Three-state mathematical model for the assessment of DCMU-treated photosystem II heterogeneity.

Photosystem II (PSII) is one of the main pigment-protein complexes of photosynthesis which is highly sensitive to unfavorable environmental factors. The heterogeneity of PSII properties is essential for the resistance of autotrophic organisms to stress factors. Assessment of the PSII heterogeneity may be used in environmental monitoring for on-line detection of contamination of the environment. We propose an approach to assess PSII oxygen-evolving complex and light-harvesting antenna heterogeneity that is based on mathematical modeling of the shape of chlorophyll a fluorescence rise of 3-(3,4-dichlorophenyl)-1,1-dimethylurea-treated samples. The hierarchy of characteristic times of the processes considered in the model makes it possible to reduce the model to a system of three ordinary differential equations. The analytic solution of the reduced three-state model is expressed as a sum of two exponential functions, and it exactly reproduces the solution of the complete system within the time range from microseconds to hundreds of milliseconds. The combination of several such models for reaction centers with different properties made it possible to use it as an instrument to study PSII heterogeneity. PSII heterogeneity was studied for Chlamydomonas at different intensities of actinic light, for Scenedesmus under short-term heating, and for Chlorella grown in nitrate-enriched and nitrate-depleted media.

Nanodiamond Particles Reduce Oxidative Stress Induced by Methyl Viologen and High Light in the Green Alga Chlamydomonas reinhardtii.

Widely used in biomedical and bioanalytical applications, the detonation nanodiamonds (NDs) are generally considered to be biocompatible and non-toxic to a wide range of eukaryotic cells. Due to their high susceptibility to chemical modifications, surface functionalisation is often used to tune the biocompatibility and antioxidant activity of the NDs. The response of photosynthetic microorganisms to redox-active NDs is still poorly understood and is the focus of the present study. The green microalga Chlamydomonas reinhardtii was used to assess the potential phytotoxicity and antioxidant activity of NDs hosting hydroxyl functional groups at concentrations of 5-80 µg NDs/mL. The photosynthetic capacity of microalgae was assessed by measuring the maximum quantum yield of PSII photochemistry and the light-saturated oxygen evolution rate, while oxidative stress was assessed by lipid peroxidation and ferric-reducing antioxidant capacity. We demonstrated that hydroxylated NDs might reduce cellular levels of oxidative stress, protect PSII photochemistry and facilitate the PSII repair under methyl viologen and high light associated stress conditions. Factors involved in this protection may include the low phytotoxicity of hydroxylated NDs in microalgae and their ability to accumulate in cells and scavenge reactive oxygen species. Our findings could pave the way for using hydroxylated NDs as antioxidants to improve cellular stability in algae-based biotechnological applications or semi-artificial photosynthetic systems.

Acclimation Response of Green Microalgae Chlorella Sorokiniana to 2,3',4,4',6-Pentachlorobiphenyl.

The effect of the toxicant 2,3',4,4',6-pentachlorobiphenyl (PCB-119) on the growth, chlorophyll content, and PSII activity of C. sorokiniana cells was investigated. A strong negative effect of the toxicant was observed at PCB concentration of 0.05 µg mL-1 , when culture growth ceased, chlorophyll strongly bleached, and cell death occurred. The use of original highly sensitive fluorimeter to measure three types of high-resolution chlorophyll fluorescence kinetics allowed us to detect an initial dramatic decrease in the activity of primary photosynthetic reactions, followed by their almost complete recovery at the end of the incubation period when most cells were dead. The study of the distribution of individual cells in culture in terms of Fv /Fm parameter, which reflects the quantum yield of PSII photochemistry, revealed the existence of 2-3% of cells retaining high Fv /Fm (>0.7) in the presence of the toxicant. The treated cultures were able to resume growth after prolonged incubation in fresh medium. The high sensitivity fluorescence methods used made it possible to identify stress-resistant cells which maintain high photosynthetic activity in the presence of lethal doses of toxic substances; these cells provide recovery of the population after stress.

Single-walled carbon nanotubes protect photosynthetic reactions in Chlamydomonas reinhardtii against photoinhibition.

