Minor but key chlorophylls in Photosystem II

A ‘metal-free’ chlorophyll (Chl) a, pheophytin (Phe) a, functions as the primary electron acceptor in PS II. On the basis of Phe a/PS II = 2, Phe a content is postulated as an index for estimation of the stoichiometry of pigments and photosystems. We found Phe a in a Chl d-dominant cyanobacterium Acaryochloris marina, whereas Phe d was absent. The minimum Chl a:Phe a ratio was 2:2, indicating that the primary electron donor is Chl a, accessory is Chl d, and the primary electron acceptor is Phe a in PS II of A. marina. Chl d was artificially formed by the treatment of Chl a with papain in aqueous organic solvents. Further, we will raise a key question on the mechanisms of water oxidation in PS II.     

Cyanobacterial toxicity and migration in a mesotrophic lake in western Washington, USA

In fall 1997, the toxic cyanobacterium Microcystis aeruginosa was documented in Lake Sammamish (western Washington, U.S.A.) for the first time. Cyanobacterial activity and environmental conditions that may promote toxic cyanobacteria were investigated during summer and fall 1999. Development of toxic Microcystis was hypothesized to be due to runoff of nutrients from the watershed (external loading hypothesis) or from vertical migration of dormant cyanobacteria from the nutrient-rich sediments into the water column (cyanobacterial migration hypothesis). Microcystins were detected using an enzyme-linked immunosorbent assay during late August and early September 1999 despite low cyanobacterial abundance. Microcystin concentrations ranged between 0.19–3.8 g l^–1 throughout the lake and at all depths with the exception of the boat launch where concentrations reached 43 g l^–1. Comparison of the conditions associated with the toxic episodes in 1997 and 1999 indicate that Microcystis is associated with a stable water column, increased surface total phosphorus concentrations (> 10 g l^–1), surface temperatures greater than 22°C, high total nitrogen to phosphorus ratios (> 30), and increased water column transparency (up to 5.5 m). Migration of the cyanobacteria, Microcystis and Anabaena, occurred in both the deep and shallow portions of the lake. Microcystis dominated (89–99%) the migrating cyanobacteria with greater migration from the shallow station. External loading of nutrients due to the large rainfall preceding the 1997 toxic episode may have provided the nutrients needed to fuel that bloom. However, toxic Microcystis occurred in 1999 despite the lack of rain and subsequent external runoff. The migration of Microcystis from the nutrient-rich sediments may have been the inoculum for the toxic population detected in 1999.     

Daily changes of uptake of inorganic carbon and nitrogen,and their relation to phytoplankton blooms in late spring-early summer in Lake Nakanuma,Japan

Daily changes of inorganic carbon and nitrogen uptake were measured in May in 1986 in Lake Nakanuma, Japan. Uptake of inorganic carbon and ammonium in the light-bottle experiments in the 1 m layers, showed daily changes similar to chlorophyll a changes, though the uptake activities peaked before chlorophyll a peaks (phytoplankton blooms) appeared. Potential growth rates of phytoplankton and observed growth rates were calculated from the uptake rates and chlorophyll a changes. The potential growth rates did not always correspond to the observed growth rates. The potential growth rates did not correlate with the loss rates. The correlation between the observed growth rates and the loss rates was better. These results suggest that though the increase of uptake activities may be necessary for occurrence of phytoplankton blooms, loss processes may affect the occurrence of blooms.     

Textile dye decolorization using cyanobacteria

Cyanobacterial cultures isolated from sites polluted by industrial textile effluents were screened for their ability to decolorize cyclic azo dyes. Gloeocapsa pleurocapsoides and Phormidium ceylanicum decolorized Acid Red 97 and FF Sky Blue dyes by more than 80% after 26 days. Chroococcus minutus was the only culture which decolorized Amido Black 10B (55%). Chlorophyll a synthesis in all cultures was strongly inhibited by the dyes. Visible spectroscopy and TLC confirmed that color removal was due to degradation of the dyes.Revisions requested 10 November 2004/30 November 2004; Revisions received 16 November 2004/ 7 January 2005     

Which side of the π-macrocycle plane of (bacterio)chlorophylls is favored for binding of the fifth ligand?

