Dichlorophenyl ) -1,1-dimethylurea Research Articles

Biochemical and molecular characterization of the SBiP1 chaperone from Symbiodinium microadriaticum CassKB8 and light parameters that modulate its phosphorylation.

The coding and promoter region sequences from the BiP-like protein SBiP1 from Symbiodinium microadriaticum CassKB8 were obtained by PCR, sequenced and compared with annotated sequences. The nucleotides corresponding to the full sequence were correctly annotated and the main SBiP1 features determined at the nucleotide and amino acid level. The translated protein was organized into the typical domains of the BiP/HSP70 family including a signal peptide, a substrate- and a nucleotide-binding domain, and an ER localization sequence. Conserved motifs included a highly conserved Thr513 phosphorylation site and two ADP-ribosylation sites from eukaryotic BiP's. Molecular modeling showed the corresponding domain regions and main exposed post-translational target sites in its three-dimensional structure, which also closely matched Homo sapiens BiP further indicating that it indeed corresponds to a BiP/HSP70 family protein. The gene promoter region showed at least eight light regulation-related sequences consistent with the molecule being highly phosphorylated in Thr under dark conditions and dephosphorylated upon light stimuli. We tested light parameter variations that could modulate the light mediated phosphorylation effect and found that SBiP1 Thr dephosphorylation was only significantly detected after 15-30 min light stimulation. Such light-induced dephosphorylation was observed even when dichlorophenyl dimethyl urea, a photosynthesis inhibitor, was also present in the cells during the light stimulation. Dephosphorylation occurred indistinctly under red, yellow, blue or the full visible light spectra. In additon, it was observed at a light intensity of as low as 1 μmole photon m-2 s-1. Our results indicate that: a) SBiP1 is a chaperone belonging to the BiP/HSP70 family proteins; b) its light-modulated phosphorylation/dephosphorylation most likely functions as an activity switch for the chaperone; c) this light-induced modulation occurs relatively slow but is highly sensitive to the full spectrum of visible light; and d) the light induced Thr dephosphorylation is independent of photosynthetic activity in these cells.

Quantitative Representation of Water Quality Biotoxicity by Algal Photosynthetic Inhibition.

The method based on the photosynthetic inhibition effect of algae offers the advantages of swift response and straightforward measurement. Nonetheless, this effect is influenced by both the environment and the state of the algae themselves. Additionally, a single parameter is vulnerable to uncertainties, rendering the measurement accuracy and stability inadequate. This paper employed currently utilized photosynthetic fluorescence parameters, including Fv/Fm(maximum photochemical quantum yield), Performance Indicator (PIabs), Comprehensive Parameter Index (CPI) and Performance Index of Comprehensive Toxicity Effect (PIcte), as quantitative toxicity characteristic parameters. The paper compared the univariate curve fitting results with the multivariate data-driven model results and investigated the effectiveness of Back Propagation(BP) Neural Network and Support Vector Machine for Regression (SVR) models to enhance the accuracy and stability of toxicity detection. Using Dichlorophenyl Dimethylurea (DCMU) samples as an example, the mean Relative Root Mean Square Error (RRMSE) corresponding to the optimal parameter PIcte for the dose-effect curve fitting was 1.246 in the concentration range of 1.25-200 µg/L. On the other hand, the mean RRMSEs corresponding to the results of the BP neural network and SVR models were 0.506 and 0.474, respectively. Notably, BP neural network exhibited excellent prediction accuracy in the medium-high concentration range of 7.5-200 µg/L, with a mean RRSME of only 0.056. Regarding the stability of the results, the mean Relative Standard Deviation (RSD) of the univariate dose-effect curve results was 15.1% within the concentration range of 50-200 µg/L. In contrast, the mean RSDs for both BP neural network and SVR results were less than 5%. In the concentration range of 1.25-200 µg/L, the mean RSDs were 6.1% and 16.5%, with the BP neural network performing well. The experimental results of Atrazine were analyzed to further validate the effectiveness of the BP neural network in improving the accuracy and stability of results. These findings provided valuable insights for the development of biotoxicity detection by using the algae photosynthetic inhibition method.

