D1 Protein Of PSII Research Articles

Effect of Acclimation to High Temperatures on the Mechanisms of Drought Tolerance in Species with Different Types of Photosynthesis: Sedobassia sedoides (C3–C4) and Bassia prostrata (C4-NADP)

The effect of drought on the morphophysiological, biochemical, and molecular genetic parameters of plants Sedobassia sedoides (Pall.) Freitag G. Kadereit with an intermediate C3–C4-type of photosynthesis and Bassia prostrata (L.) A.J. Scott with a C4-NADP type of photosynthesis grown at different temperatures (25 and 30°C) was studied. A decrease in the biomass, water content, and effective quantum yield (ΦPSII) of PSII, as well as an increase in the expression of the psbA gene encoding the PSII D1 protein under the action of drought, was observed in both species regardless of the growing temperature. Both species showed a decrease in the content of photosynthetic enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC) under drought conditions at 25°С, which was accompanied by a significant increase in the expression of the rbcL and PPDK genes in S. sedoides. Acclimation of S. sedoides plants to elevated temperatures led to an increase in the activity of cyclic electron transport around PSI, to mitigation of the negative effect of drought on the light reactions of photosynthesis (reduction in NPQ) and the content of the PEPC enzyme, as well as to a shift in the ionic balance caused by a decrease in the potassium content. B. prostrata showed greater drought resistance and was characterized by greater thermolability of photosynthetic enzymes, changes in the content and ratio of which allowed this species to maintain growth in drought conditions at different temperatures.

Photosystems under high light stress: throwing light on mechanism and adaptation

High light stress decreases the photosynthetic rate in plants due to photooxidative damage to photosynthetic apparatus, photoinhibition of PSII, and/or damage to PSI. The dissipation of excess energy by nonphotochemical quenching and degradation of the D1 protein of PSII and its repair cycle help against photooxidative damage. Light stress also activates stress-responsive nuclear genes through the accumulation of phosphonucleotide-3'-phosphoadenosine- 5'-phosphate, methylerythritol cyclodiphosphate, and reactive oxygen species which comprise the chloroplast retrograde signaling pathway. Additionally, hormones, such as abscisic acid, cytokinin, brassinosteroids, and gibberellins, play a role in acclimation to light fluctuations. Several alternate electron flow mechanisms, which offset the excess of electrons, include activation of plastid or plastoquinol terminal oxidase, cytochrome b6/f complex, cyclic electron flow through PSI, Mehler ascorbate peroxidase pathway or water-water cycle, mitochondrial alternative oxidase pathway, and photorespiration. In this review, we provided insights into high light stress-mediated damage to photosynthetic apparatus and strategies to mitigate the damage by decreasing antennae size, enhancing NPQ through the introduction of mutants, expression of algal proteins to improve photosynthetic rates and engineering ATP synthase.

Nitric oxide in co-ordination with nitrogen reverses cadmium-inhibited photosynthetic activity by interacting with ethylene synthesis, strengthening the antioxidant system, and nitrogen and sulfur assimilation in mustard (Brassica juncea L.)

Nitric oxide (NO) is a gaseous signaling molecule with known prominent roles in plant development and abiotic stress acclimation. This study reports that NO plus nitrogen (N) conspicuously alleviated cadmium (Cd)-inhibited photosynthetic activity by interacting with ethylene synthesis, and modifying the mechanisms of antioxidant enzymes, N and sulfur (S) assimilation in mustard (Brassica juncea L.) cv. Giriraj plants. The application of 100 µM sodium nitroprusside (SNP; NO donor) and 200 mg N kg−1 soil (as ammonium nitrate) to 200 mg Cd kg−1 soil-grown plants reduced oxidative stress (superoxide radical, hydrogen peroxide, and thiobarbituric acid reactive species content) and cell death through enhanced synthesis of metal chelators (non-protein thiols and phytochelatins) and limited Cd translocation. The reduced cellular ROS was also related to NO and N-mediated enhancement of S and N-assimilation, and antioxidants. The NO and N-mediated higher expression of psbA and psbB genes of D1 protein of PSII resulted into higher photosynthetic performance even under higher Cd regimes. Further, NO and N in the presence of aminoethoxyvinylglycine (AVG, ethylene biosynthesis inhibitor) under Cd stress showed reduced growth and photosynthetic attributes, with greater oxidative stress, substantiating ethylene involvement in NO and N-mediated reversal of Cd-inhibited photosynthesis. The principal component analysis showed a positive correlation between the treatment SNP + N + Cd and NO production, antioxidant enzymes, glutathione and metal chelators which resulted in higher N and S-assimilation, and better growth and photosynthetic capacity under Cd stress. The results indicate that NO-mediated alleviation of Cd toxicity and protection of photosynthetic activity were more conspicuous with N availability through the coordination with ethylene synthesis. The outcome of the study may be utilized in maintaining and managing hostile soils polluted with potentially toxic metals like Cd.

