Philippe Juneau

Resolution of the photosystem I and photosystem II contributions to chlorophyll fluorescence of intact leaves at room temperature

Green leaves illuminated with photosynthetically active light emit red fluorescence, whose time-dependent intensity variations reflect photosynthetic electron transport (the Kautsky effect). Usually, fluorescence variations are discussed by considering only the contribution of PSII-associated chlorophyll a, although it is known that the fluorescence of PSI-associated chlorophyll a also contributes to the total fluorescence [Aust. J. Plant Physiol. 22 (1995) 131]. Because the fluorescence emitted by each photosystem cannot be measured separately by selecting the emission wavelength in in vivo conditions, the contribution of PSI to total fluorescence at room temperature is still in ambiguity. By using a diode array detector, we measured fluorescence emission spectra corresponding to the minimal (F(O)) and maximal (F(M)) fluorescence states. We showed that the different shapes of these spectra were mainly due to a higher contribution of PSI chlorophylls in the F(O) spectrum. By exciting PSI preferentially, we recorded a reference PSI emission spectrum in the near far-red region. From the F(O) and F(M) spectra and from this PSI reference spectrum, we derived specific PSI and PSII emission spectra in both the F(O) and F(M) states. This enables to estimate true value of the relative variable fluorescence of PSII, which was underestimated in previous works. Accurate separation of PSI-PSII fluorescence emission spectra will also enable further investigations of the distribution of excitation energy between PSI and PSII under in vivo conditions.

Toxic effects and bioaccumulation of carbamazepine evaluated by biomarkers measured in organisms of different trophic levels

The antiepileptic drug carbamazapine (CBZ) readily persists in sewage-water treatment plant wastewaters and finds its way into receiving water bodies. Our study sought to examine the bioaccumulation and toxicity of CBZ using an experimental aquatic trophic chain composed of the green alga, Pseudokirchneriella subcapitata, the crustacean, Thamnocephalus platyurus, and the cnidarian, Hydra attenuata. Bioaccumulation of CBZ was estimated by liquid chromatography-tandem mass spectrometry and revealed bioaccumulation factors of 2.2 and 12.6, respectively, in algae and crustaceans. No significant bioaccumulation was observed in H. attenuata. In T. platyurus, a strong stimulation of global heme oxidase (HO) (76%), and glutathione-S-transferase activity (130%) but a drastic inhibition of cytochrome P450 3A-like activity was found which suggests alteration of enzyme activity by CBZ. However, in the hydranth H. attenuata, an increase in both global cytochrome and cytochrome P450 3A-like activity was found, while GST activity was inhibited. Lipid peroxidation was reduced in T. platyurus and H. attenuata suggesting that redox activity of the lipophilic CBZ was at play. This study highlighted the processes of carbamazepine toxicity transfer between trophic levels in aquatic organisms.

Temperature-dependent sensitivity of growth and photosynthesis of Scenedesmus obliquus, Navicula pelliculosa and two strains of Microcystis aeruginosa to the herbicide atrazine.

The temperature-dependent sensitivities of two algal species and two strains of cyanobacteria to the photosynthesis-inhibiting herbicide atrazine were evaluated in order to understand how the interaction between acclimation temperature and herbicide will affect growth and photosynthesis of aquatic microorganisms. The green alga Scenedesmus obliquus, the diatom Navicula pelliculosa and a toxic and non-toxic strain of Microcystis aeruginosa were acclimated to three different temperatures (10, 15 and 25°C) and exposed to five concentrations of the herbicide atrazine (0-0.15μM) for 72h. Growth, photosynthetic yields, energy fluxes within photosystem II and pigment content were then measured as potential responses to each treatment. With the exception of N. pelliculosa, the toxicity of atrazine was higher when microorganisms were acclimated to lower temperatures. N. pelliculosa was not only the most tolerant to atrazine, but also had a similar sensitivity to this herbicide at every temperature. The observed differences in growth sensitivity to atrazine at low temperature are associated with the ability of algae and cyanobacteria to cope with high excitation pressure, by increasing its protective carotenoid content and non-photochemical energy dissipation. Our results demonstrate that future guidelines for the protection of aquatic life should consider water temperature as an important factor influencing the toxicity of atrazine to aquatic microorganisms.

Effect of endocrine disrupters on photosystem II energy fluxes of green algae and cyanobacteria.

