Wah Soon Chow

The grand design of photosynthesis : acclimation of the photosynthetic apparatus to environmental cues

Dynamic acclimation of the photosynthetic apparatus in response to environmental cues, particularly light quantity and quality, is a widely-observed and important phenomenon which contributes to the tolerance of plants against stress and helps to maintain, as far as possible, optimal photosynthetic efficiency and resource utilization. This mini-review represents a scrutiny of a number of possible photoreceptors (including the two photosystems acting as light sensors) and signal transducers that may be involved in producing acclimation responses. We suggest that regulation by signal transduction may be effected at each of several possible points, and that there are multiple regulatory mechanisms for photosynthetic acclimation.

UV-B damage and protection at the molecular level in plants

Influx of solar UV-B radiation (280–320 nm) will probably increase in the future due to depletion of stratospheric ozone. In plants, there are several targets for the deleterious UV-B radiation, especially the chloroplast. This review summarizes the early effects and responses of low doses of UV-B at the molecular level. The DNA molecules of the plant cells are damaged by UV due to the formation of different photoproducts, such as pyrimidine dimers, which in turn can be combatted by specialized photoreactivating enzyme systems. In the chloroplast, the integrity of the thylakoid membrane seems to be much more sensitive than the activities of the photosynthetic components bound within. However, the decrease of mRNA transcripts for the photosynthetic complexes and other chloroplast proteins are among very early events of UV-B damage, as well as protein synthesis. Other genes, encoding defence-related enzymes, e.g., of the flavonoid biosynthetic pathway, are rapidly up-regulated after commencement of UV-B exposure. Some of the cis-acting nucleotide elements and trans-acting protein factors needed to regulate the UV-induced expression of the parsley chalcone synthase gene are known.

Photoinhibition of photosynthesis represents a mechanism for the long-term regulation of photosystem II

The obligate shade plant, Tradescantia albiflora Kunth grown at 50 μmol photons · m−2 s−1 and Pisum sativum L. acclimated to two photon fluence rates, 50 and 300 μmol · m−2 · s−1, were exposed to photoinhibitory light conditions of 1700 μmol · m−2 · s−1 for 4 h at 22° C. Photosynthesis was assayed by measurement of CO2-saturated O2 evolution, and photosystem II (PSII) was assayed using modulated chlorophyll fluorescence and flash-yield determinations of functional reaction centres. Tradescantia was most sensitive to photoinhibition, while pea grown at 300 μmol · m−2 · s−1 was most resistant, with pea grown at 50 μmol · m−2 · s−1 showing an intermediate sensitivity. A very good correlation was found between the decrease of functional PSII reaction centres and both the inhibition of photosynthesis and PSII photochemistry. Photoinhibition caused a decline in the maximum quantum yield for PSII electron transport as determined by the product of photochemical quenching (qp) and the yield of open PSII reaction centres as given by the steady-state fluorescence ratio, F′vF′m, according to Genty et al. (1989, Biochim. Biophys. Acta 990, 81–92). The decrease in the quantum yield for PSII electron transport was fully accounted for by a decrease in F′vF′m, since qp at a given photon fluence rate was similar for photoinhibited and noninhibited plants. Under lightsaturating conditions, the quantum yield of PSII electron transport was similar in photoinhibited and noninhibited plants. The data give support for the view that photoinhibition of the reaction centres of PSII represents a stable, long-term, down-regulation of photochemistry, which occurs in plants under sustained high-light conditions, and replaces part of the regulation usually exerted by the transthylakoid ΔpH gradient. Furthermore, by investigating the susceptibility of differently lightacclimated sun and shade species to photoinhibition in relation to qp, i.e. the fraction of open-to-closed PSII reaction centres, we also show that irrespective of light acclimation, plants become susceptible to photoinhibition when the majority of their PSII reaction centres are still open (i.e. primary quinone acceptor oxidized). Photoinhibition appears to be an unavoidable consequence of PSII function when light causes sustained closure of more than 40% of PSII reaction centres.

Effects of supplementary ultraviolet-B radiation on photosynthesis in Pisum sativum

Pea plants (Pisum sativum L., cv. Greenfeast) were exposed to supplementary UV-B light (up to 8 days) starting on the 17th day after sowing. The effects of this exposure on photosynthesis and the content and activities of some chloroplast components of the mature leaves of these plants were studied. (i) The total chorophyll content of pea leaves was approximately 40% of that in the control leaves on the 8th day of UV-B exposure. Chlorophyll a levels decreased to a greater extent than the content of chlorophyll b. The decrease in carotenoids paralleled the decrease in chlorophyll b. (ii) On a chlorophyll basis, the contents of Photosystem I and cytochrome f were stable, whereas Photosystem II, ATP hydrolysis by the ATP synthase and the maximum ribulose-1,5-bisphosphate carboxylase (Rubisco) activity decreased by 55, 47 and 80%, respectively, when compared with the controls at the end of the 8-day illumination period. (iii) On a leaf-area basis, Photosystem I and cytochrome f content decreased by 58%, Photosystem II by 80%, ATP hydrolysis by 80%, and Rubisco activity by 90%, when compared with the controls. The in vivo activation of Rubisco was markedly increased in UV-B-treated pea leaves. The underlying mechanisms for these results are discussed.