Single-walled carbon nanotubes (SWCNTs) are among the most exploited carbon allotropes in nanosensing, bioengineering, and photobiological applications, however, the interactions of nanotubes with the photosynthetic process and structures are still poorly understood. We found that SWCNTs are not toxic to the photosynthetic apparatus of the model unicellular alga Chlamydomonas reinhardtii and demonstrate that this carbon nanomaterial can protect algal photosynthesis against photoinhibition. The results show that the inherent phytotoxicity of the nanotubes may be overcome by an intentional selection of nanomaterial characteristics. A low concentration (2 µg mL-1) of well-dispersed, purified and small SWCNTs did not alter the growth and pigment accumulation of the cultures. Indeed, under the photoinhibitory conditions of our experiments, SWCNT-enriched samples were characterized by a lower rate of PSII inactivation, reduced excitation pressure in PSII, a higher rate of photosynthetic electron transport, and an increased non-photochemical quenching in comparison with the controls. In addition, SWCNTs change the distribution of energy between the photosystems in favour of PSII (state 1). The underlying mechanism of this action is not yet understood but possibly, electrons or energy can be exchanged between the redox active nanotubes and photosynthetic components, and probably other redox active intra-chloroplast constituents. Alternatively, nanotubes may promote the formation of an NPQ conformation of PSII. Our results provided evidence for such electron/energy transfer from photosynthetic structures toward the nanotubes. The discovered photoprotective effects can potentially be used in photobiotechnology to maintain the photosynthetic activity of microorganisms under unfavourable conditions.

Multiple in vivo Effects of Cadmium on Photosynthetic Electron Transport in Pea Plants.

The inhibitory effects of cadmium (CdSO4 ) on the primary photosynthetic processes were studied in vivo in Pisum sativum by using Multi-function Plant Efficiency Analyser (M-PEA-2). Photosynthetic parameters related to photosystem (PS) II, PS I and intersystem electron carriers were calculated from the light-induced kinetics of prompt chlorophyll a fluorescence (OJIP transient), delayed chlorophyll a fluorescence (DF), and 820 nm modulated reflection (MR). Low-dose exposure to cadmium (20 µm CdSO4 for 48 h) reduced probability of electron transfer from plastoquinones to the terminal electron acceptors of PSI (δRo ) accompanied by a decrease in the rate of P700 + and PC reduction (Vred ) and the magnitude of the I2 step on the DF kinetics. Electron transport through PSI remained unaltered. The obtained results allowed us to propose existence of the potential site of inhibition of photosynthetic electron flow by cadmium between PSII and PSI. We propose to use parameters δRo , Vred , and I2 /I1 as sensitive indicators of an early contamination by heavy metals.

Immobilized heterocysts as microbial factories for sustainable nitrogen fixation.

A novel thin-layer biocatalyst for photosynthetic N2 fixation and H2 photoproduction was assembled using a Ca2+-alginate matrix and heterocysts isolated from wild-type Anabaena sp. PCC 7120 filaments. Compared to suspension heterocysts, heterocysts entrapped in Ca2+-alginate films showed improved stability of the nitrogenase system. While suspension heterocysts lost nitrogenase activity within 24 h, immobilized heterocysts supported nitrogenase activity for up to 125 h. The maximum specific rate of acetylene reduction was the same in both cases (∼0.4 µmol C2H2 mg Chl-1 h-1), but the catalyst with entrapped heterocysts required a much longer time to achieve the maximum rate (60 h instead of 3 h in suspension). Simultaneously with acetylene reduction, the immobilized heterocysts were able to photoproduce H2 for 125 h, yielding up to 1.1 mmol H2 mg Chl-1. The absence of acetylene increased the H2 photoproduction rate to a maximum of 25-30 µmol H2 mg Chl-1 h-1, and the catalyst was capable of H2 photoproduction for 190 h, yielding up to 2.5 mmol H2 mg Chl-1. The recovery of the catalyst with entrapped heterocysts was achieved through placing the cells in a N2 atmosphere for 24 h. This engaged a second cycle of H2 photoproduction, which lasted for another 240 h (10 days), thus yielding ∼3 mmol H2 mg Chl-1 in total after 454 h. Together, these findings demonstrate great potential for a heterocyst-based thin-layer platform for the sustainable production of chemicals and biofuels.

