RESUMEN
Cyanobacteria stand out among phytoplankton when they form massive blooms and produce toxins. Because cyanotoxin genes date to the origin of metazoans, the hypothesis that cyanotoxins function as a defense against herbivory is still debated. Although their primary cellular function might vary, these metabolites could have evolved as an anti-predator response. Here we evaluated the physiological and molecular responses of a saxitoxin-producing Raphidiopsis raciborskii to infochemicals released by the grazer Daphnia gessneri. Induced chemical defenses were evidenced in R. raciborskii as a significant increase in the transcription level of sxt genes, followed by an increase in saxitoxin content when exposed to predator cues. Moreover, cyanobacterial growth decreased, and no significant effects on photosynthesis or morphology were observed. Overall, the induced defense response was accompanied by a trade-off between toxin production and growth. These results shed light on the mechanisms underlying zooplankton-cyanobacteria interactions in aquatic food webs. The widespread occurrence of the cyanobacterium R. raciborskii in freshwater bodies has been attributed to its phenotypic plasticity. Assessing the potential of this species to thrive over interaction filters such as zooplankton grazing pressure can enhance our understanding of its adaptive success.
Asunto(s)
Cylindrospermopsis , Daphnia/metabolismo , Feromonas/metabolismo , Saxitoxina , Zooplancton/metabolismo , Animales , Cylindrospermopsis/genética , Cylindrospermopsis/crecimiento & desarrollo , Cylindrospermopsis/metabolismo , Cadena Alimentaria , Regulación Bacteriana de la Expresión Génica , Genes Bacterianos , Saxitoxina/biosíntesis , Saxitoxina/genéticaRESUMEN
Cylindrospermopsis raciborskii is a potentially toxic freshwater cyanobacterium that can tolerate a wide range of light and temperature. Due to climatic changes, the interaction between light and temperature is studied in aquatic systems, but no study has addressed the effect of both variables on the saxitoxins production. This study evaluated the combined effect of light and temperature on saxitoxins production and cellular quota in C. raciborskii. Experiments were performed with three C. raciborskii strains in batch cultures under six light intensities (10, 40, 60, 100, 150, and 500 µmol of photons m-2 s-1) and four temperatures (15, 20, 25, and 30 °C). The growth of C. raciborskii strains was limited at lower temperatures and the maximum growth rates were obtained under higher light combined with temperatures equal or above 20 °C, depending on the strain. In general, growth was highest at 30 °C at the lower light intensities and equally high at 25 °C and 30 °C under higher light. Highest saxitoxins concentration and cell-quota occurred at 25 °C under high light intensities, but were much lower at 30 °C. Hence, increased temperatures combined with sufficient light will lead to higher C. raciborskii biomass, but blooms could become less toxic in tropical regions.
Asunto(s)
Cylindrospermopsis , Luz , Saxitoxina/metabolismo , Temperatura , Cylindrospermopsis/crecimiento & desarrollo , Cylindrospermopsis/metabolismo , Cylindrospermopsis/efectos de la radiaciónRESUMEN
Cyanobacterial blooms are characterized by intense growth of one or few species that will dominate the phytoplankton community for periods of few months to an entire year or more. However, even during persistent blooms, important seasonal changes among dominant species can be observed. Pampulha reservoir is a tropical eutrophic reservoir presenting permanent blooms. To identify the main species in this environment, a closer analysis performed by microscopy and 16S-rRNA DGGE revealed Cylindrospermopsis raciborskii as highly dominant throughout the year. The second most abundant group comprised species belonging to the Microcystis genus. They followed a well-defined seasonal pattern described by interesting species-specific ecological trends. During thermal stratification in the rainy/warmer season, C. raciborskii dominated in the water column, while Microcystis spp. were abundant at the end of the dry season, a period characterized by higher total phosphorus concentrations. Phylogenetic analyses confirmed the two dominant taxa and their seasonal trends. The results showed that cyanobacteria major controlling factors were strongly species dependent, shifting from physical/climate related (stratification) to more chemical driven (nutrients/eutrophication). Identifying these drivers is therefore essential for the understanding of the bloom dynamics and the real risks associated with each species, and to eventually adopt the most appropriate and effective management strategies.
