Recovery of photosynthesis and phtosystem II fluorescence in Chlamydomonas reinhardtii after exposure to three levels of high light

Recovery from 60 min of photoinhibitory treatment at photosynthetic photon flux densities of 500, 1400 and 2200 μMmol m^?2 s^? was followed in cells of the green alga Chlamydomonas reinhardtii grown at 125 μMmol m^?2 s^?1. These light treatments represent photoregulation, moderate photoinhibition and strong photoinhibition, respectively. Treatment in photoregulatory light resulted in an increased maximal rate of oxygen evolution (P_max) and an increased quantum yield (Φ), but a 15% decrease in F_v/F_M. Treatment at moderately photoinhibitory light resulted in a 30% decrease in F_v/F_M and an approximately equal decrease in Φ. Recovery in dim light restored F_v/F_M within 15 and 45 min after high light treatment at 500 and 1400 μMmol m^?2 s^?1, respectively. Convexity (Θ), a measure of the extent of co-limitation between PS II turnover and whole-chain electron transport, and Φ approached, but did not reach the control level during recovery after exposure to 1400 μMmol m^?2 s^?1, whereas P_max increased above the control. Treatment at 2200 μMmol m^?2 s^?1 resulted in a strong reduction of the modeled parameters Φ, Θ and P_max. Subsequent recovery was initially rapid but the rate decreased, and a complete recovery was not reached within 120 min. Based on the results, it is hypothesized that exposure to high light results in two phenomena. The first, expressed at all three light intensities, involves redistribution within the different aspects of PS II heterogeneity rather than a photoinhibitory destruction of PS II reaction centers. The second, most strongly expressed at 2200 μmol m^?2 s^?1, is a physical damage to PS II shown as an almost total loss of PS II_α and PS II Q_B-reducing centers. Thus recovery displayed two phase, the first was rapid and the only visible phase in algae exposed to 500 and 1400 μmol m^?2 s^?1. The second phase was slow and visible only in the later part of recovery in cells exposed to 2200 μmol m^?2 s^?1.     

The effects of photoinhibition on the photosynthetic light-response curve of green plant cells (Chlamydomonas reinhardtii)

The photosynthetic response to light can be accurately defined in terms of (1) the initial slope (quantum yield); (2) the asymptote (light-saturated rate); (3) the convexity (rate of bending); and (4) the intercept (dark respiration). The effects of photoinhibition [which damages the reaction centre of photosystem II (PSII)] on these four parameters were measured in optically thin cultures of green plant cells (Chlamydomonas reinhardtii). The convexity of the light-response curve decreased steadily from a value of 0.98 (indicating a sharply bending response) to zero (indicating Michaelis-Menten kinetics) in response to increasing photoinhibition. Photoinhibition was quantified from the quantum yield of inhibited cells relative to that of control cells. The quantum yield was estimated by applying linear regression to low-light data or by fitting a non-rectangular hyperbola. Assuming the initial slope is linear allowed comparison with earlier work. However, as the convexity was lowered this assumption resulted in a significant underestimate of the true quantum yield. Thus, the apparent level of photoinhibition required for a zero convexity and the initial decrease in light-saturated photosynthesis depended upon how the quantum yield was estimated. If the initial slope of the light response was assumed to be linear the critical level of inhibition was 60%. If the linear assumption was not made, the critical level was 40%. At the level of inhibition where the convexity reached zero, the light-saturated rate of photosynthesis also began to decrease, indicating that this level of inhibition caused photosynthesis to be limited at all light intensities by the rate of PSII electron transport. At this level of inhibition the F_m-F_i signal (where F_m is maximal chlorophyll fluorescence and F_i is intermediate chlorophyll fluorescence of dark adapted cells; Briantais et al. 1988) from the fluorescence induction curve was zero and the F_i-F_o signal (where F_o is initial chlorophyll fluorescence of dark adapted cells) was 30% of the control, indicating dramatic reduction or complete elimination of one type of PSII. These data do not contradict published mathematical models showing that the ratio of the maximum speed of electron transport in PSII relative to the maximum speed of plastoquinone electron transport can determine the convexity of the photosynthetic response to light.Abbreviations and Symbols Chl chlorophyll content - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_o, F_i, F_m initial, intermediate, and maximal Chl fluorescence of dark adapted cells - P rate of net photosynthesis per unit chlorophyll (mol-(mg Chl)^–1 · s^–1) - PSII photosystem II - PQ plastoquinone - initial slope to the light-response curve - convexity (rate of bending) of the light-response curve of photosynthesis - Q photosynthetically active photon flux density (400–700 nm, mol · m^–2 · ^–1) The present investigation was supported by the Swedish Council for Forestry and Agricultural Research, the Swedish Environmental Protection Board, and the Swedish Natural Science Research Council. We thank Dr. Deborah D. Kaska (Department of Biological Sciences, University of California, Santa Barbara, Calif., USA) for giving us Chlamydomonas algae. We thank Professor G. Öquist (Department of Plant Physiology, University of Umea, Umea, Sweden) for his encouragement, valuable comments and discussion.     