Reported crystallographic data and calculated molecular models indicated that chlorophyll (Chl) a and bacteriochlorophyll (BChl) a tend to bind the fifth ligand on the side of the macrocycle where the C13²-(R)-methoxycarbonyl moiety protrudes (denoting the ‘back’ side). The crystal structures of 34 photosynthetic proteins possessing (B)Chl cofactors revealed that most of Chl a and BChl a (and b) are coordinated by any peptidyl residue (e.g., histydyl-imidazolyl group), peptidyl backbone or water from the ‘back’ side. Almost all the cofactors that bind a water molecule as the fifth ligand in these proteins have a ‘back’ configuration. Theoretical model calculations for methyl chlorophyllide a (MeChlid a) and methyl bacteriochlorophyllide a (MeBChlid a) bound to an imidazole molecule indicated that the ‘back’ side is energetically favored for the ligand binding. These results are consistent with the fact that ethyl chlorophyllide a (EtChlid a) dihydrate crystal consists of the ‘back’ complex. The modeling also showed that both removal and stereochemical inverse of the C13²-methoxycarbonyl group affect the relative stability between the ‘back’ and ‘face’ complexes. The effect of the C13²-moiety on the choice of the macrocycle side for the ligand binding is discussed in relation to the function of P700. This revised version was published online in June 2006 with corrections to the Cover Date.     

Phytoplankton growth rates in the freshwater tidal reaches of the Schelde estuary (Belgium) estimated using a simple light-limited primary production model

During the course of 1996, phytoplankton was monitored in the turbid, freshwater tidal reaches of the Schelde estuary. Using a simple light-limited primary production model, phytoplankton growth rates were estimated to evaluate whether phytoplankton could attain net positive growth rates and whether growth rates were high enough for a bloom to develop. Two phytoplankton blooms were observed in the freshwater tidal reaches. The first bloom occurred in March and was mainly situated in the most upstream reaches of the freshwater tidal zone, suggesting that it was imported from the tributary river Schelde. The second bloom occurred in July and August. This summer bloom was situated more downstream in the freshwater tidal reaches and appeared to have developed within the estuary. A comparison between phytoplankton growth rates estimated using a simple primary production model and flushing rate of the water indicated that no net increase in phytoplankton biomass was possible in March while phytoplankton could theoretically increase its biomass by 20% per day during summer. Chlorophyllaconcentrations at all times decreased strongly at salinities between 5–10 psu. This decline was ascribed to a combination of salinity stress and light limitation. Phytoplankton biomass and estimated annual net production were much higher in the freshwater tidal zone compared to the brackish reaches of the estuary (salinity > 10 psu) despite mixing depth to euphotic depth ratios being similar. Possible reasons for this high production include high nutrient concentrations, low zooplankton grazing pressure and import of phytoplankton blooms from the tributary rivers.     

Potassium-induced inhibition of nitrogen and phosphorus metabolism as a strategy of controlling <Emphasis Type="Italic">Microcystis</Emphasis> blooms

To explore potassium toxicity in Microcystis sp., growth, chlorophyll a, carotenoid and phycocyanin content, uptake of nitrate, phosphate and ammonium and activities of the assimilatory enzymes nitrate reductase, alkaline phosphatase and glutamine synthetase (GS) were studied. Nitrate, phosphate, ammonium uptakes and chlorophyll a and phycocyanin contents decreased with increase in the concentration of potassium, but carotenoid content registered an increase at increasing potassium concentration. Alkaline phosphatase and GS activities followed the trend of inhibition of their respective nutrients, whereas nitrate and nitrate reductase showed negative correlation (p < 0.01). Potassium was found to inhibit the activities of all the assimilatory enzymes in a non-competitive manner. Inhibitions of these parameters support the view that potassium has the potential to regulate Microcystis blooms in an eco-friendly manner.     

Relevance of the Diastereotopic Ligation of Magnesium Atoms of Chlorophylls in the Major Light-harvesting Complex II (LHC II) of Green Plants

The recent high-resolution crystal structure of LHC II [Liu et al. (2004) Nature 428: 287–292] makes possible an unprecedented insight into the stereochemical features of how chlorophylls (Chl)s are bound. The diastereotopic ligation generates four structurally different pigment types, two Chl a and two Chl b, which are distinguished not only by the groups in the 7-position (methyl in Chl a and formyl in Chl b) but also by the face of the tetrapyrrole to which the fifth magnesium ligand is bound. Within a LHC II monomer, out of the eight Chl a six have a ‚normal’ α-coordination and two are β-coordinated while out of the six Chl b only one has the ‚special’ β-coordination. In Photosystem I where a more meaningful statistical analysis could be made, out of 96 Chl a only 14 are β-coordinated, again indicating a preference for the ‚normal’ α-coordination [Balaban et al. (2002) Biochim Biophys Acta Bioenerget 1556: 197–207; Oba and Tamiaki (2002a) Photosynth Res 74: 1–10]. Astonishingly, all the special β-Chls are part of the stromal ring of Chls within the LHC II trimers and occupy key positions for the excitation energy transfer. Sequential energy traps are engineered with one hetero- and three homo-dimers. A careful pairing of carotenoids with the special β-Chls, which could quench their triplet states efficiently, implies a functional relevance of this diastereotopic ligation.     

