Characterization of the state transition in the cyanobacterium synechococcus sp. 7002 and a phycobilisome-less Mutant

ABSTRACT Photosynthetic state transitions were investigated in the cyanobacterium Synechococcus sp. PCC 7002 in both wild-type cells and mutant cells lacking phycobilisomes. Preillumination in the presence of DCMU (3(3,4 dichlorophenyl) 1,1 dimethyl urea) induced state 1 and dark adaptation induced state 2 in both wild-type and mutant cells as determined by 77K fluorescence emission spectroscopy. Light-induced transitions were observed in the wildtype after preferential excitation of phycocyanin (state 2) or preferential excitation of chlorophyll .a. (state 1). The state 1 and 2 transitions in the wild-type had half-times of approximately 10 seconds. Cytochrome f and P-700 oxidation kinetics could not be correlated with any current state transition model as cells in state 1 showed faster oxidation kinetics regardless of excitation wavelength. Light-induced transitions were also observed in the phycobilisomeless mutant after preferential excitation of short wavelength chlorophyll !l. (state 2) or carotenoids and long wavelength chlorophyll it (state 1). One-dimensional electrophoresis revealed no significant differences in phosphorylation patterns of resolved proteins between wild-type cells in state 1 and state 2. It is concluded that the mechanism of the light state transition in cyanobacteria does not require the presence of the phycobilisome. The results contradict proposed models for the state transition which require an active role for the phycobilisome.

Pigment orientation changes detected by low temperature linear dichroism spectroscopy: cold-hardening transition in phycobilisome-containing organisms

Low temperature (77K) linear dichroism spectroscopy was used to characterize pigment orientation changes accompanying the light state transition in the cyanobacterium, Synechococcus sp. pee 6301, and cold-hardening in winter rye (Secale cereale L. cv. Puma). Samples were oriented for spectroscopy using the gel squeezing method (Abdourakhmanov et aI., 1979) and brought to 77K in liquid nitrogen. The linear dichroism (LD) spectra of Synechococcus 6301 phycobilisome/thylakoid membrane fragments cross-linked in light state 1 and light state 2 with glutaraldehyde showed differences in both chlorophyll a and phycobilin orientation. A decrease in the relative amplitude of the 681nm chlorophyll a positive LD peak was observed in membrane fragments in state 2. Reorientation of the phycobilisome (PBS) during the transition to state 2 resulted in an increase in core allophycocyanin absorption parallel to the membrane, and a decrease in rod phycocyanin parallel absorption. This result supports the "spillover" and "PBS detachment" models of the light state transition in PBS-containing organisms, but not the "mobile PBS" model. A model was proposed for PBS reorientation upon transition to state 2, consisting of a tilt in the antenna complex with respect to the membrane plane. Linear dichroism spectra of PBS/thylakoid fragments from the red alga, Porphyridium cruentum, grown in green light (containing relatively more PSI) and red light (containing relatively more PSll) were compared to identify chlorophyll a absorption bands associated with each photosystem. Spectra from red light - grown samples had a larger positive LD signal on the short wavelength side of the 686nm chlorophyll a peak than those from green light - grown fragments. These results support the identification of the difference in linear dichroism seen at 681nm in Synechococcus spectra as a reorientation of PSll chromophores. Linear dichroism spectra were taken of thylakoid membranes isolated from winter rye grown at 20°C (non-hardened) and 5°C (cold-hardened). Differences were seen in the orientation of chlorophyll b relative to chlorophyll a. An increase in parallel absorption was identified at the long-wavelength chlorophyll a absorption peak, along with a decrease in parallel absorption from chlorophyll b chromophores. The same changes in relative pigment orientation were seen in the LD of isolated hardened and non-hardened light-harvesting antenna complexes (LHCII). It was concluded that orientational differences in LHCII pigments were responsible for thylakoid LD differences. Changes in pigment orientation, along with differences observed in long-wavelength absorption and in the overall magnitude of LD in hardened and non-hardened complexes, could be explained by the higher LHCII monomer:oligomer ratio in hardened rye (Huner et ai., 1987) if differences in this ratio affect differential light scattering properties, or fluctuation of chromophore orientation in the isolated LHCII sample.