Changes in Photosystem II fluorescence in Chlamydomonas reinhardtii exposed to increasing levels of irradiance in relationship to the photosynthetic response to light

The effects of a 60 min exposure to photosynthetic photon flux densities ranging from 300 to 2200 mol m^–2s^–1 on the photosynthetic light response curve and on PS II heterogeneity as reflected in chlorophyll a fluorescence were investigated using the unicellular green alga Chlamydomonas reinhardtii. It was established that exposure to high light acts at three different regulatory or inhibitory levels; 1) regulation occurs from 300 to 780 mol m^–2s^–1 where total amount of PS II centers and the shape of the light response curve is not significantly changed, 2) a first photoinhibitory range above 780 up to 1600 mol m^–2s^–1 where a progressive inhibition of the quantum yield and the rate of bending (convexity) of the light response curve can be related to the loss of Q_B-reducing centers and 3) a second photoinhibitory range above 1600 mol m^–2s^–1 where the rate of light saturated photosynthesis also decreases and convexity reaches zero. This was related to a particularly large decrease in PS II centers and a large increase in spill-over in energy to PS I.Abbreviations Chl chlorophyll - DCMU 3,(3,4-dichlorophenyl)-1,1-dimethylurea - F_M maximal fluorescence yield - F_pl intermediate fluorescence yield plateau level - F₀ non-variable fluorescence yield - F_v total variable fluorescence yield (F_M-F₀) - initial slope to the light response curve, used as an estimate of initial quantum yield - convexity (rate of bending) of the light response curve of photosynthesis - LHC light-harvesting complex - P_max maximum rate of photosynthesis - PQ plastoquinone - Q photosynthetically active photon flux density (400–700 nm, mol m^–2s^–1) - PS photosystem - Q_A and Q_B primary and secondary quinone electron acceptor of PS II     

Photosynthesis and photoinhibition in leaves of chlorophyll b-less barley in relation to absorbed light

The response of photosynthesis to absorbed light by intact leaves of wild-type ( Hordeum vulgare L. cv. Gunilla) and chlorophyll b -less barley ( H. vulgare L. cv. Dornaria, chlorina-f2²⁸⁰⁰) was measured in a light integrating sphere. Up to the section where the light response curve bends most sharply the responses of the b -less and wild-type barley were similar but not identical. Average quantum yield and convexity for the mutant light response curves were 0.89 and 0.90, respectively, times those of the wild-type barley. The maximum quantum yield for PSII photochemistry was also 10% lower as indicated by fluorescence induction kinetics (F_v/F_m). Just above the region where the light curve bends most sharply, photosynthesis decreased with time in the mutant but not in the wild-type barley. This decrease was associated with a decrease in F_v/F_m indicating photoinhibition of PSII. This photoinhibition occurred in the same region of the light response curve where zeaxanthin formation occurs. Zeaxanthin formation occurred in both the chlorophyll b -less and wild-type leaves. However, the epoxidation state was lower in the mutant than in the wild-type barley. The results indicate that chlorophyll b -less mutants will have reduced photosynthetic production as a result of an increased sensitivity to photoinhibition and possibly a lowered quantum yield and convexity in the absence of photoinhibition.     