Light-regulated pigment interconversion in pheophytin/chlorophyll-containing complexes formed during plant leaves greening

Upon illumination of etiolated maize leaves the photoconversion of protochlorophyllide Pchlide 655/650 into chlorophyllide Chlide 684/676 was observed. It was shown that chlorophyllide Chlide 684/676 in the dark is transformed into pheophytin Pheo 679/675 and chlorophyll Chl 671/668 by means of two parallel reactions, occurring at room temperature: Chlide 684/676. The formed pheophytin Pheo 679/675 was unstable and in the dark was transformed into chlorophyll Chl 671/668 in a few seconds: Pheo 679/675 Chl 671/668. The last reaction is reversed by the light: Chl/668 Pheo 679/675. Thus, on the whole in the greening etiolated leaves this process occurs according to the following scheme:The observed light-regulated interconversion of Mg-containing and Mg-free chlorophyll analogs is activated by ATP and inhibited by AMP.Abbreviations Chl- chlorophyll - Chlide- chlorophyllide - Pchlide- protochlorophyllide - Pheo- pheophytin - PS II RC- Photosystem II reaction centres. Abbreviations for native pigment forms: the first number after the pigment symbol corresponds to the maximum position of the low-temperature fluorescence band (nm), the second number to the maximum position of the longwave absorption band     

Peptide diversity in strains of the cyanobacterium <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> isolated from Portuguese water supplies

Strains of the cyanobacterium Microcystis aeruginosa were isolated into pure culture from a variety of lakes, rivers, and reservoirs in Portugal. Samples were tested with matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry (MALDI-TOF MS) to investigate the presence of various peptide groups including aeruginosins, microginins, anabaenopeptins, cyanopeptilins, microcystins, and microviridins and other peptide-like compounds. Binary data, based on the presence and absence of different peptide groups, were analyzed by phylogenetic inference. DNA was also extracted from the samples and tested using a range of primers. Those strains that gave positive results for a Microcystis-specific primer pair were further analyzed for the presence of genes linked to the biosynthesis of microginin and microcystin. The results showed that a wide range of microcystin forms were produced by the strains among which MC-LR, -FR, -RR, -WR, and -YR were the most common. The peptide profiles obtained from the MALDI analysis were assessed using cluster analysis which resulted in the formation of distinct groups or chemotypes.     

The Status and Characteristics of Eutrophication in the Yangtze River (Changjiang) Estuary and the Adjacent East China Sea, China

Eutrophication has become increasingly serious and noxious algal blooms have been of more frequent occurrence in the Yangtze River Estuary and in the adjacent East China Sea. In 2003 and 2004, four cruises were undertaken in three zones in the estuary and in the adjacent sea to investigate nitrate (NO₃–N), ammonium (NH₄–N), nitrite (NO₂–N), soluble reactive phosphorus (SRP), dissolved reactive silica (DRSi), dissolved oxygen (DO), phytoplankton chlorophyll a (Chl a) and suspended particulate matter (SPM). The highest concentrations of DIN (NO₃–N+NH₄–N+NO₂–N), SRP and DRSi were 131.6, 1.2 and 155.6 μM, respectively. The maximum Chl a concentration was 19.5 mg m⁻³ in spring. An analysis of historical and recent data revealed that in the last 40 years, nitrate and SRP concentrations increased from 11 to 97 μM and from 0.4 to 0.95 μM, respectively. From 1963 to 2004, N:P ratios also increased from 30–40 up to 150. In parallel with the N and P enrichment, a significant increase of Chl a was detected, Chl a maximum being 20 mg m⁻³, nearly four times higher than in the 1980s. In 2004, the mean DO concentration in bottom waters was 4.35 mg l⁻¹, much lower than in the 1980s. In comparison with other estuaries, the Yangtze River Estuary was characterized by high DIN and DRSi concentrations, with low SRP concentrations. Despite the higher nutrient concentrations, Chl a concentrations were lower in the inner estuary (Zones 1 and 2) than in the adjacent sea (Zone 3). Based on nutrient availability, SPM and hydrodynamics, we assumed that in Zones 1 and 2 phytoplankton growth was suppressed by high turbidity, large tidal amplitude and short residence time. Furthermore, in Zone 3 water stratification was also an important factor that resulted in a greater phytoplankton biomass and lower DO concentrations. Due to hydrodynamics and turbidity, the open sea was unexpectedly more sensitive to nutrient enrichment and related eutrophication processes.     