Time course of the photochemical reflectance index during photosynthetic induction: its relationship with the photochemical yield of photosystem II.

Time courses of photochemical reflectance index (PRI) of an attached cucumber leaf during a dark-light transition were compared with those of photochemical yields of photosystem II (YII ) to discuss the feasibility of PRI imaging for estimating the efficiency of photosynthetic light use. YII and PRI were simultaneously evaluated with a pulse-amplitude modulation chlorophyll fluorometer and a low-cost imaging system consisting of digital cameras and band-pass filters, respectively. YII decreased immediately after the transition and then increased under various photon flux densities. Although PRI exhibited delayed time courses with respect to YII under low light conditions, PRI decreased monotonically under high light conditions. There was no correlation between YII and the changes in PRI (ΔPRI) immediately after the transition but YII was correlated with ΔPRI under the steady-state photosynthesis. These results indicate that the use of PRI to estimate YII under fluctuating light based on the regression obtained at steady state can overestimate YII . The imaging system was also applied to evaluate the spatial PRI distribution within a leaf. While PRI of leaf areas that remained untreated, or had been treated with H2 O again, first dropped and then rose under low light and monotonically decreased under high light conditions, leaf areas treated with inhibitor (dichlorophenyl dimethylurea) did not exhibit any changes. It is likely that the inhibitor suppressed lumen acidification, which triggers a decrease in PRI. It was suggested that YII of leaves with malfunctions in the photosynthetic electron transport can be overestimated by the PRI-based estimation.

Spectrally decomposed dark-to-light transitions in Synechocystis sp. PCC 6803

Photosynthetic activity and respiration share the thylakoid membrane in cyanobacteria. We present a series of spectrally resolved fluorescence experiments where whole cells of the cyanobacterium Synechocystis sp. PCC 6803 and mutants thereof underwent a dark-to-light transition after different dark-adaptation (DA) periods. Two mutants were used: (i) a PSI-lacking mutant (ΔPSI) and (ii) M55, a mutant without NAD(P)H dehydrogenase type-1 (NDH-1). For comparison, measurements of the wild-type were also carried out. We recorded spectrally resolved fluorescence traces over several minutes with 100 ms time resolution. The excitation light was at 590 nm so as to specifically excite the phycobilisomes. In ΔPSI, DA time has no influence, and in dichlorophenyl-dimethylurea (DCMU)-treated samples we identify three main fluorescent components: PB–PSII complexes with closed (saturated) RCs, a quenched or open PB–PSII complex, and a PB–PSII ‘not fully closed.’ For the PSI-containing organisms without DCMU, we conclude that mainly three species contribute to the signal: a PB–PSII–PSI megacomplex with closed PSII RCs and (i) slow PB → PSI energy transfer, or (ii) fast PB → PSI energy transfer and (iii) complexes with open (photochemically quenched) PSII RCs. Furthermore, their time profiles reveal an adaptive response that we identify as a state transition. Our results suggest that deceleration of the PB → PSI energy transfer rate is the molecular mechanism underlying a state 2 to state 1 transition.

Diverse responses of Symbiodinium types to menthol and DCMU treatment.

To understand the mechanism of photosynthetic inhibition and generation of reactive oxygen species (ROS) in Symbiodinium types under stress, chemicals such as dichlorophenyl dimethylurea (DCMU) are widely used. Moreover, DCMU and recently menthol were used to generate aposymbiotic cnidarian hosts. While the effects of DCMU on Symbiodinium cells have been extensively studied, no studies have shown the mechanism behind menthol-induced coral bleaching. Moreover, no study has compared the effects of DCMU and menthol treatments on photosystem II (PSII) activity and generation of ROS in different Symbiodinium types. In this study, we utilized five freshly isolated Symbiodinium types (S. minutum (B1), S. goreaui (C1), C3, C15, and S. trenchii (D1a)) to compare the effects of DCMU and menthol treatments. Symbiodinium cells were exposed to DCMU and menthol at different concentrations for 4 h. Results showed that values of the 50% inhibitory concentration (IC50) for PSII inhibition were 0.72∼1.96 mM for menthol-treated cells compared to 29∼74 pM for DCMU-treated cells. Diverse responses of Symbiodinium types were displayed in terms of PSII tolerance to menthol (S. minutum > S. trenchii = C15 > C3 = S. goreaui), and also in the response curves. In contrast, responses were not so diverse when the different types were treated with DCMU. Three of five menthol-treated Symbiodinium types showed instant and significant ROS generation when PSII activity was inhibited, compared to no ROS being generated in DCMU-treated Symbiodinium types. Both results indicated that menthol inhibited Symbiodinium PSII activity through Symbiodinium type-dependent mechanisms, which were also distinct from those with DCMU treatment. This study further confirmed that photosynthetic functions Symbiodinium have diverse responses to stress even within the same clade.