Asunto(s)
Cylindrospermopsis/clasificación , Cylindrospermopsis/crecimiento & desarrollo , Eutrofización/fisiología , Microcystis/clasificación , Microcystis/crecimiento & desarrollo , Cylindrospermopsis/genética , Microcystis/genética , Fósforo/análisis , Filogenia , Fitoplancton/clasificación , ARN Ribosómico 16S/genética , Estaciones del AñoRESUMEN
In tropical and subtropical lakes, eutrophication often leads to nuisance blooms of Cylindrospermopsis raciborskii. In laboratory experiments, we tested the combined effects of flocculant polyaluminum chloride (PAC) and lanthanum-modified bentonite (LMB) on the sinking and growth rates of three C. raciborskii strains. We tested the hypothesis that the combination of PAC and LMB would (1) effectively sink C. raciborskii in a test tube experiment and (2) impair C. raciborskii growth, irrespective of the biomass of the inoculum (bloom) and the strain in the growth experiment. We tested the recommended (LMB1) and a three-times higher dose of LMB (LMB3). The combined addition of PAC and LMB enhanced the sedimentation of all C. raciborskii strains. Moreover, both the PAC and LMB doses decreased the phosphate concentration. PAC and LMB1 decreased the growth rate of all strains, but the efficacy depended on the biomass and strain. The combined addition of PAC and LMB3 inhibited the growth of all strains independently of the biomass and strain. We conclude that a low dose of PAC in combination with the recommended dose of LMB decreases C. raciborskii blooms and that the efficiency of the technique depends on the biomass of the bloom. A higher dose of LMB is needed to obtain a more efficient control of C. raciborskii blooms.
Asunto(s)
Hidróxido de Aluminio , Bentonita , Cylindrospermopsis/crecimiento & desarrollo , Lantano , Bentonita/química , Biomasa , Clorofila/metabolismo , Clorofila A , Cylindrospermopsis/metabolismo , Lantano/química , Fósforo/metabolismoRESUMEN
Cylindrospermopsis raciborskii CS-505 is an invasive freshwater filamentous cyanobacterium that when grown diazotrophically may develop trichomes of up to 100 vegetative cells while differentiating only two end heterocysts, the sole sites for their N2-fixation process. We examined the diazotrophic growth and intercellular transfer mechanisms in C. raciborskii CS-505. Subjecting cultures to a combined-nitrogen-free medium to elicit N2 fixation, the trichome length remained unaffected while growth rates decreased. The structures and proteins for intercellular communication showed that while a continuous periplasmic space was apparent along the trichomes, the putative septal junction sepJ gene is divided into two open reading frames and lacks several transmembrane domains unlike the situation in Anabaena, differentiating a 5-fold higher frequency of heterocysts. FRAP analyses also showed that the dyes calcein and 5-CFDA were taken up by heterocysts and vegetative cells, and that the transfer from heterocysts and 'terminal' vegetative cells showed considerably higher transfer rates than that from vegetative cells located in the middle of the trichomes. The data suggest that C. raciborskii CS-505 compensates its low-frequency heterocyst phenotype by a highly efficient transfer of the fixed nitrogen towards cells in distal parts of the trichomes (growing rapidly) while cells in central parts suffers (slow growth).