The phosphorus-chlorophyll a relationship in periphytic communities in a controlled ecosystem

The correlation between total dissolved phosphorus input concentrations and periphytic chlorophyll a concentrations was examined after a four-week colonization period on artificial substrates in large, continuous-flow microcosms. A significant correlation was established; the data produced a linear regression and substantiated a recently proposed curvilinear regression analysis. The curvilinear equation can be used in future microcosm studies as a predictive aid for regulating phosphorus input concentrations.     

Biomass-dependent effects of common carp on water quality in shallow ponds

We examined the biomass-dependent effects of common carp (Cyprinus carpio) on water quality in 10 ponds at the Eagle Mountain Fish Hatchery, Fort Worth, Texas, USA. Ponds contained 0–465 kg ha⁻¹ of common carp. We measured limnological variables at weekly intervals for four weeks in early summer, after which ponds were drained and the biomass of fish and macrophytes was determined. Common carp biomass was significantly positively correlated with chlorophyll a, total phosphorus, total nitrogen, and Keratella spp. density and negatively correlated to bushy pondweed (Najas guadalupensis) biomass. In addition, we combined our data with data from comparable studies to develop more robust regression models that predict the biomass-dependent effects of common carp on water quality variables across a wide range of systems.     

Water quality improvement and phytoplankton response in the drinking water source in Meiliang Bay of Lake Taihu, China

Water quality experienced changes throughout the 3-year ecological engineering experiment in the drinking water source in Meiliang Bay of Lake Taihu. Average concentrations of TN, TP, NH₄⁺, BOD₅ and transparency in the drinking water source during the period of July–December 2005 were 1.85, 0.13, 0.23, 3.03 mg L⁻¹ and 27.5 cm, respectively, decreasing by 47.9%, 21.2%, 83.3%, 54.4% and 24.2%, compared to concentrations from the same period in 2003. Concentrations of chlorophyll a and COD were 89.9 μg L⁻¹ and 6.45 mg L⁻¹, increasing by 27.9% and 17.7%, compared to the values in 2003. Cyanobacteria (mainly Microcystis) dominated the phytoplankton community in the ecological engineering area during July–December 2005. Densities of cyanobacteria and Microcystis were higher in 2005 than in 2004 and higher inside the engineering area than outside. Density percentages of cyanobacteria and Microcystis to total algae were above 90% and 60% during the bloom period. Average density of flagellate algae was higher during July–December 2005 than in 2004. Changes in water quality in the engineering area resulted mainly from the weakening of waves, decrease in concentrations of suspended solids, and assimilation of mass algae and periphytons. In spite of initial improvement of water quality, cyanobacterial bloom still determined the phytoplankton dynamics and variations. Additionally, nutrient concentration still remained at a high level without control of external loading. Therefore, a more holistic approach and long-term management should be adopted in Lake Taihu.     

Acute inhibition of protease and suppression of growth in zooplankter,Moina macrocopa,by Microcystis blooms collected in Central India

Cyanobacterial blooms consisting of Microcystis spp., collected from 14 water-bodies in Central India, and an adapted culture, were studied for likely impact on zooplankton community. When fed with single cells of Microcystis from several locations, in mixtures with Chlorella, population growth of the cladoceran Moina macrocopa was suppressed. Microcystis alone was unsuitable as food. In three cases, bloom extracts enhanced mortality of starved zooplankton. Extracts from several sources inhibited protease activity when trypsin or a crude extract from zooplankton served as enzyme source. Upon fractionation by solid-phase extraction, the C-18 passed extract contained the anti-protease and toxic substances for zooplankton, whereas a methanol eluted fraction retained the trypsin inhibitory substance. The study suggests that production of protease inhibitors by cyanobacteria is a factor responsible for feeding inhibition and mortality in zooplankton, which in turn could regulate the community structure of grazers.     