Sunlight-induced repair of photosystem II in moss Semibarbula orientalis under submergence stress.

Lower plants such as bryophytes often encounter submergence stress, even in low precipitation conditions. Our study aimed to understand the mechanism of submergence tolerance to withstand this frequent stress in moss (Semibarbula orientalis) during the day and at night. These findings emphasise that light plays a crucial role in photoreactivation of PSII in S. orientalis, which indicates that light not only fuels photosynthesis but also aids in repairing the photosynthetic machinery in plants. Submergence negatively affects photosynthesis parameters such as specific and phenomenological fluxes, density of functional PSII reaction centres (RC/CS), photochemical and non-photochemical quenching (Kp and Kn), quantum yields (ϕP0, ϕE0, ϕD0), primary and secondary photochemistry, performance indices (PIcs and PIabs), etc. Excessive antenna size caused photoinhibition at the PSII acceptor side, reducing the plastoquinone pool through the formation of PSII triplets and reactive oxygen species (ROS). This ROS-induced protein and PSII damage triggered the initiation of the repair cycle in presence of sunlight, eventually leading to the resumption of PSII activity. However, ROS production was regulated by antioxidants like superoxide dismutase (SOD) and catalase (CAT) activity. The rapid recovery of RS/CS observed specifically under sunlight conditions emphasises the vital role of light in enabling the assembly of essential units, such as the D1 protein of PSII, during stress in S. orientalis. Overall, light is instrumental in restoring the photosynthetic potential in S. orientalis growing under submergence stress. Additionally, it was observed that plants subjected to submergence stress during daylight hours rapidly recover their photosynthetic performance. However, submergence stress during the night requires a comparatively longer period for the restoration of photosynthesis in the moss S. orientalis.

Time-course effects of Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) on Chlorella pyrenoidosa: Growth inhibition and adaptability mechanisms

Tris(1,3-dichloro-2-propyl) phosphate (TDCPP), a widely used chlorinated organophosphorus flame retardant, is an increasingly widespread contaminant of aquatic environment. In this study, time-dependent effect of TDCPP on the freshwater green-algae Chlorella pyrenoidosa was investigated and its underlying mechanisms were explored. We show that TDCPP lower than 10 ppm caused a reversible inhibition of algal growth, with complete inhibition occurring at 15 ppm. This inhibition was not caused by damage from reactive oxygen species, but rather resulted from the impairment of photosynthetic function, with PSII reaction center as the primary target, as indicated by Chl a fluorescence induction, QA− reoxidation, S-state distribution and immunoblot analysis. The reversal of damage caused by TDCPP concentrations under 10 ppm might be attributable to the repair of photosynthetic function by de novo protein biosynthesis in the chloroplast, with the most likely explanation being the replacement of the damaged PSII D1 protein. The results provide novel insights into mechanisms of TDCPP toxicity toward freshwater microalgae and better understanding of ecological consequences of TDCPP in the environment.

Ferredoxin5 Deletion Affects Metabolism of Algae during the Different Phases of Sulfur Deprivation.