Asunto(s)
Cylindrospermopsis/fisiología , Fijación del Nitrógeno , Tricomas/crecimiento & desarrollo , Anabaena/genética , Anabaena/fisiología , Proteínas Bacterianas/metabolismo , Biología Computacional , Cylindrospermopsis/genética , Cylindrospermopsis/crecimiento & desarrollo , Cylindrospermopsis/ultraestructura , Fluoresceínas/metabolismo , Regulación Bacteriana de la Expresión Génica , Microscopía Electrónica de Transmisión , Nitrogenasa/metabolismo , Sistemas de Lectura Abierta , Periplasma/metabolismo , Fenotipo , Tricomas/fisiologíaRESUMEN
Different environmental nitrogen sources play selective roles in the development of cyanobacterial blooms and noxious effects are often exacerbated when toxic cyanobacteria are dominant. Cylindrospermopsis raciborskii CS-505 (heterocystous, nitrogen fixing) and Raphidiopsis brookii D9 (non-N2 fixing) produce the nitrogenous toxins cylindrospermopsin (CYN) and paralytic shellfish toxins (PSTs), respectively. These toxin groups are biosynthesized constitutively by two independent putative gene clusters, whose flanking genes are target for nitrogen (N) regulation. It is not yet known how or if toxin biosynthetic genes are regulated, particularly by N-source dependency. Here we show that binding boxes for NtcA, the master regulator of N metabolism, are located within both gene clusters as potential regulators of toxin biosynthesis. Quantification of intra- and extracellular toxin content in cultures at early stages of growth under nitrate, ammonium, urea and N-free media showed that N-sources influence neither CYN nor PST production. However, CYN and PST profiles were altered under N-free medium resulting in a decrease in the predicted precursor toxins (doCYN and STX, respectively). Reduced STX amounts were also observed under growth in ammonium. Quantification of toxin biosynthesis and transport gene transcripts revealed a constitutive transcription under all tested N-sources. Our data support the hypothesis that PSTs and CYN are constitutive metabolites whose biosynthesis is correlated to cyanobacterial growth rather than directly to specific environmental conditions. Overall, the constant biosynthesis of toxins and expression of the putative toxin-biosynthesis genes supports the usage of qPCR probes in water quality monitoring of toxic cyanobacteria.
Asunto(s)
Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/biosíntesis , Cianobacterias/metabolismo , Cylindrospermopsis/metabolismo , Regulación Bacteriana de la Expresión Génica , Fijación del Nitrógeno , Venenos/metabolismo , Alcaloides , Compuestos de Amonio/metabolismo , Proteínas Bacterianas/genética , Toxinas Bacterianas/química , Técnicas de Cultivo Celular por Lotes , Brasil , Cianobacterias/crecimiento & desarrollo , Cianobacterias/aislamiento & purificación , Toxinas de Cianobacterias , Cylindrospermopsis/crecimiento & desarrollo , Cylindrospermopsis/aislamiento & purificación , Agua Dulce/microbiología , Estructura Molecular , Nitratos/metabolismo , Venenos/química , Queensland , Uracilo/análogos & derivados , Uracilo/biosíntesis , Uracilo/química , Urea/metabolismoRESUMEN
Cylindrospermopsis raciborskii is the focus of many studies due to its toxicity and increasing blooms frequency. The comprehension of the factors that might trigger these blooms is fundamental for the maintenance of good freshwater quality. To better understand the autoecology of C. raciborskii and to identify the factors controlling its dominance, general limnological features were evaluated in Pedalinhos reservoir, Brazil. Samplings were performed monthly between 2011 and 2013. Although C. raciborskii is considered a relatively perennial species in the tropics, it presented an interesting nonunimodal variation, with biovolume varying from 0 to c. 30 mm³ L⻹ in short time intervals (< 30 days). These temporal trends allowed the evaluation of the isolated effects of nonclimatic variables. Cylindrospermopsis raciborskii was connected with several variables, but nitrogen was the major determinant of its dynamics. Blooms were observed when dissolved inorganic nitrogen (DIN) was scarce (< 50 µg L⻹), mainly in relation to the availability of phosphorus and total-N (DIN : total-P < 3 and DIN : total-N < 0.04). This paper contributes to establish strategies to avoid C. raciborskii blooms, suggesting that a management in P levels would be not sufficient. It is necessary to consider N availability to avoid its dominance in the reservoir.
