"ENERGY"-DEPENDENT QUENCHING OF CHLOROPHYLL FLUORESCENCE and THERMAL ENERGY DISSIPATION IN INTACT LEAVES DURING INDUCTION OF PHOTOSYNTHESIS

Abstract— Intact leaves, previously adapted to darkness for a prolonged period of time, were suddenly illuminated with a strong, photosynthetically saturating, white light (ca 1500 μmol m⁻² s^_1), resulting in the rapid establishment of a large energy-dependent chlorophyll fluorescence quenching (q_E) as shown by in vivo fluorescence measurements with a pulse amplitude modulation technique. Two different photothermal methods, photoacoustic spectroscopy and photothermal deflection spectroscopy, were used to monitor thermal deactivation of excited pigments during the dark-light transitions. The in vivo photothermal signals measured with both techniques were shown to remain constant during induction of photosynthesis under high light conditions, suggesting that, in contrast to current hypotheses, energy-dependent quenching q_E is not associated with significant changes in thermal dissipation of absorbed light energy in the chloroplasts. When photosynthesis was induced with a low-intensity modulated light, a noticeable decrease in the heat emission yield was observed resulting from the progressive activation of the competing photochemical processes.     

PHOTOTHERMAL DEFLECTION SPECTROSCOPY OF PHOTOSYNTHETIC PIGMENTS in vivo and in vitro

Abstract— This Technical Note describes the design of a photothermal beam deflection apparatus which allows the easy and rapid measurement of thermal dissipation of absorbed light energy in various photosynthetic materials including whole plant leaves. This system is based on the "mirage effect" in which the refractive index gradient induced in a fluid in contact with the sample, irradiated with an intensity-modulated light, causes the periodic deflection of a laser beam parallel to the sample surface. The deflection of the probe laser beam is detected by a position sensor, the output of which is processed by a lock-in amplifier. Photothermal deflection signals can be monitored in vivo in intact leaves placed in various (liquid or gaseous) environments with a satisfactory signal-to-noise ratio between 100 (in water) and 50 (in air) at low modulation frequencies (ca 30 Hz). It is shown that this new and simple photothermal technique is a very sensitive tool for the measurement of absorption spectra of photosynthetic pigments both in vivo (leaves, algae or chloroplasts) and in model systems (Langmuir-Blodgett and solid films of chlorophyll).     

Light-induced carbon dioxide and oxygen uptake in plant leaves measured by the photoacoustic method.

Leaf discs, enclosed in a photoacoustic (PA) chamber, generate two types of PA gas-uptake signals under certain conditions. Type I is manifested by a severe signal decrease that develops slowly under very low-light intensity and often reaches negative values. It is partially reversed by low-intensity far-red light. Type II occurs transiently in modulated far-red light. It is manifested by a rapid and dramatic decrease of the PA signal, upon the addition of short-wave background light, which is subsequently reversed. It differs from type-I uptake in that it occurs at much higher total light intensities. A thorough study, including modulation frequency and atmospheric composition dependencies, indicates different mechanisms for the two types of uptakes. Type-I uptake results from CO2 accumulation in the PA cell by leaf respiration and reflects modulations in CO2 solubilization. Type-II uptake likely reflects oxygen photoreduction in photosystem I, occurring prior to the activation of photosynthesis (i.e. during photosynthesis induction). This is supported by the complete suppression of type-II uptake when O2 was removed. Also, type-II uptake was only mildly sensitive to CO2 elimination, whereas type-I uptake was totally dependent on the presence of CO2. Type-II uptake consists usually of two uptake waves. Fluorescence transients measured in parallel give further support to the reality and interpretation of these two uptake waves. PA could thus provide a unique opportunity to monitor oxygen photoreduction in vivo with high sensitivity and time resolution.     

ANTAGONISTIC EFFECTS OF RED AND FAR-RED LIGHTS ON THE STABILITY OF PHOTOSYSTEM II IN PEA LEAVES EXPOSED TO HEAT

Abstract— The presence of light during exposure of intact pea leaves to high temperature (40°C) protects Photosystem II (PSII) against inactivation, as indicated by the preservation of the maximal variable 685 nm chlorophyll fluorescence and the photosynthetic oxygen evolution. This photoprotection was observed (i) to be saturated at low fluence rates ( ca 10 W m^-2) and (ii) to be strongly dependent on the spectral characteristics of the light. It was specifically induced by red light (630–670 nm) whereas other wavelengths were much less protective. A strong antagonism between red and far-red lights was also observed, with PSII stabilization by red light being partially cancelled by additional far-red light.     

HEAT- AND LIGHT-INDUCED CHLOROPHYLL a FLUORESCENCE CHANGES IN POTATO LEAVES CONTAINING HIGH OR LOW LEVELS OF THE CAROTENOID ZEAXANTHIN: INDICATIONS OF A REGULATORY EFFECT OF ZEAXANTHIN ON THYLAKOID MEMBRANE FLUIDITY

Potato leaf discs were infiltrated in darkness with a buffer of pH 5 containing 100 M ascorbate, resulting in a massive conversion of the carotenoid violaxanthin to zeaxanthin. In vivo measurements of modulated chlorophyll a fluorescence indicated that this treatment (1) caused a marked upward shift of the threshold temperature at which photosystem II denatures and (2) noticeably inhibited the rate of dark reoxidation of the reduced plastoquinone (at low temperature). These changes were not induced in leaves infiltrated with a buffer of pH 5 containing no ascorbate or with 100 mM ascorbate at pH >7.2. The above-mentioned effects were also observed during heat acclimation (34°C for several days) of potato plants and suggested that zeaxanthin interacts with the lipid phase of the thylakoid membranes. Based on those results and the previous data obtained with model systems, it is suggested that the xanthophyll cycle could be a regulatory mechanism adjusting thylakoid membrane fluidity, the significance of which for the photoprotection of the photosynthetic apparatus is discussed.