Biochemical and Biotechnological Studies on the Cyanobacterium Synechocystis salina Wislouch

The group cyanobacteria includes a large number of organisms characterised by a low state of cellular organization. Their cells lack a well defined nucleus. Cell division is by division of the protoplast by an ingrowth of the septum. These organisms are characterised generally by a blue green colouration of the cell, the chief pigments being chlorophyll-a, carotenes, xanthophylls, C phycocyanin and C phycoerythrin. The product of photosynthesis is glycogen. These organisms lack flagellate reproductive bodies and there is a total lack of sexual reproduction. They are also unique because of the presence of murein in the place of cellulose (cell wall) and the absence of chloroplast, mitochondria and endoplasmic reticulum. Just like bacteria some of them possess Plasmids and can fix atmospheric nitrogen. In the present study growth kinetics, heavy metal tolerance, tolerance mechanisms, heavy metal intake, and antibacterial activity of §ynechocystics salina Wislouch - a nanoplanktonic, euryhaline, Cyanobacterium present in Cochin back waters has been carried out for the potential biotechnological application of this organism. _§; salina occur as small spherical cells of 3n diameter (sometimes in pairs) with bluish green colour. The species is characterised by jerky movement of the cells and is structrually similar to other cyanobacteria

Deleterious effect of TRIS buffer on growth rates and pigment content of Gracilaria birdiae Plastino & EC Oliveira (Gracilariales, Rhodophyta)

This work evaluated the effects of Tris (hydroxymethyl)-aminomethane (TRIS) buffer and its interaction with nutrient concentration on the development of Gracilaria birdie, a common species on the Brazilian coast that has been exploited for agar production. Responses to different conditions were assessed through growth rates and pigment content (chlorophyll a, phycoerythrin, phycocyanin and allophycocyanin). Provasoli`s nutrient solution with and without TRIS addition was tested at concentrations of 12.5, 25 and 50%. The pH was also monitored. G. birdiae grew better in the absence of TRIS and at low nutrient concentrations, 12.5 and 25% (growth rates of 10.8-11.3%.day(-1)). Higher contents of phycoerythrin and chlorophyll a were observed without TRIS at 12.5 and 25% (Phycoerythrin, 649.6-698.0 mu g g(-1) fresh biomass; Chlorophyll a, 156.0-168.6 mu g g(-1) fresh biomass). These findings highlight the deleterious effect of TRIS on growth and phycoerythrin and chlorophyll a content. They also demonstrate the importance of appropriate nutrient concentration for laboratory cultures, depending on the intrinsic characteristics of each species.

Color inheritance and pigment characterization of red (wild-type), greenish-brown, and green strains of Gracilaria birdiae (Gracilariales, Rhodophyta)

Species of Gracilaria are some of the most useful algae in the world for the production of agar. As a consequence of its economic importance, the genus has been the subject of many studies worldwide. Color variants of Gracilaria birdiae have been found in the natural population on the Brazilian coast, and they have also been isolated from plants cultivated in laboratory. These findings raised new questions regarding intraspecific variation and the prospects of cultivating such variants for their agar production. Therefore, this work aimed to determine the mode of color inheritance for two G. birdiae strains: a greenish-brown strain (gb) found in a natural population and a green strain (gr) which had arisen as a spontaneous mutation in a red plant cultured in the laboratory. The pigment contents of these strains, as well as the red wildtype (rd), were also characterized. Crosses between female and male plants of the same color (rd, gr, or gb) and between different colors were performed. Crosses between plants of the same color showed tetrasporophytic and gametophytic descendents of the parental color. Recessive nuclear inheritance was found in the greenish-brown strain, and cytoplasmic maternal inheritance was found in the green strain; both had lower phycoerythrin and higher concentrations of allophycocyanin and phycocyanin than the wild-type. Chlorophyll a contents were similar among all strains. Taken together, our results contribute to knowledge about the variability of this important red algae. In addition, since greenish-brown and green strains showed stability of color, both could be selected and tested in experimental sea cultivation to evaluate if mutants have advantageous performance when compared with red strain.

Effects of kinetin and nitrogen on growth rates, pigment and protein contents in wild and phycoerythrin-deficient strains of Hypnea musciformis (Rhodophyta)

Hypnea musciformis (Wulfen in Jacqu.) J.V. Lamour. is the main source for carrageenan production in Brazil and strains with selected characteristics could improve the production of raw material. The effects of kinetin on growth rates, morphology, protein content, and concentrations of pigments (chlorophyll a, phycoerythrin, phycocyanin, and allophycocyanin) were assessed in the wild strain (brown phenotype) and in the phycoerythrin-deficient strain (green phenotype) of H. musciformis. Concentrations of kinetin ranging from 0 to 50 mu M were tested in ASP 12-NTA synthetic medium with 10 mu M nitrate (N-limited) and 100 mu M nitrate (N-saturated). In N-limited condition, kinetin stimulated growth rates of the phycoerythrin-deficient strain and formation of lateral branches in both colour strains. Kinetin stimulated protein biosynthesis in both strains. However, differences between both nitrogen conditions were significant only in the phycoerythrin-deficient strain. In the wild strain, effects of kinetin on concentrations of phycobiliproteins were not significant in both nitrogen conditions, except for chlorophyll content. However, the phycoerythrin-deficient strain showed an opposite response, and kinetin stimulated the phycobiliprotein biosynthesis, with the highest concentrations of phycoerythrin in N-saturated medium, while the highest concentrations of allophycocyanin and phycocyanin were observed in N-limited medium. These results indicate that the effects of kinetin on growth, morphology, protein and phycobiliprotein contents are influenced by nitrogen availability, and the main nitrogen storage pools in phycoerythrin-deficient strain of H. musciformis submitted to N-limited conditions were phycocyanin and allophycocianin, the biosynthesis of which was enhanced by kinetin.

