Yoshihiro Yamane

Increases in the fluorescence Fo level and reversible inhibition of Photosystem II reaction center by high-temperature treatments in higher plants

Effects of high temperatures on the fluorescence Fm (maximum fluorescence) and Fo (dark level fluorescence) levels were studied and compared with those of the photochemical reactions of PS II. These comparisons were performed during and after the high temperature treatments. The following results were obtained; (1) increases in the Fo level at high temperatures were partly reversible, (2) the Fm level in the presence of dithionite in spinach chloroplasts decreased at high temperatures and also showed a partial reversibility, (3) photoreductions of pheophytin a and Qa were reversibly inhibited at high temperatures parallel to the decrease in the difference between the Fm and Fo levels, and (4) the decrease in the fluorescence Fm level seemed to be related to denaturation of chlorophyll-proteins. All the data suggested that, as well as the separation of light-harvesting chlorophyll a/t b protein complexes of PS II from the PS II core complexes, partly reversible inactivation of the PS II reaction center at high temperatures is the cause of the increase in the Fo level.

Effects of high temperatures on the photosynthetic systems in spinach: Oxygen-evolving activities, fluorescence characteristics and the denaturation process

Activities of oxygen evolution, fluorescence Fv (a variable part of chlorophyll fluorescence) values, and amounts of the 33 kDa protein remaining bound to the thylakoids in intact spinach chloroplasts were measured during and after high-temperature treatment. The following results were obtained. (1) Both the Fv value and the flash-induced oxygen evolution measured by an oxygen electrode were decreased at high temperatures, but they showed partial recovery when the samples were cooled down and incubated at 25°C for 5 min after high-temperature treatment. (2) Oxygen evolution was more sensitive to high temperatures than the Fv value, and the decrease in the Fv/Fm ratio at high temperatures rather corresponded to that in the oxygen evolution measured at 25°C after high-temperature treatment. (3) Photoinactivation of PS II was very rapid at high temperatures, and this seems to be a cause of the difference between the Fv values and the oxygen-evolving activities at high temperatures. (4) At around 40°C, the manganese-stabilizing 33 kDa protein of PS II was supposed to be released from the PS II core complexes during heat treatment and to rebind to the complexes when the samples were cooled down to 25°C. (5) At higher temperatures, the charge separation reaction of PS II was inactivated, and the PS II complexes became less fluorescent, which was recovered partially at 25°C. (6) Increases in the Fv value due to a large decrease in the electron flow from QA to QB became prominent after high-temperature treatment at around 50°C. This was the main cause of the discrepancy between the Fv values and the oxygen-evolving activities measured at 25°C. Relationship between the process of heat inactivation of PS II reaction center complexes and the fluorescence levels is discussed.

Temperature Acclimation of Photosynthesis and Related Changes in Photosystem II Electron Transport in Winter Wheat

Winter wheat (Triticum aestivum L. cv Norin No. 61) was grown at 25°C until the third leaves reached about 10 cm in length and then at 15°C, 25°C, or 35°C until full development of the third leaves (about 1 week at 25°C, but 2–3 weeks at 15°C or 35°C). In the leaves developed at 15°C, 25°C, and 35°C, the optimum temperature for CO2-saturated photosynthesis was 15°C to 20°C, 25°C to 30°C, and 35°C, respectively. The photosystem II (PS II) electron transport, determined either polarographically with isolated thylakoids or by measuring the modulated chlorophyll a fluorescence in leaves, also showed the maximum rate near the temperature at which the leaves had developed. Maximum rates of CO2-saturated photosynthesis and PS II electron transport determined at respective optimum temperatures were the highest in the leaves developed at 25°C and lowest in the leaves developed at 35°C. So were the levels of chlorophyll, photosystem I and PS II, whereas the level of Rubisco decreased with increasing temperature at which the leaves had developed. Kinetic analyses of chlorophyll afluorescence changes and P700 reduction showed that the temperature dependence of electron transport at the plastoquinone and water-oxidation sites was modulated by the temperature at which the leaves had developed. These results indicate that the major factor that contributes to thermal acclimation of photosynthesis in winter wheat is the plastic response of PS II electron transport to environmental temperature.

Reduction of QA in the dark: Another cause of fluorescence Fo increases by high temperatures in higher plants

Increases in the chlorophyll fluorescence Fo (dark level fluorescence) during heat treatments were studied in various higher plants. Besides the dissociation of light-harvesting chlorophyll a/b protein complexes from the reaction center complex of PS II and inactivation of PS II, dark reduction of QA via plastoquinone (PQ) seemed to be related to the Fo increase at high temperatures. In potato leaves or green tobacco cultured cells, a part of the Fo increase was quenched by light, reflecting light-induced oxidation of QA- which had been reduced in the dark at high temperatures. Appearance of the Fo increase due to QA reduction depended on the plant species, and the mechanisms for this are proposed. The reductants seemed to be already present and formed by very brief illumination of the leaves at high temperatures. A ndhB-less mutant of tobacco showed that complex I type NAD(P)H dehydrogenase is not involved in the heat-induced reduction of QA. Quite strong inhibition of the QA reduction by diphenyleneiodonium suggests that a flavoenzyme is one of the electron mediator to PQ from the reductant in the stroma. Reversibility of the heat-induced QA reduction suggests that an enzyme(s) involved is activated at high temperatures and mostly returns to an inactive form at room temperature (25 °C).

Effects of High Temperatures on Photosynthetic Systems: Fluorescence Fo Increases in Cyanobacteria

Oxygen evolution is one of the most heat-sensitive sites in photosynthetic organisms. To investigate effects of heat stress on photosynthesis, high-temperature (HT) -induced fluorescence Fo (minimum fluorescence) increases and Fm (maximum fluorescence) decreases had been studied by many workers (1). In higher plants, the Fo increase was attributed to irreversible detachment of light-harvesting chlorophyll a/b protein complexes from the reaction center (RC) complexes of Photosystem (PS) II, to partly reversible inactivation of PS 11 (2, 3), and to dark reduction of QA (4). The Fm decrease is related to inhibition of oxygen evolution (5, 6). However, Fm levels in the presence of 3- (3,4-dichlorophenyl) -1,1-dimethylurea and NH2OH, which cancelled the inhibitory effect of heat stress on oxygen evolution, were also decreased by HT treatments, showing that RCs of PS II were inactivated by high temperature-stress and that the inactivated PS II RC complexes become less fluorescent (6). In cyanobacteria, however, little is known about the effects of HTs on photosynthetic systems and on these fluorescence parameters. In this report, we show that the Fo increase in cyanobacteria is due to partly reversible release of phycobilisomes from the RC complexes of PS II and to partly reversible inactivation of PS II RC.