Enhanced production of fatty acids in three strains of microalgae using a combination of nitrogen starvation and chemical inhibitors of carbohydrate synthesis

Microalgae are attracting much attention as superior biodiesel producers. In particular, under stressful conditions, they accumulate organic compounds consisting entirely of carbon and hydrogen. The aim of this work was to increase intracellular fatty acid content in Dunaliella tertiolecta (Chlorophyceae), Nannochloropsis oculata (Eustigmatophyceae), and Porphyridium cruentum (Rhodophyceae) using a combination of nitrogen starvation and chemical inhibitors of carbohydrate biosynthesis. These microalgae were subjected to nitrogen starvation and their physiological changes were then observed over time. In D. tertiolecta, no significant change in total fatty acid content was detected on day 3.5 relative to the initial total fatty acid content (day 0), while total carbohydrate content dramatically increased as the nitrogen starvation period was extended. In N. oculata, total fatty acid content rapidly increased, reaching up to nearly 40% of the DCW at day 3.5. However, total carbohydrate content exhibited a gradual reduction throughout the experiment. In P. cruentum, total carbohydrate content increased up to 43% of DCW on day 3.5 and total fatty acid content increased slightly under nitrogen depletion. These data suggest that different eukaryotic microalgae use different storage products under stressful conditions. Among the three strains, D. tertiolecta showed decreased total carbohydrate content and enhanced total fatty acid content following inhibition of carbohydrate synthesis by dichlorophenyl dimethylurea and cyclohexane diamine tetra acetic acid. The results demonstrate the possibility of furthering our understanding of the fatty acid and carbohydrate biosynthesis metabolic network that responds to environmental changes in microalgae.

The reduced state of the plastoquinone pool is required for chloroplast‐mediated stomatal closure in response to calcium stimulation

Besides their participation in photosynthesis, leaf chloroplasts function in plant responses to stimuli, yet how they direct stimulus-induced stomatal movement remains elusive. Here, we showed that over-reduction of the plastoquinone (PQ) pool by dibromothymoquinone (DBMIB) was closely associated with stomatal closure in plants which required chloroplastic H2O2 generation in the mesophyll. External application of H2 O2 reduced the PQ pool, whereas the cell-permeable reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) reversed the DBMIB-induced over-reduction of the PQ pool and stomatal closure. Mesophyll chloroplasts are key players of extracellular Ca(2+) (Ca(2+)o)-induced stomatal closure, but when treated with either 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) or NAC they failed to facilitate Ca(2+)o-induced stomatal closure due to the inhibition of chloroplastic H2 O2 synthesis in mesophyll. Similarly, the Arabidopsis electron transfer chain-related mutants npq4-1, stn7 and cas-1 exhibited diverse responses to Ca(2+)o or DBMIB. Transcriptome analysis also demonstrated that the PQ pool signaling pathway shared common responsive genes with the H2 O2 signaling pathway. These results implicated a mechanism for chloroplast-mediated stomatal closure involving the generation of mesophyll chloroplastic H2O2 based on the reduced state of the PQ pool, which is calcium-sensing receptor (CAS) and LHCII phosphorylation dependent.

The sporulation of the green alga Ulva prolifera is controlled by changes in photosynthetic electron transport chain.