Green light drives leaf photosynthesis more efficiently than red light in strong white light: Revisiting the enigmatic question of why leaves are green.

The literature and our present examinations indicate that the intra-leaf light absorption profile is in most cases steeper than the photosynthetic capacity profile. In strong white light, therefore, the quantum yield of photosynthesis would be lower in the upper chloroplasts, located near the illuminated surface, than that in the lower chloroplasts. Because green light can penetrate further into the leaf than red or blue light, in strong white light, any additional green light absorbed by the lower chloroplasts would increase leaf photosynthesis to a greater extent than would additional red or blue light. Based on the assessment of effects of the additional monochromatic light on leaf photosynthesis, we developed the differential quantum yield method that quantifies efficiency of any monochromatic light in white light. Application of this method to sunflower leaves clearly showed that, in moderate to strong white light, green light drove photosynthesis more effectively than red light. The green leaf should have a considerable volume of chloroplasts to accommodate the inefficient carboxylation enzyme, Rubisco, and deliver appropriate light to all the chloroplasts. By using chlorophylls that absorb green light weakly, modifying mesophyll structure and adjusting the Rubisco/chlorophyll ratio, the leaf appears to satisfy two somewhat conflicting requirements: to increase the absorptance of photosynthetically active radiation, and to drive photosynthesis efficiently in all the chloroplasts. We also discuss some serious problems that are caused by neglecting these intra-leaf profiles when estimating whole leaf electron transport rates and assessing photoinhibition by fluorescence techniques.

Mechanistic differences in photoinhibition of sun and shade plants

Leaf discs of the shade plant Tradescantia albiflora Kunth grown at 50 μmol · m−2 · s−1, and the facultative sun/shade plant Pisum sativum L. grown at 50 or 300 μmol · m−2, s−1, were photoinhibited for 4 h in 1700 μmol photons m−2 · s−1 at 22° C. The effects of photoinhibition on the following parameters were studied: i) photosystem II (PSII) function; ii) amount of D1 protein in the PSII reaction centre; iii) dependence of photoinhibition and its recovery on chloroplast-encoded protein synthesis; and, iv) the sensitivity of photosynthesis to photoinhibition in the presence or absence of the carotenoid zeaxanthin. We show that: i) despite different sensitivities to photoinhibition, photoinhibition in all three plants occurred at the reaction centre of PSII; ii) there was no correlation between the extent of photoinhibition and the degradation of the D1 protein; iii) the susceptibility to photoinhibition by blockage of chloroplas-tencoded protein synthesis was much less in shade plants than in plants acclimated to higher light; and iv) inhibition of zeaxanthin formation increased the sensitivity to photoinhibition in pea, but not in the shade plant Tradescantia. We suggest that there are mechanistic differences in photoinhibition of sun and shade plants. In sun plants, an active repair cycle of PSII replaces photoinhibited reaction centres with photochemically active ones, thereby conferring partial protection against photoinhibition. However, in shade plants, this repair cycle is less important for protection against photoinhibition; instead, photoinhibited PSII reaction centres may confer, as they accumulate, increased protection of the remaining connected, functional PSII centres by controlled, nonphotochemical dissipation of excess excitation energy.

Photoprotection in a zeaxanthin- and lutein-deficient double mutant of Arabidopsis

When light absorption by a plant exceeds its capacity for light utilization, photosynthetic light harvesting is rapidly downregulated by photoprotective thermal dissipation, which is measured as nonphotochemical quenching of chlorophyll fluorescence (NPQ). To address the involvement of specific xanthophyll pigments in NPQ, we have analyzed mutants affecting xanthophyll metabolism in Arabidopsis thaliana. An npq1 lut2 double mutant was constructed, which lacks both zeaxanthin and lutein due to defects in the violaxanthin de-epoxidase and lycopene ∈-cyclase genes. The npq1 lut2 strain had normal Photosystem II efficiency and nearly wild-type concentrations of functional Photosystem II reaction centers, but the rapidly reversible component of NPQ was completely inhibited. Despite the defects in xanthophyll composition and NPQ, the npq1 lut2 mutant exhibited a remarkable ability to tolerate high light.