Ferredoxin5 (FDX5), a minor ferredoxin protein in the alga Chlamydomonas (Chlamydomonas reinhardtii), helps maintain thylakoid membrane integrity in the dark. Sulfur (S) deprivation has been used to achieve prolonged hydrogen production in green algae. Here, we propose that FDX5 is involved in algal responses to S-deprivation as well as to the dark. Specifically, we tested the role of FDX5 in both the initial aerobic and subsequent anaerobic phases of S-deprivation. Under S-deprived conditions, absence of FDX5 causes a distinct delay in achieving anoxia by affecting photosynthetic O2 evolution, accompanied by reduced acetate uptake, lower starch accumulation, and delayed/lower fermentative metabolite production, including photohydrogen. We attribute these differences to transcriptional and/or posttranslational regulation of acetyl-CoA synthetase and ADP-Glc pyrophosphorylase, and increased stability of the PSII D1 protein. Interestingly, increased levels of FDX2 and FDX1 were observed in the mutant under oxic, S-replete conditions, strengthening our previously proposed hypothesis that other ferredoxins compensate in response to a lack of FDX5. Taken together, the results of our omics and pull-down experiments confirmed biochemical and physiological results, suggesting that FDX5 may have other effects on Chlamydomonas metabolism through its interaction with multiple redox partners.

Unstandardized UV-C dose used for killing harmful cyanobacteria may instead initiate accelerated growth in the target organisms

Although UV-C radiation has been in use for killing unwanted cyanobacteria, experiments with lower doses of UV-C radiation instead showed induction of growth related parameters and enhanced biomass production in the cyanobacterium Nostoc muscorum Meg1. When the cyanobacterial cultures were exposed to UV-C radiation of varying doses (6, 12 and 18 mJ/cm2), concentrations of various photo-absorbing pigments, RuBisCO and D1 protein of PSII; activities of oxygen evolving complex, nitrogenase and glutamine synthetase were significantly increased upon 6 and 12 mJ/cm2 UV-C radiation exposures. Resulting higher photosynthetic performance was evident from the augmentation in carbohydrate content by ∼49% under single exposure to 6 mJ/cm2 UV-C by fifteenth day. The increased performances of both RuBisCO and D1 proteins were in part also due to induction at the genetic level as seen from the increase in their mRNA and protein levels under treatment. Similar increase was also observed in protein (16%) and in lipid contents (43%) that reflected an upsurge in the total biomass. Highest biomass (463 mg/L/d) was noted in culture exposed to 6 mJ/cm2 UV-C radiation, representing a ∼25% increase. Furthermore the possibility of this organism using part of the incident UV-C radiation as an additional source of energy was deduced from an experiment where the thylakoid membranes excited within UV (226–400 nm) range showed emission at longer wavelengths with an emission maximum at ∼640 nm. Thus this work provides evidence that lower UV-C doses can potentially augment cyanobacterial growth and use of unstandardized UV-C doses for restricting cyanobacterial growth may in fact produce contrary result.

Dose dependent variance in UV-C radiation induced effects on carbon and nitrogen metabolism in the cyanobacterium Nostoc muscorum Meg1

With the intention of getting an insight into the differential effect of UV-C radiation on the N2-fixing heterocystous cyanobacterium Nostoc muscorum Meg1, various aspects of carbon and nitrogen metabolism was evaluated in the organism. Exposure to different doses of UV-C (6, 12, 18 and 24 mJ/cm2) showed that among various photo-absorbing pigments, phycobiliproteins were most sensitive. Oxygen evolving complex (OEC) activity measured as net oxygen evolution rate decreased by 63% upon 24 mJ/cm2 exposure. Western blot analysis established that D1 protein of PSII was highly sensitive and its levels decreased even at a radiation dose as low as 6 mJ/cm2. In contrast, levels of the Calvin cycle enzyme RuBisCO was increased at 6 and 12 mJ/cm2 doses but the level decreased drastically (84%) at higher dose (24 mJ/cm2). The nitrogenase enzyme activity decreased at all doses but the ammonia assimilating enzyme glutamine synthetase (GS) activity recorded increase at the lower doses. The reactive oxygen species (ROS) and lipid peroxidation increased upon UV-C exposure. Transmission electron microscopic observation revealed damage to ultrastructure especially the thylakoid membrane organization, aggregation of dissolving phycobilisomes and loss of caboxysomes. Interestingly, sub-lethal radiation (6 and 12 mJ/cm2) dose exposures increased the growth rate in the organism when growth was measured over a period of 11 days after radiation exposure.