Sporulation and spore release are essential phases of the life cycle in algae and land plants. Ulva prolifera, which is an ideal organism for studying sporulation and spore release, was used as the experimental material in the present study. The determination of photosynthetic parameters, combined with microscopic observation, treatment with photosynthetic inhibitors, limitation of carbon acquisition, and protein mass spectrometry, was employed in this experiment. Cycle electron transport (CEF) was found enhanced at the onset of sporangia formation. The inhibition effect of dibromothymoquinone (DBMIB) towards sporulation was always strong during the sporulation process whereas the inhibition effect of 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU) was continuously declined accompanied with the progress of sporulation. The changes of photosynthesis resulted from the limitation of CO2 acquisition could stimulate sporulation onset. Quantitative protein analysis showed that enzymes involved in carbon fixation, including RUBISCO and pyruvate orthophosphate dikinase, declined during sporogenesis, while proteins involved in sporulation, including tubulin and centrin, increased. These results suggest that enhanced cyclic electron flow (CEF) and oxidation of the plastoquinone pool are essential for sporangia formation onset, and changes in photosynthetic electron transport chain have significant impacts on sporulation of the green algae.

NADPH from the oxidative pentose phosphate pathway drives the operation of cyclic electron flow around photosystem I in high-intertidal macroalgae under severe salt stress.

Pyropia yezoensis (Bangiales, Rhodophyta) is a representative species of high-intertidal macroalgae, whose blades can tolerate extreme stresses, such as salt stress and desiccation. In this study, the photosystem (PS) responses of P. yezoensis blades under salt stress were studied. Our results showed that when the effective photochemical quantum yield of PS (Y) II decreased to almost zero under high salt stress, YI still had a relatively high activity rate. PSII was therefore more sensitive to salt stress than PSI. Furthermore, in the presence of 3-(3', 4'-dichlorophenyl)-1,1-dimethylurea (DCMU), YI rose as salinity increased. The YI values for DCMU-treated thalli decreased in the presence of glucose-6-phosphate dehydrogenase (EC 1.1.1.49, G6PDH) inhibitor (glucosamine, Glucm). The YI values were ∼0.09 in the presence of methyl viologen (MV) and almost zero in the presence of dibromothymoquinone (DBMIB). These results demonstrated that under severe salt stress (120‰ salinity) PSI activity was driven from a source other than PSII, and that stromal reductants probably supported the operation of PSI. Under salt stress, the starch content decreased and soluble sugar levels increased. The G6PDH and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) activities increased, but cytosolic glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12) activity decreased. Furthermore, the NADPH content increased, but NADH decreased, which suggested that soluble sugar entered the oxidative pentose phosphate pathway (OPPP). All these results suggested that NADPH from OPPP increases the cyclic electron flow around PSI in high-intertidal macroalgae under severe salt stress.

Regulation of iron-siderophore uptake in a diazotrophic cyanobacterium Anabaena anabaena 7120

The regulation of 55Fe-siderophore uptake of was studied in a diazotrophic cyanobacterium Anabaena 7120. The uptake up to 20 min was found to be biphasic, a rapid first and steady second phase. A single kinetic system mediated the uptake of iron-siderophore with an apparent Km of 200 µM and a Vmax of 20 nmol iron taken up mg-1 protein min-1. 3-(3,4dichlorophenyl)-1, 1-dimethyl urea (DCMU), 2-n-heptyl-4-hydroxy quinoline (HOQNO), dicyclohexyl carbodiimide (DCCD), carbonyl cyanide p-fluoromethoxy phenyl hydrazone (FCCP) and carbonyl cyanide-m-chlorophenyl hydrazone (CCCP) inhibited the iron-siderophore uptake significantly, whereas KCN inhibited marginally. No iron-siderophore uptake was observed under dark-aerobic and dark-anaerobic conditions. The siderophore schizokinen supported the maximum iron-siderophore uptake. However, the siderophore aerobactin and desferal could support only 53 and 3.4% iron-siderophore uptake, respectively as compared to its schizokinen counterpart.

