Characterization of juvenile and adult leaves of Eucalyptus globulus showing distinct heteroblastic development: photosynthesis and volatile isoprenoids.

Heteroblastic Eucalyptus (Eucalyptus globulus L.) leaves were characterized for their functional diversity examining photosynthesis and photosynthesis limitations, transpiration, and the emission of isoprene and monoterpenes. In vivo and combined analyses of gas-exchange, chlorophyll fluorescence, and light absorbance at 830 nm were made on the adaxial and abaxial sides of juvenile and adult leaves. When adult leaves were reversed to illuminate the abaxial side, photosynthesis and isoprene emission were significantly lower than when the adaxial side was illuminated. Monoterpene emission, however, was independent on the side illuminated and similarly partitioned between the two leaf sides. The abaxial side of adult leaves showed less diffusive resistance to CO(2) acquisition by chloroplasts, but also lower ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity, than the adaxial leaf side. In juvenile leaves, photosynthesis, isoprene, and monoterpene emissions were similar when the adaxial or abaxial side was directly illuminated. In the abaxial side of juvenile leaves, photosynthesis did not match the rates attained by the other leaf types when exposed to elevated CO(2), which suggests the occurrence of a limitation of photosynthesis by ribulose bisphosphate (RuBP) regeneration. Accordingly, a reduced efficiency of both photosystems and a high non-radiative dissipation of energy was observed in the abaxial side of juvenile leaves. During light induction, the adaxial side of juvenile leaves also showed a reduced efficiency of photosystem II and a large non-radiative energy dissipation. Our report reveals distinct functional properties in Eucalyptus leaves. Juvenile leaves invest more carbon in isoprene, but not in monoterpenes, and have a lower water use efficiency than adult leaves. Under steady-state conditions, in adult leaves the isobilateral anatomy does not correspond to an equal functionality of the two sides, while in juvenile leaves the dorsiventral anatomy does not result in functional differences in primary or secondary metabolism in the two sides. However, photochemical limitations may reduce the efficiency of carbon fixation in the light, especially in the abaxial side of juvenile leaves.

Simultaneous analysis of prompt and delayed chlorophyll a fluorescence in leaves during the induction period of dark to light adaptation.

An attempt is made to reveal the relation between the induction curves of delayed fluorescence (DF) registered at 0.35-5.5 ms and the prompt chlorophyll fluorescence (PF). A simple formulation was proposed to link the ratio of the transient values of delayed and variable fluorescence with the redox state of the primary electron acceptor of Photosystem II--QA, and the thylakoid membrane energization. The term luminescence potential (UL) was introduced, defined as the sum of the redox potential of QA and the transmembrane proton gradient. It was shown that UL is proportional to the ratio of DF to the variable part of PF. The theoretical model was verified and demonstrated by analysing induction courses of PF and millisecond DF, simultaneously registered from leaves of barley--wild-type and the chlorophyll b-less mutant chlorina f2. A definitive correlation between PF and DF was established. If the luminescence changes are strictly due to UL, the courses of DF and PF are reciprocal and the millisecond DF curve resembles the first derivative of the PFt function.

High-temperature damage and acclimation of the photosynthetic apparatus : I. Temperature sensitivity of some photosynthetic parameters of chloroplasts isolated from acclimated and non-acclimated bean leaves.

The thermosensitivity of delayed fluorescence, the relative values of variable chlorophyll fluorescence and the degree of quenching of 9-aminoacridine fluorescene were studied in the chloroplasts from heat-acclimated and non-acclimated (treated 6 h at 52,5°C) young bean plants. The temperature sensitivity of each parameter studied was defined by that temperature at which chloroplast activity decreased by 50% (T50) of its maximum value. There was appreciable increase in the thermostability of membrane energization in chloroplasts isolated from acclimated and non-acclimated plants compared with the controls. The photosynthetic parameters differed according to the suspending medium and the preacclimation treatment. When chloroplast were suspended in phosphate buffer with the addition of stabilizing compounds (2 M sucrose or 0.5% human serum albumin) the thermostability of the thylakoid membranes increased, as was evident by the increases in T50 of about 8-10° C (sucrose) and 2-5° C (human serum albumin) for all the parameters investigated. Photoinduced quenching of 9-aminoacridine fluorescence decreased to some extent in the presence of protective compounds, but in chloroplasts from acclimated plants the T50 was practically equal to that for their long-lived luminescence under the same conditions. At the thylakoid membrane level, acclimation was clearly manifested as an increased thermostability of photoinduced proton-gradient formation.

High-temperature damage and acclimation of the photosynthetic apparatus : II. Effect of mono- and divalent cations and pH on the temperature sensitivity of some functional characteristics of chloroplasts isolated from heat-acclimated and non-acclimated bean plants.

The influence of mono- (K(+)) and divalent (Mg(2+)) cations and protons (pH) on the temperature sensitivity of thylakoid membranes was investigated in three groups of young bean plants (control, heat-acclimated and non-acclimated). Thylakoid-membrane function was monitored by second and millisecond delayed fluorescence and 9-aminoacridine fluorescence quenching. It was established that metal ions at investigated concentrations decreased the thermostability of the photosynthetic parameters - an increase of MgSO4 concentration from 0.1 to 20 mM decreased the temperature of their half-inactivation (T50) by 13°C. At the same time the pH dependence of the thermal stability of these parameters showed a maximum at pH 5.5-6.5. The half-inactivation temperatures of those photosynthetic parameters connected with the ability of the thylakoid membrane to form light-induced proton gradients increased by 6-7°C in the heat-acclimated plants compared with the control. It was assumed that the temperature inactivation of photosynthetic electron transfer and the energization of the thylakoid membrane was determined both by the thermoinduced dissociation of the light-harvesting chlorophyll a/b protein complex from PSII, leading to destruction of the excitation energy transfer to the reaction centres, and by the thermal denaturation of the membrane-protein components. The rate of these processes was probably controlled by the size of the negative surface charge and the viscosity of the thylakoid membrane.

Biosynthetic cause of in vivo acquired thermotolerance of photosynthetic light reactions and metabolic responses of chloroplasts to heat stress.

Thermotolerance of photosynthetic light reactions in vivo is correlated with a decrease in the ratio of monogalactosyl diacylglycerol to digalactosyl diacylglycerol and an increased incorporation into thylakoid membranes of saturated digalactosyl diacylglycerol species. Although electron transport remains virtually intact in thermotolerant chloroplasts, thylakoid protein phosphorylation is strongly inhibited. The opposite is shown for thermosensitive chloroplasts in vivo. Heat stress causes reversible and irreversible inactivation of chloroplast protein synthesis in heat-adapted and nonadapted plants, respectively, but doe not greatly affect formation of rapidly turned-over 32 kilodalton proteins of photosystem II. The formation on cytoplasmic ribosomes and import by chloroplasts of thylakoid and stroma proteins remain preserved, although decreased in rate, at supraoptimal temperatures. Thermotolerant chloroplasts accumulate heat shock proteins in the stroma among which 22 kilodalton polypeptides predominate. We suggest that interactions of heat shock proteins with the outer chloroplast envelope membrane might enhance formation of digalactosyl diacylglycerol species. Furthermore, a heat-induced recompartmentalization of the chloroplast matrix that ensures effective transport of ATP from thylakoid membranes towards those sites inside the chloroplast and the cytoplasm where photosynthetically indispensable components and heat shock proteins are being formed is proposed as a metabolic strategy of plant cells to survive and recover from heat stress.