Dysregulation of photosynthetic genes in oceanic Prochlorococcus populations exposed to organic pollutants

The impact of organic pollutants on oceanic ecosystem functioning is largely unknown. Prochlorococcus, the most abundant known photosynthetic organism on Earth, has been suggested to be especially sensible to exposure to organic pollutants, but the sub-lethal effects of organic pollutants on its photosynthetic function at environmentally relevant concentrations and mixtures remain unexplored. Here we show the modulation of the expression of two photosynthetic genes, rbcL (RuBisCO large subunit) and psbA (PSII D1 protein), of oceanic populations of Prochlorococcus from the Atlantic, Indian and Pacific Oceans when exposed to mixtures of organic pollutants consisting of the non-polar fraction of a seawater extract. This mixture included most persistent organic pollutants, semivolatile aromatic-like compounds, and the unresolved complex mixture of hydrocarbons. Prochlorococcus populations in the controls showed the expected diel cycle variations in expression of photosynthetic genes. However, exposure to a complex mixture at concentrations only 2-fold above the environmental levels resulted in a decrease of expression of both genes, suggesting an effect on the photosynthetic function. While organic pollutant effects on marine phytoplankton have been already demonstrated at the cellular level, this is the first field study showing alterations at the molecular level of the photosynthetic function due to organic pollutants.

Quality Control of PSII: Behavior of PSII in the Highly Crowded Grana Thylakoids Under Excessive Light

The grana thylakoids of higher plant chloroplasts are crowded with PSII and the associated light-harvesting complexes (LHCIIs). They constitute supercomplexes, and often form semi-crystalline arrays in the grana. The crowded condition of the grana may be necessary for efficient trapping of excitation energy by LHCII under weak light, but it might hinder proper movement of LHCII necessary for reversible aggregation of LHCII in the energy-dependent quenching of Chl fluorescence under moderate high light. When the thylakoids are illuminated with extreme high light, the reaction center-binding D1 protein of PSII is photodamaged, and the damaged protein migrates to the grana margins for degradation and subsequent repair. In both moderate and extreme high-light conditions, fluidity of the thylakoid membrane is crucial. In this review, we first provide an overview of photoprotective processes, then discuss changes in membrane fluidity and mobility of the protein complexes in the grana under excessive light, which are closely associated with photoprotection of PSII. We hypothesize that reversible aggregation of LHCII, which is necessary to avoid light stress under moderate high light, and swift turnover of the photodamaged D1 protein under extreme high light are threatened by irreversible protein aggregation induced by reactive oxygen species in photochemical reactions.

The UV-B photoreceptor UVR8 promotes photosynthetic efficiency in Arabidopsis thaliana exposed to elevated levels of UV-B

The UV-B photoreceptor UVR8 regulates expression of genes in response to UV-B, some encoding chloroplast proteins, but the importance of UVR8 in maintaining photosynthetic competence is unknown. The maximum quantum yield of PSII (F (v)/F(m)) and the operating efficiency of PSII (Φ(PSII)) were measured in wild-type and uvr8 mutant Arabidopsis thaliana. The importance of specific UVR8-regulated genes in maintaining photosynthetic competence was examined using mutants. Both F (v)/F(m) and Φ(PSII) decreased when plants were exposed to elevated UV-B, in general more so in uvr8 mutant plants than wild-type. UV-B increased the level of psbD-BLRP (blue light responsive promoter) transcripts, encoding the PSII D2 protein. This increase was mediated by the UVR8-regulated chloroplast RNA polymerase sigma factor SIG5, but SIG5 was not required to maintain photosynthetic efficiency at elevated UV-B. Levels of the D1 protein of PSII decreased markedly when plants were exposed to elevated UV-B, but there was no significant difference between wild-type and uvr8 under conditions where the mutant showed increased photoinhibition. The results show that UVR8 promotes photosynthetic efficiency at elevated levels of UV-B. Loss of the DI polypeptide is probably important in causing photoinhibition, but does not entirely explain the reduced photosynthetic efficiency of the uvr8 mutant compared to wild-type.