A note on monotonicity assumptions for exact unconditional tests in binary matched-pairs designs

Summary Exact unconditional tests have been widely applied to test the difference between two probabilities for 2 × 2 matched‐pairs binary data with small sample size. In this context, Lloyd (2008, Biometrics 64 , 716–723) proposed an E + M p ‐value, that showed better performance than the existing M p ‐value and C p ‐value. However, the analytical calculation of the E + M p ‐value requires that the Barnard convexity condition be satisfied; this can be challenging to prove theoretically. In this article, by a simple reformulation, we show that a weaker condition, conditional monotonicity, is sufficient to calculate all three p ‐values (M, C , and E + M ) and their corresponding exact sizes. Moreover, this conditional monotonicity condition is applicable to noninferiority tests.     

The single-process biochemical reaction of Rubisco: a unified theory and model with the effects of irradiance, CO₂ and rate-limiting step on the kinetics of C₃ and C₄ photosynthesis from gas exchange

Photosynthesis is the origin of oxygenic life on the planet, and its models are the core of all models of plant biology, agriculture, environmental quality and global climate change. A theory is presented here, based on single process biochemical reactions of Rubisco, recognizing that: In the light, Rubisco activase helps separate Rubisco from the stored ribulose-1,5-bisphosphate (RuBP), activates Rubisco with carbamylation and addition of Mg²⁺, and then produces two products, in two steps: (Step 1) Reaction of Rubisco with RuBP produces a Rubisco-enediol complex, which is the carboxylase-oxygenase enzyme (Enco) and (Step 2) Enco captures CO₂ and/or O₂ and produces intermediate products leading to production and release of 3-phosphoglycerate (PGA) and Rubisco. PGA interactively controls (1) the carboxylation-oxygenation, (2) electron transport, and (3) triosephosphate pathway of the Calvin-Benson cycle that leads to the release of glucose and regeneration of RuBP. Initially, the total enzyme participates in the two steps of the reaction transitionally and its rate follows Michaelis-Menten kinetics. But, for a continuous steady state, Rubisco must be divided into two concurrently active segments for the two steps. This causes a deviation of the steady state from the transitional rate. Kinetic models are developed that integrate the transitional and the steady state reactions. They are tested and successfully validated with verifiable experimental data. The single-process theory is compared to the widely used two-process theory of Farquhar et al. (1980. Planta 149, 78-90), which assumes that the carboxylation rate is either Rubisco-limited at low CO₂ levels such as CO₂ compensation point, or RuBP regeneration-limited at high CO₂. Since the photosynthesis rate cannot increase beyond the two-process theory's Rubisco limit at the CO₂ compensation point, net photosynthesis cannot increase above zero in daylight, and since there is always respiration at night, it leads to progressively negative daily CO₂ fixation with no possibility of oxygenic life on the planet. The Rubisco-limited theory at low CO₂ also contradicts all experimental evidence for low substrate reactions, and for all known enzymes, Rubisco included.     

Variation in tree growth,mortality and recruitment among topographic positions in a warm temperate forest

Questions: Do the population dynamics of trees differ among topographic positions and, if so, how does topographic position affect the population dynamics of species that are distributed in a topography‐specific manner? Which is the most important life stage in determining vegetation patterns? Location: Primary and secondary warm temperate evergreen broad‐leaved forest (40 ‐ 280 m a.s.l.) on the western part of Yakushima Island, Japan. Methods: Mortality, recruitment, DBH growth and distribution of stems (= 5 cm DBH) in a 2.62‐ha plot were surveyed in 1992 and 2002 to determine the relationships between population parameters and (1) topography and (2) distribution patterns of 17 common tree species. Results: Common species (n = 17) were classified into three distribution pattern groups: group A, distributed mainly on convex slopes; group B, on concave slopes, and group C, not aggregated with respect to topographic position. Stem mortality, recruitment and DBH growth were greater in group A than in group B within each topographic class. The hierarchy of stem mortality among topographic classes for groups A and B was convex > planar > concave. Stem recruitment density was relatively high on the convex and concave slopes, respectively, for groups A and B. Conclusions The topographical positions of adult trees were not always most suited for adult survival and growth. For group A, the distribution pattern of adults was determined in the juvenile stage, while this was not the case for group B. Studies of juvenile stages are important for understanding the demographic basis of vegetation distribution patterns.