Growth responses and photosynthetic characteristics of wild and phycoerythrin-deficient strains of Hypnea musciformis (Rhodophyta)

The phycoerythrin-deficient strain (green phenotype) of Hypnea musciformis (Rhodophyta) originated from a green branch, which had arisen as a spontaneous mutation in a wild plant (brown phenotype) collected from the Brazilian coast. The present study describes the growth responses to irradiance, photoperiod and temperature variations, pigment contents, and photosynthetic characteristics of the brown and green strains of H. musciformis. The results showed that growth rates increased as a function of irradiance (up to 40 mu mol photons m(-2) s(-1)) but, with further increase in irradiance (from 40 to 120 mu mol photons m(-2) s(-1)), became light-saturated and remained almost unchanged. The highest growth rates of the brown and green strains were observed in temperatures of 20-25 degrees C under long (14:10 h LD) and short (10:14 h LD) photoperiods. The brown strain had higher growth rates than the green strain in the short photoperiod, which could be related to the high concentrations of phycobiliproteins. Phycoerythrin was not detected in the green strain. The brown strain had higher concentrations of allophycocyanin and phycoerythrin in the short photoperiod while the green strain had higher concentrations of phycocyanin. The brown strain presented higher photosynthetic efficiency (alpha), and lower saturation parameter (I-k) and compensation irradiance (I-c) than the green strain. The brown strain exhibited the characteristics of shade-adapted plants, and its higher value of photosynthetic efficiency could be attributed to the higher phycoerythrin concentrations. Results of the present study indicate that both colour strains of H. musciformis could be selected for aquaculture, since growth rates were similar (although in different optimal light conditions), as the green strain seems to be adapted to higher light levels than the brown strain. Furthermore, these colour strains could be a useful experimental system to understand the regulation of biochemical processes of photosynthesis and metabolism of light-harvesting pigments in red algae.

Effects of temperature, irradiance and photoperiod on growth and pigment content in some freshwater red algae in culture

The responses of relative growth rate (% day-1) and pigment content (chlorophyll a, phycocyanin and phycoerythrin) to temperature, irradiance and photoperiod were analyzed in culture in seven freshwater red algae: Audouinella hermannii (Roth) Duby, Audouinella pygmaea (Kützing) Weber-van Bosse, Batrachospermum ambiguum Montagne, Batrachospermum delicatulum (Skuja) Necchi et Entwisle, 'Chantransia' stages of B. delicatulum and Batrachospermum macrosporum Montagne and Compsopogon coeruleus (C. Agardh) Montagne. Experimental conditions included temperatures of 10, 15, 20 and 25°C and low and high irradiances (65 and 300 μmol photons m-2 s-1, respectively). Long and short day lengths (16:8 and 8:16 LD cycles) were also applied at the two irradiances. Growth effects of temperature and irradiance were evident in most algae tested, and there were significant interactions among treatments. Most freshwater red algae had the best growth under low irradiance, confirming the preference of freshwater red algae for low light regimens. In general there was highest growth rate in long days and low irradiance. Growth optima in relation to temperature were species-specific and also varied between low and high irradiances for the same alga. The most significant differences in pigment content were related to temperature, whereas few significant differences could be attributed to variation in irradiance and photoperiod or interactions among the three parameters. The responses were species-specific and also differed for pigments in distinct temperatures, irradiances and photoperiods in the same alga. Phycocyanin was generally more concentrated than phycoerythrin and phycobiliproteins were more concentrated than chlorophyll a. The highest total pigment contents were found in two species typical of shaded habitats: A. hermannii and C. coeruleus. The expected inverse relationship of pigment with irradiance was observed only in C. coeruleus. In general, the most favorable conditions for growth were not coincident with those with highest pigment contents.