Dependence of reaction center-type energy-dependent quenching on photosystem II antenna size

The effects of photosystem II antenna size on reaction center-type energy-dependent quenching (qE) were examined in rice plants grown under two different light intensities using both wild type and qE-less (OsPsbS knockout) mutant plants. Reaction center-type qE was detected by measuring non-photochemical quenching at 50 μmol photons m − 2 s − 1 white light intensity. We observed that in low light-grown rice plants, reaction center-type qE was higher than in high light-grown plants, and the amount of reaction center-type qE did not depend on zeaxanthin accumulation. This was confirmed in Arabidopsis npq1–2 mutant plants that lack zeaxanthin due to a mutation in the violaxanthin de-epoxidase enzyme. Although the electron transport rate measured at a light intensity of 50 μmol photons m − 2 s − 1 was the same in high light- and low light-grown wild type and mutant plants lacking PsbS protein, the generation of energy-dependent quenching was completely impaired only in mutant plants. Analyses of the pigment content, Lhcb proteins and D1 protein of PSII showed that the antenna size was larger in low light-grown plants, and this correlated with the amount of reaction center-type qE. Our results mark the first time that the reaction center-type qE has been shown to depend on photosystem II antenna size and, although it depends on the existence of PsbS protein, the extent of reaction center-type qE does not correlate with the transcript levels of PsbS protein. The presence of reaction center-type energy-dependent quenching, in addition to antenna-type quenching, in higher plants for dissipation of excess light energy demonstrates the complexity and flexibility of the photosynthetic apparatus of higher plants to respond to different environmental conditions.

광 스트레스에 대한 고추 잎의 광합성 반응과 광 보호 메커니즘

Photoinhibition and photoprotection of PSII in the leaves of hot pepper (Capsicum annuum L.) grown in Hoagland solution and Tap water were compared. Though changes in the rates of <TEX>$O_2$</TEX> evolution as a function of photon fluence rate (PFR) were comparable, the rates of respiration in the dark was 3 times higher in the Hoagland solution grown leaves than in the Tap-water grown ones. Compared to Hoagland solution grown plane, PSIIs of Tap water grown pepper leaves were more susceptible to photoinhibitory light treatment. In order to inactivate functional PSII to the same extents, Hoagland solution grown plants required almost 2-fold high light <TEX>$(1600{\mu}molm^{-2}s^-)$</TEX> treatment than those of Tap water <TEX>$(900{\mu}molm^{-2}s^-)$</TEX>. Interestingly, the remaining fraction of PSII in Hoagland grown pepper was able to survive under prolonged illumination in the presence of lincomycin, which probably means that the growth condition of plant seemed to have an effect on the recovery of PSII from light stress. When PSII was severly photoinactivated at a chilling temperature, recovery was observed only if the residual functional PSII were not inhibited with DCMU, Nigericin and MV during recovery. In conclusion, PSIIs grown in the Hoagland solution was more resistant to excess light than in the Tap water grown one and the recovery of PSII from photodamage was more efficient in Hoagland grown pepper leaves than Tap water grown one, which means that the increased dark respiration may play a important role in the protection of PSII from photoinhibition by helping repair photosynthetic proteins (in particular, the D1 protein of PSII) degraded by photoinhibition.

Effects of long-term ionic and osmotic stress conditions on photosynthesis in the cyanobacterium Synechocystis sp. PCC 6803.

Salinity is considered to be one of the most severe problems in worldwide agricultural production, but the published investigations give contradictory results of the effect of ionic and osmotic stresses on photosynthesis. In the present study, long-term effects of both ionic and osmotic stresses, especially on photosynthesis, were investigated using the moderately halotolerant cyanobacterium Synechocystis sp. PCC 6803. Our results show that the PSII activity and the photosynthetic capacity tolerated NaCl but a high concentration of sorbitol completely inhibited both activities. In line with these results, we show that the amount of the D1 protein of PSII was decreased under severe osmotic stress, whereas the levels of PsaA / B and NdhF3 proteins remained unchanged. However, high concentrations of sorbitol stress led to a drastic decrease of both psbA (encoding D1) and psaA (encoding PsaA) transcripts, suggesting that severe osmotic stress may abolish the tight coordination of transcription and translation normally present in bacteria, at least in the case of the psaA gene. Taken together, our results indicate that the osmotic stress component is more detrimental to photosynthesis than the ionic one and, furthermore, under osmotic stress, the D1 protein appears to be the target of this stress treatment.