Photoprotection and Photoinhibitory Damage

Publisher Summary This chapter discusses the biochemical photoprotection strategies that are adopted by photosynthetic organisms in response to high light. The molecular processes of photosynthesis to withstand photoinhibitory damage include (1) the mechanisms whereby excess excitation energy is dissipated or utilized; (2) the damage to the redox sites when defence strategies fail for one reason or another; and (3) the mechanisms of subsequent repair required to sustain function. The nature of photoinhibitory damage that results when photoprotective strategies fail is also discussed in the chapter. The chapter discusses the recovery from photoinhibitory damage. This review represents a selection of aspects of a fundamental dilemma among photosynthetic organisms: (1) how to maximize the efficiency of light capture and utilization in low light and (2) how to avoid the effects of too much light. There is still a great deal to be learned about photoprotection and photoinhibitory damage. The exact function of the violaxanthin cycle in controlled heat dissipation needs to be clarified.

Light inactivation of functional photosystem II in leaves of peas grown in moderate light depends on photon exposure

To determine the dependence of in vivo photosystem (PS) II function on photon exposure and to assign the relative importance of some photoprotective strategies of PSII against excess light, the maximal photochemical efficiency of PSII (Fv/Fm) and the content of functional PSII complexes (measured by repetitive flash yield of oxygen evolution) were determined in leaves of pea (Pisum satlvum L.) grown in moderate light. The modulation of PSII functionality in vivo was induced by varying either the duration (from 0 to 3 h) of light treatment (fixed at 1200 or 1800 μmol photons · m-2 · s-1) or irradiance (from 0 to 3000 μmol photons · m-2 · s-1) at a fixed duration (1 h) after infiltration of leaves with water (control), lincomycin (an inhibitor of chloroplast-encoded protein synthesis), nigericin (an uncoupler), or dithiothreitol (an inhibitor of the xanthophyll cycle) through the cut petioles of leaves of 22 to 24-day-old plants. We observed a reciprocity of irradiance and duration of illumination for PSII function, demonstrating that inactivation of functional PSII depends on the total number of photons absorbed, not on the rate of photon absorption. The Fv/Fm ratios from photoinhibitory light-treated leaves, with or without inhibitors, declined pseudo-linearly with photon exposure. The number of functional PSII complexes declined multiphasically with increasing photon exposure, in the following decreasing order of inhibitor effect: lincomycin > nigericin > DTT, indicating the central role of D1 protein turnover. While functional PSII and Fv/Fm ratio showed a linear relationship under high photon exposure conditions, in inhibitor-treated leaves the Fv/Fm ratio failed to reveal the loss of up to 25% of the total functional PSII under low photon exposure. The loss of this 25% of less-stable functional PSII was accompanied by a decrease of excitation-energy trapping capacity at the reaction centre of PSII (revealed by the fluorescence parameter, 1/Fo-1/Fm, where Fo and Fm stand for chlorophyll fluorescence when PSII reaction centres are open and closed, respectively), but not by a loss of excitation energy at the antenna (revealed by the fluorescence parameter, 1/Fm). We conclude that (i) PSII is an intrinsic photon counter under photoinhibitory conditions, (ii) PSII functionality is mainly regulated by D1 protein turnover, and to a lesser extent, by events mediated via the transthylakoid pH gradient, and (iii) peas exhibit PSII heterogeneity in terms of functional stability during photon exposure.

Photoinactivation of photosystem II complexes and photoprotection by non-functional neighbours in Capsicum annuum L. leaves

Abstract. Leaf segments from Capsicum annuum plants grown at 100 μmol photons  m−2 s−1 (low light) or 500 μmol photons  m−2 s−1 (high light) were illuminated at three irradiances and three temperatures for several hours. At various times, the remaining fraction (f) of functional photosystem II (PS II) complexes was measured by a chlorophyll fluorescence parameter (1/Fo− 1/Fm, where Fo and Fm are the fluorescence yields corresponding to open and closed PS II traps, respectively), which was in turn calibrated by the oxygen yield per saturating single-turnover flash. During illumination of leaf segments in the presence of lincomycin, an inhibitor of chloroplast-encoded protein synthesis, the decline of f from 1.0 to about 0.3 was mono-exponential. Thereafter, f declined much more slowly, the remaining fraction (≈0.2) being able to survive prolonged illumination. The results can be interpreted as being in support of the hypothesis that photoinactivated PS II complexes photoprotect functional neighbours (G. Öquist et al. 1992, Planta 186: 450–460), provided it is assumed that a photoinactivated PS II is initially only a weak quencher of excitation energy, but becomes a much stronger quencher during prolonged illumination when a substantial fraction of PS II complexes has also been photoinactivated. In the absence of lincomycin, photoinactivation and repair of PS II occur in parallel, allowing f to reach a steady-state value that is determined by the treatment irradiance, temperature and growth irradiance. The results obtained in the presence and absence of lincomycin are analysed according to a simple kinetic model which formally incorporates a conversion from weak to strong quenchers, yielding the rate coefficients of photoinactivation and of repair for various conditions, as well as gaining an insight into the influence of f on the rate coefficient of photoinactivation. They demonstrate that the method is a convenient alternative to the use of radiolabelled amino acids for quantifying photoinactivation and repair of PS II in leaves.