Chlorophyll fluorescence induction and relaxation system for the continuous monitoring of photosynthetic capacity in photobioreactors.

The development of high-performance photobioreactors equipped with automatic systems for non-invasive real-time monitoring of cultivation conditions and photosynthetic parameters is a challenge in algae biotechnology. Therefore, we developed a chlorophyll (Chl) fluorescence measuring system for the online recording of the light-induced fluorescence rise and the dark relaxation of the flash-induced fluorescence yield (Qa- - re-oxidation kinetics) in photobioreactors. This system provides automatic measurements in a broad range of Chl concentrations at high frequency of gas-tight sampling, and advanced data analysis. The performance of this new technique was tested on the green microalgae Chlamydomonas reinhardtii subjected to a sulfur deficiency stress and to long-term dark anaerobic conditions. More than thousand fluorescence kinetic curves were recorded and analyzed during aerobic and anaerobic stages of incubation. Lifetime and amplitude values of kinetic components were determined, and their dynamics plotted on heatmaps. Out of these data, stress-sensitive kinetic parameters were specified. This implemented apparatus can therefore be useful for the continuous real-time monitoring of algal photosynthesis in photobioreactors.

Comparative analyses of H2 photoproduction in magnesium- and sulfur-starved Chlamydomonas reinhardtii cultures.

Magnesium (Mg)-deprived Chlamydomonas reinhardtii cells are capable to sustain hydrogen (H2 ) photoproduction at relatively high photosystem II (PSII) activity levels for an extended time period as compared with sulfur (S)-deprived cells. Herein, we present a comparative study of H2 photoproduction induced by Mg and S shortage to unravel the specific rearrangements of the photosynthetic machinery and cell metabolism occurring under the two deprivation protocols. The exhaustive analysis of photosynthetic activity and regulatory pathways, respiration and starch metabolism revealed the specific rearrangements of the photosynthetic machinery and cellular metabolism, which occur under the two deprivation conditions. The obtained results allowed us to conclude that the expanded time period of H2 production upon Mg-deprivation is due to the less harmful effects that Mg-depletion has on viability and metabolic performance of the cells. Unlike S-deprivation, the photosynthetic light and dark reactions in Mg-deprived cells remained active over the whole H2 production period. However, the elevated PSII activity in Mg-deprived cells was counteracted by the operation of pathways for O2 consumption that maintain anaerobic conditions in the presence of active water splitting.

Increased photosystem II stability promotes H2 production in sulfur-deprived Chlamydomonas reinhardtii.

Photobiological H2 production is an attractive option for renewable solar fuels. Sulfur-deprived cells of Chlamydomonas reinhardtii have been shown to produce hydrogen with the highest efficiency among photobiological systems. We have investigated the photosynthetic reactions during sulfur deprivation and H2 production in the wild-type and state transition mutant 6 (Stm6) mutant of Chlamydomonas reinhardtii. The incubation period (130 h) was dissected into different phases, and changes in the amount and functional status of photosystem II (PSII) were investigated in vivo by electron paramagnetic resonance spectroscopy and variable fluorescence measurements. In the wild type it was found that the amount of PSII is decreased to 25% of the original level; the electron transport from PSII was completely blocked during the anaerobic phase preceding H2 formation. This block was released during the H2 production phase, indicating that the hydrogenase withdraws electrons from the plastoquinone pool. This partly removes the block in PSII electron transport, thereby permitting electron flow from water oxidation to hydrogenase. In the Stm6 mutant, which has higher respiration and H2 evolution than the wild type, PSII was analogously but much less affected. The addition of the PSII inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea revealed that ∼80% of the H2 production was inhibited in both strains. We conclude that (i) at least in the earlier stages, most of the electrons delivered to the hydrogenase originate from water oxidation by PSII, (ii) a faster onset of anaerobiosis preserves PSII from irreversible photoinhibition, and (iii) mutants with enhanced respiratory activity should be considered for better photobiological H2 production.