Characterization of senescence-associated gene expression and senescence-dependent and -independent cysteine proteases differing in microsomal processing in Anthurium

This study was conducted to identify senescence-regulated signaling pathways and genes in the monocot Anthurium andraeanum L. for future efforts to modify leaf senescence via biotechnology. The Arabidopsis senescence-activated promoter (Pr- SAG12) fused to the gfp4 gene was expressed 20-fold higher, while the Pr35S- gfp4 was expressed 30% lower, in senescing relative to healthy leaves of Anthurium. This indicated that a senescence-activated pathway correctly regulated the Arabidopsis sag12 cysteine protease gene and suggested the presence of sag12 homologues in Anthurium. To identify sag12 homologs, two different Anthurium cysteine protease cDNAs, anth16 and anth17, were cloned and their sequences, expression, and protein products were characterized. The Anth17 and Anth16 protein sequences were 58 and 51% identical with Sag12, respectively. Of the two genes, anth17 expression most closely resembled the sag12 gene. Anth16 was expressed primarily in healthy immature leaves, while anth17 and a control gene (ubiquitin) were significantly induced during senescence of mature leaves. The levels of known internal standards, chlorophyll, mRNAs for cab (chlorophyll-a,b-binding protein) and psbA (D1 protein of PSII), decreased during senescence and served to verify the progression of senescence. Cytokinin treatments repressed anth17 mRNA accumulation and sustained psbA mRNA levels in senescing leaves. Sucrose moderately reduced anth17 and psbA mRNA levels. Anth16 has a unique secretory vesicle sorting motif in the C-terminus, and produces a 70 kDa in vitro translation product that is post-translationally processed to 57 kDa by microsomal membranes, a property of secretory proteins. In contrast, anth17 was not processed and its intracellular retention is proposed to be necessary for senescence-dependent cellular disassembly and proteolysis.

ASTAXANTHIN ACCUMULATION IN HAEMATOCOCCUS PLUVIALIS (CHLOROPHYCEAE) AS AN ACTIVE PHOTOPROTECTIVE PROCESS UNDER HIGH IRRADIANCE1

Green cells of Haematococcus pluvialis Flotow accumulate the ketocarotenoid astaxanthin under stress conditions, such as high irradiance, nutrient deficiency, high salinity, and high temperature. Though some photoprotective mechanisms have been suggested, the function of astaxanthin in red cysts is still questioned. We studied the role of astaxanthin in photoprotection by inducing its formation in logarithmically growing cultures by high irradiance, thus avoiding unrelated processes that can occur in H. pluvialis when carotenogenesis is induced by other stresses. On exposure to high irradiance, the green Haematococcus culture turned red as lipid globules loaded with astaxanthin esters were formed and concentrated at the periphery of the cell. During this phase of induction, the photosynthesis rates remained high, but the amount of the D1 protein of PSII was significantly reduced. The decline in D1 protein content stopped after 1 day; the level then increased, returning to normal after 5 days. The response of the D1 protein was indicative of a transitional phase in the acclimation of Haematococcus to high light. The formation and deposition of astaxanthin seemed to prevent further reduction in D1 protein level, thus enabling the cell to maintain PSII function and structural integrity. This result seems to be a clear indication of the light screening by astaxanthin, which absorbs light in the blue region, thus protecting the photosynthetic apparatus. When the cells recovered from the high light stress, the astaxanthin globules concentrated around the nucleus, indicating that the pigment also serves as a physicochemical barrier, protecting the replicating DNA from oxidation as the cells divide.

