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The effects of oligomycin on photosynthesis and respiration in relation to ATP production in chloroplasts and mitochondria were investigated in protoplasts isolated from the detached pea (Pisum sativum L cv. Iłowiecki.) and barley (Hordeum vulgare L. cv. Gunilla) leaves treated 5 mM Pb(NO3)2. The oligomycin (OM), an inhibitor of oxidative phosphorylation at 0.1 pM concentration caused the inhibition of photosynthesis rate in the protoplasts from both the control and the Pb-treated pea leaves. The respiration rate and ATP/ADP ratio in the protoplasts and the activity of ATPase in mitochondria, were also diminished in the control protoplasts. These effects were not observed in the protoplasts and mitochondria isolated from the Pb-treated leaves. Oligomycin, an inhibitor of photophosphorylation at 10 pM concentration decreased ATPase activity in chloroplasts from both the control and the Pb- treated leaves. Using the method of rapid fractionation of barley protoplasts it was shown that the ATP/ADP ratio in the mitochondria from Pb-treated leaves was largely suppressed (from 1.8 to 0.4) by OM under nonphotorespiratory conditions (high CO2), whereas under photorespiratory conditions (low CO2) this ratio was high (5.3) and under OM decreased less (to 3.1). Our results indicate that oligomycin, in organelle isolated from Pb-treated leaves, had no inhibitory effect on the mitochondrial ATPase, whereas it inhibited chloroplasts ATPase. We suggest that Pb ions affected the catalytic cycle and/or conformational changes of ATPase in pea chloroplasts differently than in mitochondria. The differences in Pb responses may reflect fine mechanisms for the regulation of ATP production in the plant cells under stress conditions.
Photosynthesis and transpiration rate of detached leaves of pea (Pisum sativum L. cv. Iłowiecki) exposed to solution of Pb(NO₃)₂ at 1 or 5 mmol·dm⁻³ concentrations were inhibited. The higher concentration of this toxicant decreased photosynthesis and transpiration rates 2 and 3 times respectively, and increased respiration by about 20 %, as measured after 24 hours of treatment. Similarly to Pb(NO₃)₂, glyceraldehyde solution, an inhibitor of phosphoribulokinase, at 50 mmol·dm⁻³ concentration decreased the rates of photosynthesis and transpiration during introduction into pea leaves. The rate of dark respiration, however, remained unchanged during 2 hours of experiment. The potential photochemical efficiency of PS II (Fv/Fm) and the activity of Rubisco (EC 4.1.1.39) at 5 mmol·dm⁻³ of Pb(NO₃)₂ were lowered by 10 % and 20 % respectively, after 24 hours. Neither changes in the activity of PEPC (EC 4.1.1.31) or protein and pigment contents were noted in Pb-treated leaves. The photosynthetic activity of protoplasts isolated from leaves treated for 24 or 48 hours with Pb(NO₃)₂ at 5 mmol·dm⁻³ concentration was decreased 10 % or 25 %, whereas, the rate of dark respiration was stimulated by about 40 % and 75 %, respectively. The content of abscisic acid, a hormone responsible for stomatal closure, in detached pea leaves treated for 24 h with 5 mmol·dm⁻³ of Pb(NO₃)₂ solution was increased by about 3 times; a longer (48h) treatment led to further increase (by about 7 times) in the amount of this hormone. The results of our experiments provide evidences that CO₂ fixation in detached pea leaves, at least up to 24 hours of Pb(NO₃)₂ treatment, was restricted mainly by stomatal closure.
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