Ecology and morphological characterization of gametophyte and 'Chantransia' stages of Sirodotia huillensis (Batrachospermales, Rhodophyta) from a stream in central Mexico

The morphology and phenology of Sirodotia huillensis was evaluated seasonally in a central Mexican first-order calcareous stream. Water temperature was constant (24-25°C) and pH circumneutral to alkaline (6.7-7.9), and calcium and sulfates were the dominant ions. The gametophyte stages were characterized by the presence of a distinctive mucilaginous layer, a marked difference in phycocyanin to phycoerythrin ratio between female and male plants, and the presence of a carpogonia with a large trichogyne (>60 μm). Occasionally three capogonia were observed on a single basal cell. The 'Chantransia' stages were morphologically similar to those described for the other members of Batrachospermales. A remarkable observation was the formation of dome-shaped structures, consisting of prostrate filaments that are related with the development of new gametophytes. Chromosome numbers were n = 4 for fascicle cells, cortical filament cells and dome-shaped cells, and 2n = 8 for gonimoblast filament cells and 'Chantransia' stage filaments. Gametophytes and 'Chantransia' stages occurred in fast current velocities (60-170 cm/s) and shaded (33.1-121 μmol photons/m2/s) stream segments. The population fluctuated throughout the study period in terms of percentage cover and frequency: the 'Chantransia' stages were most abundant in the rainy season, whereas gametophytic plants had the highest frequency values during the dry season. These results were most likely a result of fluctuations in rainfall and related changes in current velocity. Some characteristics of this population can be viewed as probable adaptations to high current velocities: the mucilaginous layer around plants that reduces drag; potential increase in fertilization by the elongate and plentiful trichogynes and abundant dome-shaped structures producing several gametophytes.

Sustainable production of biologically active molecules of marine based origin

The marine environment offers both economic and scientific potential which are relatively untapped from a biotechnological point of view. These environments whilst harsh are ironically fragile and dependent on a harmonious life form balance. Exploitation of natural resources by exhaustive wild harvesting has obvious negative environmental consequences. From a European industry perspective marine organisms are a largely underutilised resource. This is not due to lack of interest but due to a lack of choice the industry faces for cost competitive, sustainable and environmentally conscientious product alternatives. Knowledge of the biotechnological potential of marine organisms together with the development of sustainable systems for their cultivation, processing and utilisation are essential. In 2010, the European Commission recognised this need and funded a collaborative RTD/SME project under the Framework 7-Knowledge Based Bio-Economy (KBBE) Theme 2 Programme 'Sustainable culture of marine microorganisms, algae and/or invertebrates for high value added products'. The scope of that project entitled 'Sustainable Production of Biologically Active Molecules of Marine Based Origin' (BAMMBO) is outlined. Although the Union is a global leader in many technologies, it faces increasing competition from traditional rivals and emerging economies alike and must therefore improve its innovation performance. For this reason innovation is placed at the heart of a European Horizon 2020 Strategy wherein the challenge is to connect economic performance to eco performance. This article provides a synopsis of the research activities of the BAMMBO project as they fit within the wider scope of sustainable environmentally conscientious marine resource exploitation for high-value biomolecules. © 2013 Elsevier B.V.

Filogenia molecular de cianobactérias baseada em sequências do 16S-23S-ITS rDNA e PC-IGS: investigação de transferência lateral do PC

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detecção molecular quantitativa de cianobactérias hepatotóxicas através de PCR competitiva

Pós-graduação em Ciências Biológicas (Biologia Vegetal) - IBRC

Inferência espacial de cianobactérias a partir da fluorescência in vivo do pigmento ficocianina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da ocorrência de cianobactérias no reservatório de Nova Avanhandava-SP por meio da inferência do pigmento ficocianina

Pós-graduação em Ciências Cartográficas - FCT

Cyanobacteria detection in Guarapiranga Reservoir (São Paulo State, Brazil) using Landsat TM and ETM+ images

Algae bloom is one of the major consequences of the eutrophication of aquatic systems, including algae capable of producing toxic substances. Among these are several species of cyanobacteria, also known as blue-green algae, that have the capacity to adapt themselves to changes in the water column. Thus, the horizontal distribution of cyanobacteria harmful algae blooms (CHABs) is essential, not only to the environment, but also for public health. The use of remote sensing techniques for mapping CHABs has been explored by means of bio-optical modeling of phycocyanin (PC), a unique inland waters cyanobacteria pigment. However, due to the small number of sensors with a spectral band of the PC absorption feature, it is difficult to develop semi-analytical models. This study evaluated the use of an empirical model to identify CHABs using TM and ETM+ sensors aboard Landsat 5 and 7 satellites. Five images were acquired for applying the model. Besides the images, data was also collected in the Guarapiranga Reservoir, in São Paulo Metropolitan Region, regarding the cyanobacteria cell count (cells/mL), which was used as an indicator of CHABs biomass. When model values were analyzed excluding calibration factors for temperate lakes, they showed a medium correlation (R²=0.81, p=0.036), while when the factors were included the model showed a high correlation (R²=0.96, p=0.003) to the cyanobacteria cell count. The empirical model analyzed proved useful as an important tool for policy makers, since it provided information regarding the horizontal distribution of CHABs which could not be acquired from traditional monitoring techniques.