Among the numerous toxics found in the aquatic environment, endocrine disrupters can interfere with the normal functioning of the endocrine system of several organisms, leading to important consequences. Even if algae and cyanobacteria are non-target organisms without endocrine system, our goals were to verify if endocrine disrupters can affect photosynthetic activity and how energy flows through photosystem II (PSII) were altered. To reach these objectives, we exposed, for 15 min, two green algae (Chlamydomonas reinhardtii strain CC125, Pseudokirchneriella subcapitata strain CPCC37) and a toxic and a non-toxic strain of Microcystis aeruginosa (CPCC299 and CPCC632 respectively) to 4-octylphenol, 4-nonylphenol and β-estradiol at concentrations ranging from 0.1 to 5 μg/mL. We have shown for the first time that endocrine disrupters may have drastic effects on PSII energy fluxes. Furthermore, we showed that various species have different sensitivity to endocrine disrupters. P. subcapitata was tolerant to each endocrine disrupter tested, while flows of energy through PSII were affected similarly, but at different extent, for the other species. Cyanobacterial PSII energy fluxes were more affected than green algae, suggesting that the prokaryotic characteristics of these organisms are responsible of their high sensitivity.

Alteration of plant physiology by glyphosate and its by-product aminomethylphosphonic acid: an overview

It is generally claimed that glyphosate kills undesired plants by affecting the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme, disturbing the shikimate pathway. However, the mechanisms leading to plant death may also be related to secondary or indirect effects of glyphosate on plant physiology. Moreover, some plants can metabolize glyphosate to aminomethylphosphonic acid (AMPA) or be exposed to AMPA from different environmental matrices. AMPA is a recognized phytotoxin, and its co-occurrence with glyphosate could modify the effects of glyphosate on plant physiology. The present review provides an overall picture of alterations of plant physiology caused by environmental exposure to glyphosate and its metabolite AMPA, and summarizes their effects on several physiological processes. It particularly focuses on photosynthesis, from photochemical events to C assimilation and translocation, as well as oxidative stress. The effects of glyphosate and AMPA on several plant physiological processes have been linked, with the aim of better understanding their phytotoxicity and glyphosate herbicidal effects.

Use of chlorophyll fluorescence as a tool for determination of herbicide toxic effect: Review

Chlorophyll fluorescence-based methods have been used for many years to study the effect of environmental factors (including herbicides) on plants, and, with the development of very sensitive fluorometers, these methods have become very useful in ecotoxicological studies. This review provides a summary of the research done over the last 10 years concerning the study, by using chlorophyll fluorescence, of the herbicide toxic effects. Although numerous parameters are available, it is striking to notice that the majority of the studies done in the recent years used only a limited number of parameters, thus restraining in some way the interpretation of the obtained results. Furthermore, the use of the maximal photosystem II quantum yield to evaluate toxic effect of herbicides is probably not the best parameter due to its relatively low sensitivity. One should take the advantage of using other parameters such as the operational PSII quantum yield and the non-photochemical quenching since they integrate the entire physiological state of the plant and therefore should be more sensitive. We also recommend the systematic determination of EC50 values in future studies since such determination will permit a more accurate comparison between published reports.

Evidence for the Rapid Phytotoxicity and Environmental Stress Evaluation Using the PAM Fluorometric Method: Importance and Future Application

Modulated fluorometry has opened new possibilities for the interpretation of photosynthetic parameters concerning the physiological state of plants. By exposing plant to continuous actinic light and to pulses of saturating light, it is possible to calculate from the Kautsky fluorescence transient four important values: ΦM, the maximum quantum yield as a measure of the maximum photosystem II (PSII) photochemistry; Φ′M, the steady state quantum yield as a real PSII photochemistry at the equilibrium state of electron transport; QP, the photochemical quenching as a measure of light-energy dissipation via photosynthetic electron transport; and QN, the non-photochemical quenching as a measure of dissipation of the absorbed light energy via non-photochemical processes. The change of these PAM-fluorescence parameters is related to the physiological state of the plant and therefore they can be used as reliable indicators of different environmental stress effects. However, these parameters have never been seen as useful standard tools for toxicological monitoring of ecosystems. Highly sensitive modulated fluorometers are today available which allow to measure the fluorescence parameters either in the lab or in the field. In this report we evaluate the use of these parameters in assessing the rapid environmental impact of mercury toxicity on Selenastrum capricornutum and of temperature stress on tomato plants. In interpreting our results, we suggest the utility of some of these parameters as technologically advanced tools in future bioassays.