Throwing light on photosynthesis

Molecular mechanisms of photosynthesisby Robert E. Blankenship, Blackwell Science, 2002. US$49.95/£29.95 (352 pages) ISBN 0 632 04321 0It has long been known that light can be converted into useful chemical energy. The importance of this photosynthetic process has been recognized in different ways throughout history, and was elegantly acknowledged by Julius Robert Mayer in 1845 when he wrote: ‘Nature set herself the task of capturing the light flooding towards the earth and storing this, the most elusive of all forces, by converting it into an immobile force…the plant world constitutes a reservoir in which the fleeting sun rays are fixed and ingeniously stored for future use, a providential measure to which the very existence of the human race is inescapably bound’.In his recently published book, Bob Blankenship replaces Mayer's flowery definition of photosynthesis with ‘a process in which light energy is captured and stored by an organism and the stored energy is used to drive cellular processes’. Behind this simple and rather bland modern definition of photosynthesis lies a vast base of knowledge, encompassing disciplines ranging from photophysics and photochemistry to molecular biology and physiology. The reactions of photosynthesis are initiated by the absorption of quanta by pigments, which occurs in the femtosecond time domain, followed by reactions that proceed in a time-scale stretching to days or even years. Therefore, to write a comprehensive text book on the molecular mechanisms of photosynthesis is a daunting task. Blankenship has taken up this challenge and, in my view, has produced a first class product. His approach is sensitive to the student reader and, with this in mind, he has included a 47-page appendix in which the appropriate basic principles of photophysics are well described. The remaining 257 pages comprise 11 chapters, in which Blakenship leads the reader through a logical progression of photosynthesis. The first three chapters set the scene, giving basic concepts in terms of reactions, where they take place and how they were discovered. This foundation is then built upon with a thorough coverage of structural and spectral properties of the photosynthetic pigments, and of the means by which they are associated with proteins. Over the past few years, X-ray crystallography has provided spectacular insights into how these pigment proteins are arranged, and has also given a structural basis to experimental and theoretical considerations of the energy transfer mechanisms. All this is dealt with rigorously by Blankenship. His account of reaction centre complexes in Chapter 6 deserves equal praise. He could not be more up-to-date in his content, providing the reader, for example, with a discussion of the latest X-ray structures of photosystems (PS) I and II. Chapter 7 is a bit of a hodgepodge. Quite rightly, Blankenship moves on to other components of the photosynthetic electron transport chain. He gives details of the similarities of the bacterial cytochrome bc and chloroplast cytochrome b6f complexes, and discusses the role of diffusable redox cofactors such as plastoquinone, plastocyanin, ferredoxin and NADPH. However, the discussion of turnover of the D1 protein of PSII and chlorophyll fluorescence as a probe of PSII function seems, to me, to be misplaced in this chapter.The conversion of ADP to ATP, the role of proton gradients, and Mitchell's chemiosmotic hypothesis are detailed in a useful and readable way in Chapter 8. In addition, this chapter focuses on speculations about the rotatory ATP synthase mechanism by drawing on the most recent structural information available.It has been traditional to consider the processes of photosynthesis as consisting of two stages: light and dark reactions. Blankenship has continued this tradition and uses Chapter 9 to discuss biochemical details of the dark reactions; namely carbon fixation via C3, C4 and CAM metabolism. He also includes the CO2 concentrating mechanism found in aquatic photosynthetic systems, and the biochemical reactions involved in long-term storage of photosynthate as sucrose and starch.The photosynthetic apparatus, whether it be of anaerobic bacteria or aerobic cyanobacteria, algae and higher plants, is complicated, being composed of many hundreds of different proteins, enzymes and cofactors. How these complex systems assemble themselves and how they evolved are the subjects of the last two chapters, respectively. Blankenship has a keen interest in the evolution of photosynthesis and his chapter on this subject makes good reading. Although there are major gaps in our knowledge, recent advances in gene sequencing and structural analyses of photosynthetic proteins are providing fodder for fascinating and realistic hypotheses; these provide a grand finale for this excellent text book. I will have no hesitation in recommending it to my students and colleagues alike.

Rapid recovery of photosystems on rewetting desiccation-tolerant mosses: chlorophyll fluorescence and inhibitor experiments.

In the mosses Racomitrium lanuginosum, Anomodon viticulosus and Rhytidiadelphus loreus, after a few days air dry, F:(v)/F:(m) reached, within the first minute of remoistening in the dark, two-thirds or more of the value attained after 40 min. A fast initial phase of recovery was completed within 10-20 min after which further change was slow. Initial recovery of Phi(PSII) in the light was somewhat slower, but was generally substantially complete within a similar time. Remoistening with 0.3 mM cycloheximide (CHX) or 3 mM dithiothreitol (DTT) made little difference to this short-term (40 min) recovery of either F:(v)/F:(m) or Phi(PSII); 3 mM chloramphenicol (CMP) had little effect on recovery of F:(v)/F:(m), but resulted in substantial (though not total) depression of Phi(PSII) and (14)CO(2) uptake. Effects of the protein-synthesis inhibitors and DTT were much more clearly apparent in longer-term experiments (>20 h) but only in the light. In the dark, the three inhibitors had at most only slight effects over periods of 60-100 h. In the light, CMP-treated samples of all three species showed a progressive decline of dark-adapted F:(v)/F:(m), falling to zero within 1-5 d (possibly due to blocking of the turnover of the D1 protein of PSII) and accelerated by DTT. CHX-treated samples showed a similar but slower decline. In the shade-adapted and relatively desiccation-sensitive Rhytidiadelphus loreus, slow recovery of F:(v)/F:(m) continued in the dark even in the presence of CMP and CHX for much of the 142 h of the experiment. The results indicate that in desiccation-tolerant bryophytes recovery of photosynthesis after periods of a few days air dry requires only limited chloroplast protein synthesis and is substantially independent of protein synthesis in the cytoplasm.

