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Fluorescence induction at -196 degrees C has been monitored in chloroplasts rapidly frozen after poising at different redox potentials at room temperature. It was found that, as at room temperature, the initial level of fluorescence observed upon shutter opening (Fo), relative to the final level observed after 10 seconds of illumination (Fm) increased as the redox potential of the chloroplasts was lowered. Redox titration and revealed the presence of two quenching components with Em, 7.8 at -70 mV and -275 mV accounting for approx. 75% and 25% of the variable fluorescence (Fv). Parallel observation of fluorescence yield at room temperature similarly gave two components, with Em, 7.8 at -95 mV and -290 mV, also accounting for appox. 75% and 25%. Simultaneous measurement of fluorescence emission at -196 degrees C at 695 nm and 735 nm indicated that both emissions are quenched by the same redox components.

The effects of low temperature on the synthesis and stability of the 32 kDa D1 protein of photosystem II were invesligated in chioroplasts isolated from maize (Zea mays cv. LG11) leaves. The synthesis of D1 by intact chloroplasts in vitro was strongly dependent on temperature; the Q10 for the initial rate of incorporation of [35S]-methionine into D1 was ca. 2.6 over the range 13-25 °C. The synthesis of other thylakoid polypeptides exhibited a similar temperature dependence, whilst synthesis of stromal proteins was considerably less temperature-dependent, with the exception of two polypeptides of ca. 56 and 59.5 kDa. The stability of newly-synthesized D1 in the thylakoid membranes was dependent both on the temperature at which the plants were grown and on the temperature during the pulse-labelling period when the protein was synthesized. In chloroplasts isolated from maize leaves grown at 25°C, D1 that was synthesized and assembled at 25°C in vitro was rapidly degraded during the chase period. At lower chase temperatures the protein was more stable. When chloroplasts from 25°C-grown leaves were pulse-labelled at 13°C, the stability of D1 was markedly enhanced at all temperatures during the chase period. This effect was even more pronounced in chloroplasts isolated from plants grown at 14°C. The implications of these results are discussed with regard to the ability of maize to recover from photoinhibitory damage at low temperatures. [ABSTRACT FROM AUTHOR]

Tissue-specific effects of low growth temperature on maize chloroplast thylakoid protein accumulation were analysed using immunocytology. Sections of leaves from plants grown at 25 and 14°C were probed with antibodies to specific chloroplast thylakoid proteins from the four major protein multisubunit complexes of the thylakoid membrane followed by fluorescein-conjugated goat anti-rabbit antibodies. At a normal growth temperature of 25°C, the 32 kDa D1 protein of the photosystem II reaction centre and the 33 kDa protein of the extrinsic oxygen- evolving complex of photosystem II are both accumulated to a greater degree in the mesophyll than in the bundle sheath chloroplasts. In contrast, subunit II of photosystem I, cytochrome f and the α and β-subunits of ATP synthetase are predominant in the bundle sheath thylakoids at 25 °C. A striking difference between the 25 °C-grown and the 14°C-grown leaf tissue was the presence in the latter of (20-30%) cells whose chloroplasts apparently completely lack several of the thylakoid proteins. In plants grown at 14°C, the accumulation of the 33 kDa protein of the extrinsic oxygen-evolving complex of photosystem II was apparently unchanged, but other thylakoid proteins showed a significant reduction, The uneven distribution of proteins between the bundle sheath and mesophyll chloroplasts observed at 25°C was also maintained at 14 °C. Reduction in the fluorescence at 14°C was manifested either as an overall reduction in the diffuse fluorescence across the chloroplast profiles or less frequently as a reduction to small discrete bodies of intense fluorescence. The significance of these results to low-temperature-induced reduction in the photosynthetic productivity of maize is discussed. [ABSTRACT FROM AUTHOR]

The kinetics of a range of chlorophyll fluorescence parameters, non-cyclic electron transport and the capacity of the thylakodis to bind Atrazine were examined during photoinhibition treatment of intact pea chloroplasts. Parameters of fluorescence induction of chloroplasts in the presence and absence of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea at 20°C and at 77 K were determined. The contributions of photochemical and non-photochemical quenching processes to the loss of fluorescence during photoinhibitory treatment were assessed. Two distinct phases of photoinhibitory damage were observed. During the initial 5 min period of exposure to light the minimal fluorescence level (F0) increased, whilst the maximal fluorescence level (FP)decreased, both coupled and uncoupled non-cyclic electron transport to methyl viologen decreased and the ability to bind Atrazine to the thylakoids decreased. Fluorescence analyses demonstrated that during this period thylakoids were becoming increasingly less efficient at generating and maintaining a transmembrane proton electrochemical gradient. Photoinhibitory damage that occurred at later times between 5 and 20 min was of a very different nature. Both (F0) and (FP) declined, a loss of coupled and uncoupled non-cyclic electron transport was observed together with a loss of the capacity to photo-oxidize water. However, no further loss of Atrazine-binding was associated with such changes. A consistent decrease in the quantum yield of non-cyclic electron transport was also observed throughout photoinhibition treatment. The possibility of two distinct mechanisms of photoinhibitory damage to the photosynthetic apparatus is discussed. [ABSTRACT FROM AUTHOR]

Photosynthetic electron transport activities and the ability to generate and maintain a trans-thylakoid proton electrochemical gradient were examined during chloroplast development in 4-day-old wheat leaves grown under a diurnal light regime. Polarographic and spectrophotometric studies on leaf tissue demonstrated that poorly developed chloroplasts at the leaf base could photo-oxidize water and transfer electrons from photosystem 2 to photosystem 1. The capacity for non-cyclic whole-chain electron transport increased during chloroplast development. Thylakoids isolated from the leaf base, although capable of pumping protons into the intrathylakoid space, could not maintain a transmembrane proton electrochemical gradient; this ability developed at later stages of chloroplast biogenesis in the leaf. The implications of these results for the energies of the developing leaf are discussed. [ABSTRACT FROM AUTHOR]

The effect of growth temperatures on quantum yield (φ) was examined for leaves at different stages of development within the immature canopies of two crops of field grown maize (Zea mays cv. LG11) sown on 3 May and 20 June 1990. During the period of 23 to 49d after sowing, the crop sown on the 3 May experienced temperatures below 10°C on 19 occasions compared with only two for the crop sown on 20 June. A period of severe chilling at the end of May and the beginning of June was associated with a marked reduction in φ for all leaves in the early-sown crop. This chill-induced depression in φ was greater in recently emerged than more mature leaves in the canopy and was found to be accompanied by modifications in the polypeptide profiles of thylakoids isolated from the leaves. During the chilling period, decreases in some polypeptides, notably in the range of 41-42 and 20 kDa apparent molecular size, and increases of polypeptides of c. 15-16 kDa were observed compared with leaves developing at warmer temperatures in July. The efficiency of converting intercepted radiation into dry matter (conversion efficiency) was 42% lower in the early- than late-sown crop, but no significant relationship between conversion efficiency and quantum yield was found in either treatment. [ABSTRACT FROM AUTHOR]