Differential effects of glyphosate and aminomethylphosphonic acid (AMPA) on photosynthesis and chlorophyll metabolism in willow plants

We used a willow species (Salix miyabeana cultivar SX64) to examine the differential secondary-effects of glyphosate and aminomethylphosphonic acid (AMPA), the principal glyphosate by-product, on chlorophyll metabolism and photosynthesis. Willow plants were treated with different concentrations of glyphosate (equivalent to 0, 1.4, 2.1 and 2.8kgha(-1)) and AMPA (equivalent to 0, 0.28, 1.4 and 2.8kgha(-1)) and evaluations of pigment contents, chlorophyll fluorescence, and oxidative stress markers (hydrogen peroxide content and antioxidant enzyme activities) in leaves were performed after 12h of exposure. We observed that AMPA and glyphosate trigger different mechanisms leading to decreases in chlorophyll content and photosynthesis rates in willow plants. Both chemicals induced ROS accumulation in willow leaves although only glyphosate-induced oxidative damage through lipid peroxidation. By disturbing chlorophyll biosynthesis, AMPA induced decreases in chlorophyll contents, with consequent effects on photosynthesis. With glyphosate, ROS increases were higher than the ROS-sensitive threshold, provoking chlorophyll degradation (as seen by pheophytin accumulation) and invariable decreases in photosynthesis. Peroxide accumulation in both AMPA and glyphosate-treated plants was due to the inhibition of antioxidant enzyme activities. The different effects of glyphosate on chlorophyll contents and photosynthesis as described in the literature may be due to various glyphosate:AMPA ratios in those plants.

Differential sensitivity of five cyanobacterial strains to ammonium toxicity and its inhibitory mechanism on the photosynthesis of rice-field cyanobacterium Ge–Xian–Mi (Nostoc)

Effects of two fertilizers, NH(4)Cl and KCl, on the growth of the edible cyanobacterium Ge-Xian-Mi (Nostoc) and four other cyanobacterial strains were compared at pH 8.3+/-0.2 and 25 degrees C. Their growth was decreased by at least 65% at 10 mmol L(-1) NH(4)Cl but no inhibitory effect was observed at the same level of KCl. Meanwhile, the strains exhibited a great variation of sensitivity to NH(4)(+) toxicity in the order: Ge-Xian-Mi>Anabaena azotica FACHB 118>Microcystis aeruginosa FACHB 905>M. aeruginosa FACHB 315>Synechococcus FACHB 805. The 96-h EC(50) value for relative growth rate with regard to NH(4)(+) for Ge-Xian-Mi was 1.105 mmol L(-1), which was much less than the NH(4)(+) concentration in many agricultural soils (2-20 mmol L(-1)). This indicated that the use of ammonium as nitrogen fertilizer was responsible for the reduced resource of Ge-Xian-Mi in the paddy field. After 96 h exposure to 1 mmol L(-1) NH(4)Cl, the photosynthetic rate, F(v)/F(m) value, saturating irradiance for photosynthesis and PSII activity of Ge-Xian-Mi colonies were remarkably decreased. The chlorophyll synthesis of Ge-Xian-Mi was more sensitive to NH(4)(+) toxicity than phycobiliproteins. Thus, the functional absorption cross section of Ge-Xian-Mi PSII was increased markedly at NH(4)Cl levels >or=1 mmol L(-1) and the electron transport on the acceptor side of PSII was significantly accelerated by NH(4)Cl addition >or=3 mmol L(-1). Dark respiration of Ge-Xian-Mi was significantly increased by 246% and 384% at 5 and 10 mmol L(-1) NH(4)Cl, respectively. The rapid fluorescence rise kinetics indicated that the oxygen-evolving complex of PSII was the inhibitory site of NH(4)(+).

Dichromate effect on energy dissipation of photosystem II and photosystem I in Chlamydomonas reinhardtii

In this study, we investigated the energy dissipation processes via photosystem II and photosystem I activity in green alga Chlamydomonas reinhardtii exposed to dichromate inhibitory effect. Quantum yield of photosystem II and also photosystem I were highly decreased by dichromate effect. Such inhibition by dichromate induced strong quenching effect on rapid OJIP fluorescence transients, indicating deterioration of photosystem II electron transport via plastoquinone pool toward photosystem I. The decrease of energy dissipation dependent on electron transport of photosystem II and photosystem I by dichromate effect was associated with strong increase of non-photochemical energy dissipation processes. By showing strong effect of dichromate on acceptor side of photosystem I, we indicated that dichromate inhibitory effect was not associated only with PSII electron transport. Here, we found that energy dissipation via photosystem I was limited by its electron acceptor side. By the analysis of P700 oxido-reduction state with methylviolagen as an exogenous PSI electron transport mediator, we showed that PSI electron transport discrepancy induced by dichromate effect was also caused by inhibitory effect located beyond photosystem I. Therefore, these results demonstrated that dichromate has different sites of inhibition which are associated with photosystem II, photosystem I and electron transport sink beyond photosystems.