Chloroplast proteases: Possible regulators of gene expression?

A wide range of proteolytic processes in the chloroplast are well recognized. These include processing of precursor proteins, removal of oxidatively damaged proteins, degradation of proteins missing their prosthetic groups or their partner subunit in a protein complex, and adjustment of the quantity of certain chloroplast proteins in response to changing environmental conditions. To date, several chloroplast proteases have been identified and cloned. The chloroplast processing enzyme is responsible for removing the transit peptides of newly imported proteins. The thylakoid processing peptidase removes the thylakoid-transfer domain from proteins translocated into the thylakoid lumen. Within the lumen, Tsp removes the carboxy-terminal tail of the precursor of the PSII D1 protein. In contrast to these processing peptidases which perform a single endo-proteolytic cut, processive proteases that can completely degrade substrate proteins also exist in chloroplasts. The serine ATP-dependent Clp protease, composed of the proteolytic subunit ClpP and the regulatory subunit ClpC, is located in the stroma, and is involved in the degradation of abnormal soluble and membrane-bound proteins. The ATP-dependent metalloprotease FtsH is bound to the thylakoid membrane, facing the stroma. It degrades unassembled proteins and is involved in the degradation of the D1 protein of PSII following photoinhibition. DegP is a serine protease bound to the lumenal side of the thylakoid membrane that might be involved in the chloroplast response to heat. All these peptidases and proteases are homologues of known bacterial enzymes. Since ATP-dependent bacterial proteases and their mitochondrial homologues are also involved in the regulation of gene expression, via their determining the levels of key regulatory proteins, chloroplast proteases are expected to play a similar role.

Put out the light, and then put out the light

The lowest photon flux density of photosynthetically active radiation at which O2-evolving marine photolithotrophs appear to be able to grow is some 10 nmol photon m−2 s−1, while marine non-O2-evolvers can grow at 4 nmol photon m−2 s−1, in both cases with the photon flux density averaged over the 24 hour L:D cycle. Constraints on the ability to grow at very low fluxes of photosynthetically active radiation fall into three categories. Category one includes essential processes whose efficiency is independent of the rate of energy input, but whose catalysts show phylogenetic variation leading to different energy costs for a given process in different taxa, e.g. light-harvesting complexes, RUBISCO and probably in the sensitivity of PsII to photodamage. The second category comprises essential processes whose efficiency decreases with decreasing energy input rate as a result of back-reactions independent of the energy input rate, e.g. charge recombination following charge separation by PsII and short-circuit H+ fluxes across the thylakoid membrane which decrease the fraction of pumped H+ which can be used in adenosine diphosphate phosphorylation. Category two also includes that component of protein turnover which cannot be related to replacement of polypeptides which were incorrectly assembled following uncorrected errors of transcription or translation, or which were damaged by processes whose rate increases with increasing energy input rate such as photodamage to PsII. The third category includes only O2-dependent damage to the D1 protein of PsII whose rate increases with a decreasing incident flux of photosynthetically active radiation. Processes in categories two and three are most likely to impose the lower limit on the photon flux density which can support photolithotrophic growth. The available literature, mainly on organisms which are not adapted to growth at very low photon flux densities, suggests that three major limitations (charge recombination in PsII, H+ leakage and slippage, and protein turnover) can individually impose lower limits in excess of 20 nmol photon m−2 s−1 on photolithotrophic growth. Furthermore, these three limitations are interactive, so that considering all three processes acting in series leads to an even higher predicted lower photon flux density limit for photolithotrophic growth.