The physiological links of the increased photosystem II activity in moderately desiccated Porphyra haitanensis (Bangiales, Rhodophyta) to the cyclic electron flow during desiccation and re-hydration

Photosynthetic electron flow changed considerably during desiccation and re-hydration of the intertidal macroalgae Porphyra haitanensis. Activities of both photosystem (PSI) and photosystem (PSII) increased significantly at moderate desiccation levels. Whereas PSII activity was abolished at an absolute water content (AWC) <24%, PSI remained active with progressive decreases in AWC to values as low as 16%. This result suggested that cyclic electron flow around PSI was still active after inactivation of linear electron flow following severe desiccation. Moreover, the PSI activity was restored more rapidly than that of PSII upon re-hydration. Pretreatment of the blades with 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) suppressed PSII activity following desiccation to an AWC of ~16% AWC. Cyclic electron flow around PSI decreased markedly in blades pretreated with DCMU than in blades without pretreatment of DCMU during re-hydration in seawater containing DCMU. All results suggested that the activity of PSII under desiccation conditions plays an important role in the operation of cyclic electron flow during desiccation and its recovery during re-hydration. Therefore, we proposed the PSII activity during desiccation could eventually lead to the accumulation of NADPH, which could serve as electron donor for P700(+) and promote its recovery during re-hydration, thereby favoring the operation of cyclic electron flow.

Evidence for a fluorescence yield change driven by a light-induced conformational change within photosystem II during the fast chlorophyll a fluorescence rise

Experiments were carried out to identify a process co-determining with Q A the fluorescence rise between F 0 and F M. With 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU), the fluorescence rise is sigmoidal, in its absence it is not. Lowering the temperature to − 10 °C the sigmoidicity is lost. It is shown that the sigmoidicity is due to the kinetic overlap between the reduction kinetics of Q A and a second process; an overlap that disappears at low temperature because the temperature dependences of the two processes differ. This second process can still relax at − 60 °C where recombination between Q A − and the donor side of photosystem (PS) II is blocked. This suggests that it is not a redox reaction but a conformational change can explain the data. Without DCMU, a reduced photosynthetic electron transport chain (ETC) is a pre-condition for reaching the F M. About 40% of the variable fluorescence relaxes in 100 ms. Re-induction while the ETC is still reduced takes a few ms and this is a photochemical process. The fact that the process can relax and be re-induced in the absence of changes in the redox state of the plastoquinone (PQ) pool implies that it is unrelated to the Q B-occupancy state and PQ-pool quenching. In both +/−DCMU the process studied represents ~ 30% of the fluorescence rise. The presented observations are best described within a conformational protein relaxation concept. In untreated leaves we assume that conformational changes are only induced when Q A is reduced and relax rapidly on re-oxidation. This would explain the relationship between the fluorescence rise and the ETC-reduction.

Photoinactivation of photosystem II: is there more than one way to skin a cat?

We briefly review the main mechanisms proposed for photodamage to photosystem II (PSII), at the donor and acceptor sides, and then discuss the mechanism whereby filamentous cyanobacteria inhabiting biological sand crusts such as Microcoleus sp. are able to avoid serious damage to their photosynthetic machinery. We show that the decline in fluorescence following exposure to excess light does not reflect a reduction in PSII activity but rather the activation of a non-radiative charge recombination in PSII. Furthermore, we show that the difference in the thermoluminescent peak temperature intensities in these organisms, in the presence and absence of inhibitors such as dichlorophenyl-dimethylurea (DCMU), is smaller than observed in model organisms suggesting that the redox gap between Q(A)⁻ and P₆₈₀+ is smaller. On the basis of these data, we propose that this could enable an alternative, pheophytin-independent recombination, thereby minimizing the damaging ¹O₂ production associated with radiative recombination.

ON THE ROLE OF THE CYTOSKELETON IN COCCOLITH MORPHOGENESIS: THE EFFECT OF CYTOSKELETON INHIBITORS1

The coccolithophore Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler was cultured in natural seawater with the addition of either the microtubule‐inhibitor colchicine, the actin‐inhibitor cytochalasin B, or the photosynthesis inhibitor 3‐(3,4 dichlorophenyl)‐1,1‐dimethyl‐urea (DCMU). Additionally, E. huxleyi was cultured at different light intensities and temperatures. Growth rate was monitored, and coccolith morphology analyzed. While every treatment affected growth rate, the percentage of malformed coccoliths increased with colchicine, cytochalasin B, and at higher than optimal temperature. These results represent the first experimental evidence for the role of microtubules and actin microfilaments in coccolith morphogenesis.