Application of luminescence for quality evaluation of in vitro regenerated strawberry plants

The parameters of chlorophyll a fluorescence induction were measured: F_v/F_m, S_c/F_m, Rf_d and coefficient of L_d delayed luminescence decay kinetics, related with a course of primary photosynthesis reactions on leaves of strawberry plants, cultured in vitro by means of the micropropagation methods. Strawberry plants cv. Ananasowa from in vitro cultures in optimal condition show significantly higher values of luminescence parameters indicating better condition of plants of this variety in comparison with the variety Senga Sengana. After temperature lowering, however, these values were more reduced than for plants of Senga Sengana, which can be interpreted as higher susceptibility of this variety to chill. Addition of BAP caused disturbance of primary photosynthesis reactions rate, particularly in lower temperature. Auxin 2,4-D had no effect on the luminescence parameters in comparison with control cultures. Dehydration stress strongly diminished the values of measured parameters for Ananasowa variety what indicates the inhibition of primary photosynthesis reaction in leaves. The old culture of Senga Sengana variety showed higher tolerance on linuron in comparison with the new one.     

Kryptoperidinium foliaceum blooms in South Carolina: a multi-analytical approach to identification

Observations following the discovery of Kryptoperidinium foliaceum blooms in South Carolina (SC), USA, suggest that a multi-analytical approach, using a standard, minimal set of criteria, should be adopted for determining dinoflagellate species identity and taxonomic placement. A combination of morphological, molecular, and biochemical analyses were used to determine the identity of this “red tide” dinoflagellate, first documented in SC waters in the spring of 1998. Results from thecal plate tabulations (based on scanning electron and epifluorescence microscopy), gene sequence data, species-specific PCR probe assays, and microalgal pigment profiles were analyzed and compared to reference cultures of K. foliaceum. Comparative data showed marked inconsistencies among the K. foliaceum reference culture isolates. In addition, the SC bloom isolate was shown to be mononucleate, contrary to previous reports for K. foliaceum, suggesting a more transient endosymbiotic association than previously considered.     

A note about Daphnia in Venezuelan waterbodies

After examining numerous water samples of 19 lakes and reservoirs in Venezuela, only two species of Daphnia were found: D. laevis (in colder, high altitude lakes) and D. gessneri. This observation supports the theory of the scarcity of Daphnia species in tropical inland waters.     

Biological and water quality effects of artificial mixing of Arbuckle Lake,Oklahoma, during 1977

This paper describes the effects of total lake mixing with 16 axial flow (Garton) pumps on the water quality, algal biomass and community metabolism of Arbuckle Lake, Oklahoma.Pumping began on July 1, 1977, and subsequently lowered the thermocline throughout the lake. The concentration of dissolved oxygen rose in formerly anoxic strata. Water quality in the former hypolimnion improved. Concentration of ammonia and BOD₅ decreased, and concentrations of manganese remained unchanged in 1977 compared to the control year (1976). But, concentrations of sulfide in the hypolimnion were higher in 1977 than in 1976. Algal biomass as chlorophyll a was about the same in 1977 as in 1978. The depth of the Secchi disc was also the same. An algal bloom did not occur. Pumping decreased the ratio gross production: community respiration as measured by a free water method, suggesting that lakes which are artificially mixed will have lower net primary productivities than lakes which are not artificially mixed.     

Products of Chlorophyll a Transformation by Selected Benthic Organisms in the Odra Estuary (Southern Baltic Sea)

The transformation of chlorophyll a by two benthic bivalves – Dreissena polymorpha and Mytilus edulis and, for comparison, by the chironomid Chironomus plumosus – was examined. D. polymorpha is a suspension-feeding, freshwater but very salinity-tolerant species, M. edulis a suspension-feeding marine species, and C.␣plumosus a typical detritivorous freshwater species. All three are common in the Odra Estuary. Specimens of the three species were collected from this area in the period 1999–2003. The bivalves were transferred to a compartment on land and kept in tanks filled with water (filtered and unfiltered) collected simultaneously with the organisms, under light and temperature conditions similar to those obtaining in their natural environment. Pigments in the water samples, faeces and pseudofaeces from the tanks, and in the gut content from C. plumosus, were determined with HPLC. The digestion products of the three species were mainly phaeophorbides a, generally thought to originate mainly from zooplankton grazing, and pyrophaeophytin a in minor quantities. The results indicate that the main chlorophyll a derivatives in faeces and pseudofaeces and gut content of the three species were the same as those in the sediments of the Baltic Sea, including the Odra Estuary. Our previous studies have shown that the sediments in this area are richer in phaeophorbides a than sediments from other parts of the southern Baltic Sea. All this suggests that the benthos may play an important role in the transformation of chlorophyll a in the Odra Estuary area.