medium. For both auxins the time needed to reach the maximal growth rate was clearly related to their concentrations. These data provided further evidence that 4-Cl-IAA is much more active auxin than IAA and can also suggest that IAA is more rapidly metabolized in comparison to 4-Cl-IAA.

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.

Interactive effects of temperature and heavy metals (Cd, Pb) on the elongation growth in maize coleoptiles.

The effect of Cd and Pb on endogenous and IAA-induced elongation growth and medium pH of maize coleoptile segments incubated at 20, 25 and 30 °C was studied. It was found that the elongation of coleoptile segments and proton extrusion increased with the temperature and reached its maximum at 30 °C. For Cd, the maximal inhibition of endogenous and IAA-induced growth as well as medium acidification of coleoptile segments was observed at 25 °C. Meanwhile, Pb, irrespective of the temperature, diminished the growth of the segments by ca. 20%, increasing the acidification of the incubation medium. It was also found that in contrast to Cd, Pb accumulation in maize coleoptile segments did not correlate with temperature. The results suggest that the toxic effect of Cd on elongation growth of coleoptile segments is connected with the decrease of the PM H(+)-ATPase activity and probably with Cd-induced high acivity of IAA oxidase, whereas the effect of Pb did not depend on activity of any of the enzymes.

Effects of juglone and lawsone on oxidative stress in maize coleoptile cells treated with IAA

Despite the large number of papers that have been published on juglone and lawsone cytotoxicity, little is known about their redox cycling properties in plants. Here, the effects of these two naturally occurring naphthoquinones on H2O2 production and the activity of ROS scavenging enzymes (SOD, POX, CAT) in maize coleoptile segments were measured. It was found that lawsone was a more effective oxidative stress inducer in maize coleoptile cells than juglone. The results suggested that oxidative stress imposed by juglone and lawsone was one of the mechanisms of allelopathic action of the studied quinones in plants.

Effect of K+ and Ca2+ on the indole-3-acetic acid- and fusicoccin-induced growth and membrane potential in maize coleoptile cells

The role of K+ and Ca2+ in plant growth and development is complex and needs extensive physiological studies. Here, the effects of K+ and Ca2+ ions on growth and membrane potential in the presence of either auxin (IAA) or fusicoccin (FC) were studied in maize coleoptiles. The results presented in this article demonstrate that the inhibitory effect of Ca2+ on growth is significantly restored in the presence of K+. This effect is probably due to competitive inhibition of K+ channels by Ca2+ ions, although other possibilities should be taken into consideration.

The effect of auxins (IAA and 4-Cl-IAA) on the redox activity and medium pH of Zea mays L. root segments

Indole-3-acetic acid (IAA) and 4-chloroindole-3-acetic acid (4-Cl-IAA) were tested at different concentrations and times for their capacity to change the redox activity and medium pH of maize root segments. The dose-response surfaces (dose-response curves as a function of time) plotted for redox activity and changes in medium pH (expressed as ΔpH) had a similar shape for both auxins, but differed significantly at the optimal concentrations. With 4-Cl-IAA, the maximal values of redox activity and medium pH changes were observed at 10−10 M, which was a 100-fold lower concentration than with IAA. Correlations were observed between redox activity and medium pH changes at the optimal concentrations of both IAA and 4-Cl-IAA. The results are discussed herein, taking into account both the concentration of the auxins and the effects produced by them.