Light-dependent adsorption of photosynthetic cyanophages to Synechococcus sp. WH7803

Cyanophages infecting marine Synechococcus strains are abundant in the world's oceans and are of considerable ecological significance by virtue of their hosts' role as prominent primary producers in the marine environment. In nature, cyanobacteria experience diel light-dark (LD) cycles, which may exert significant effects on the phage life cycle. An investigation into the role of light revealed that cyanophage S-PM2 adsorption to Synechococcus sp. WH7803 was a light-dependent process. Phage adsorption assays were carried out under illumination at different wavelengths and also in the presence of photosynthesis inhibitors. Furthermore, phage adsorption was also assayed to LD-entrained cells at different points in the circadian cycle. Cyanophage S-PM2 exhibited a considerably decreased adsorption rate under red light as compared with blue, green, yellow light or daylight. However, photosynthesis per se was not required for adsorption as inhibitors such as dichlorophenyldimethyl urea did not affect the process. Neither was S-PM2 adsorption influenced by the circadian rhythm of the host cells. The presence or absence of the photosynthetic reaction centre gene psbA in cyanophage genomes was not correlated with the light-dependent phage adsorption.

Role of Light in the Appearance of Glutamine Synthetase in Leaves of Pennisetum glaucum

Leaves of Pennisetum [Pennisetum glaucum (L) HHB 67] seedlings contained two isozymes of glutamine synthetase (GS, EC 6.3.1.2): cytosolic GS1 and chloroplastic GS2. Leaves of seedlings grown in light for seven days contained about twofold higher GS activity than etiolated leaves. In both light and dark grown seedlings, total GS, GS1 and GS2 activity declined with plant age with more pronounced effect in leaves of etiolated seedlings, and GS2 declined at a much faster rate than GS1. Exposure of etiolated seedlings to light markedly enhanced GS1 and GS2 activity. This increase in activity was not affected by cycloheximide, precluding light dependent de novo synthesis of the enzyme. Treatment of etiolated seedlings with photosynthetic inhibitor, dichlorophenyl dimethyl urea (DCMU) inhibited light dependent appearance of GS. Exogenous supply of sucrose to dark grown seedlings greatly increased the GS activity in dark. These results suggest that light-mediated stimulation in activity of GS in Pennisetum leaves is dependent on photosynthetic reaction.

Sulfide, microcystin, and the etiology of black band disease

Black band disease (BBD) consists of a cyanobacterial-dominated, sulfide-rich microbial mat that migrates across coral colonies, degrading coral tissue. The mat contains diverse bacteria that include photoautotrophs (cyanobacteria), sulfate-reducers, sulfide-oxidizers, and organoheterotrophs. BBD sulfate-reducers contribute to BBD pathobiology by production of sulfide, which causes coral tissue lysis and death, and the cyanotoxin microcystin is produced by BBD cyanobacteria. Here we used a model system of coral fragments to investigate the roles of sulfide and microcystin in BBD by exposure to the metabolic inhibitors sodium molybdate and 3-(3', 4'-dichlorophenyl)-1, 1-dimethylurea (DCMU), which inhibit sulfate reduction and oxygenic photosynthesis, respectively. Exposure of BBD inocula to sodium molybdate prior to inoculation prevented infection of healthy fragments but did not prevent continued band migration and coral tissue lysis by active BBD infections. Exposure to DCMU did not inhibit either the initiation of BBD or continued migration of active BBD. Exposure of healthy coral fragments to sulfide, purified microcystin, and a combination of both revealed that both microcystin and sulfide are toxic to coral and act synergistically. Measurement of growth of bacteria isolated from BBD and the healthy coral surface mucopolysaccharide layer (SML) during exposure to microcystin revealed that growth of relatively more BBD than SML isolates was stimulated, although effects were not uniform and the majority exhibited no effect. Our results indicate that sulfide is required for initiation of BBD, both microcystin and sulfide are involved in BBD pathobiology, and microcystin may structure the BBD bacterial community.

Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis

Oxygen-evolving Photosystem II particles (crude PSII) retaining a high oxygen-evolving activity have been prepared from a marine centric diatom, Chaetoceros gracilis (Nagao et al., 2007). The crude PSII, however, contained a large amount of fucoxanthin chlorophyll a/c-binding proteins (FCP). In this study, a purified PSII complex which was deprived of major components of FCP was isolated by one step of anion exchange chromatography from the crude PSII treated with Triton X-100. The purified PSII was still associated with the five extrinsic proteins of PsbO, PsbQ', PsbV, Psb31 and PsbU, and showed a high oxygen-evolving activity of 2135 μmol O2 (mg Chl a)−1 h−1 in the presence of phenyl-p-benzoquinone which was virtually independent of the addition of CaCl2. This activity is more than 2.5-fold higher than the activity of the crude PSII. The activity was completely inhibited by 3-(3,4)-dichlorophenyl-(1,1)-dimethylurea (DCMU). The purified PSII contained 42 molecules of Chl a, 2 molecules of diadinoxanthin and 2 molecules of Chl c on the basis of two molecules of pheophytin a, and showed typical absorption and fluorescence spectra similar to those of purified PSIIs from the other organisms. In this study, we also found that the crude PSII was significantly labile, as a significant inactivation of oxygen evolution, chlorophyll bleaching and degradation of PSII subunits were observed during incubation at 25 °C in the dark. In contrast, these inactivation, bleaching and degradation were scarcely detected in the purified PSII. Thus, we succeeded for the first time in preparation of a stable PSII from diatom cells.

3D monitoring spatio-temporal effects of herbicide on a whole plant using combined range and chlorophyll a fluorescence imaging.

Spatio-temporal effects of herbicide including 3-(3,4 dichlorophenyl)-1,1-dimethylurea (DCMU) on a whole melon (Cucumis melo L.) plant were three-dimensionally monitored using combined range and chlorophyll a fluorescence imaging. The herbicide was treated to soil in a pot and the changes in chlorophyll a fluorescence images of the plant were captured over time. The time series of chlorophyll fluorescence images were combined with 3D polygon model of the whole plant taken by a high-resolution portable scanning lidar. From the produced 3D chlorophyll fluorescence model, it was observed that the increase of chlorophyll fluorescence appeared along veins of leaves and gradually expanded to mesophylls. In addition, it was found by detailed analysis of the images that the invisible herbicide injury on the mature leaves occurred earlier and more severely than on the young and old leaves. The distance from veins, whole leaf area and leaf inclination influenced the extent of the injury within the leaves. These results indicated difference in uptake of herbicide in the plant from soil depends on structural parameters of leaves and the microenvironments as well as leaf age. The findings showed that 3D monitoring using combined range and chlorophyll a fluorescence imaging can be utilised for understanding spatio-temporal changes of herbicide effects on a whole plant.

Adaptation of cyanobacteria to the sulfide-rich microenvironment of black band disease of coral

Black band disease (BBD) is a cyanobacteria-dominated microbial mat that migrates across living coral colonies lysing coral tissue and leaving behind exposed coral skeleton. The mat is sulfide-rich due to the presence of sulfate-reducing bacteria, integral members of the BBD microbial community, and the sulfide they produce is lethal to corals. The effect of sulfide, normally toxic to cyanobacteria, on the photosynthetic capabilities of five BBD cyanobacterial isolates of the genera Geitlerinema (3), Leptolyngbya (1), and Oscillatoria (1) and six non-BBD cyanobacteria of the genera Leptolyngbya (3), Pseudanabaena (2), and Phormidium (1) was examined. Photosynthetic experiments were performed by measuring the photoincorporation of [(14)C] NaHCO(3) under the following conditions: (1) aerobic (no sulfide), (2) anaerobic with 0.5 mM sulfide, and (3) anaerobic with 0.5 mM sulfide and 10 microM 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU). All five BBD cyanobacterial isolates tolerated sulfide by conducting sulfide-resistant oxygenic photosynthesis. Five of the non-BBD cyanobacterial isolates did not tolerate sulfide, although one Pseudanabaena isolate continued to photosynthesize in the presence of sulfide at a considerably reduced rate. None of the isolates conducted anoxygenic photosynthesis with sulfide as an electron donor. This is the first report on the physiology of a culture of Oscillatoria sp. found globally in BBD.