Asunto(s)
Cylindrospermopsis/crecimiento & desarrollo , Eutrofización , Agua Dulce/química , Nitrógeno/química , Brasil , Ambiente , Fósforo/química , Estaciones del Año , Calidad del Agua , Abastecimiento de AguaRESUMEN
The cosmopolitan and increasing distribution of Cylindrospermopsis raciborskii can be attributed to its ecophysiological plasticity and tolerance to changing environmental factors in water bodies. In reservoirs in the semi-arid region of Brazil, the presence and common dominance of C. raciborskii have been described in waters that are considered hard. We investigated the response of a Brazilian C. raciborskii strain to water hardness by evaluating its growth and saxitoxin production. Based on environmental data, a concentration of 5 mM of different carbonate salts was tested. These conditions affected growth either positively (MgCO3) or negatively (CaCO3 and Na2CO3). As a control for the addition of cations, MgCl2, CaCl2 and NaCl were tested at 5 or 10 mM, and MgCl2 stimulated growth, NaCl slowed but sustained growth, and CaCl2 inhibited growth. Most of the tested treatments increased the saxitoxin (STX) cell quota after six days of exposure. After 12 days, STX production returned to concentrations similar to that of the control, indicating an adaptation to the altered water conditions. In the short term, cell exposure to most of the tested conditions favored STX production over neoSTX production. These results support the noted plasticity of C. raciborskii and highlight its potential to thrive in hard waters. Additionally, the observed relationship between saxitoxin production and water ion concentrations characteristic of the natural environments can be important for understanding toxin content variation in other harmful algae that produce STX.
Asunto(s)
Cylindrospermopsis/metabolismo , Saxitoxina/biosíntesis , Agua/química , Brasil , Cloruro de Calcio/química , Cylindrospermopsis/crecimiento & desarrollo , Cloruro de Magnesio/química , Cloruro de Sodio/química , Factores de TiempoRESUMEN
Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju (Ordem Nostocales) is one of the most troublesome bloom-forming species in Brazil. Understanding the population dynamics of the different morphotypes of C. raciborskii (straight and coiled) could assist in the prediction of favourable conditions for the proliferation of this potentially toxin-producing species. The aim of the present study was to assess the effects of two different light intensities and temperatures on the growth rate and morphology of the trichomes of the straight and coiled morphotypes. For such, two non-toxin producing strains of C. raciborskii were used - one with a coiled trichome (ITEP31) and another with a straight trichome (ITEP28). The strains were cultured in BG-11 medium in a climatic chamber under controlled conditions. Two light intensities (30 and 90 µmol.m-2.s-1 ) were combined at temperatures of 21 and 31 °C and the growth rate and morphological changes were analysed. The morphotypes responded differently to the different temperatures and light intensities. Both strains exhibited faster growth velocities when submitted to higher light intensity and temperature. The lower temperature and higher luminosity hampered the development of both strains. Variations in cellular morphology and an absence of akinetes in both strains were related to the lower temperature (21 °C). The coiled morphotype demonstrated considerable phenotype plasticity, changing the morphology of trichome throughout its growth curve. Although molecular analysis does not sustain the separation of the morphotypes as distinct species, their different eco-physiological responses should be considered further knowledge of extreme importance for the population control of these potentially toxic organisms.
Asunto(s)
Cylindrospermopsis/crecimiento & desarrollo , Luz , Temperatura , Cylindrospermopsis/citología , Agua Dulce/microbiologíaRESUMEN
Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju (Ordem Nostocales) is one of the most troublesome bloom-forming species in Brazil. Understanding the population dynamics of the different morphotypes of C. raciborskii (straight and coiled) could assist in the prediction of favourable conditions for the proliferation of this potentially toxin-producing species. The aim of the present study was to assess the effects of two different light intensities and temperatures on the growth rate and morphology of the trichomes of the straight and coiled morphotypes. For such, two non-toxin producing strains of C. raciborskii were used - one with a coiled trichome (ITEP31) and another with a straight trichome (ITEP28). The strains were cultured in BG-11 medium in a climatic chamber under controlled conditions. Two light intensities (30 and 90 µmol.m-2.s-1 ) were combined at temperatures of 21 and 31 °C and the growth rate and morphological changes were analysed. The morphotypes responded differently to the different temperatures and light intensities. Both strains exhibited faster growth velocities when submitted to higher light intensity and temperature. The lower temperature and higher luminosity hampered the development of both strains. Variations in cellular morphology and an absence of akinetes in both strains were related to the lower temperature (21 °C). The coiled morphotype demonstrated considerable phenotype plasticity, changing the morphology of trichome throughout its growth curve. Although molecular analysis does not sustain the separation of the morphotypes as distinct species, their different eco-physiological responses should be considered further knowledge of extreme importance for the population control of these potentially toxic organisms.
Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju (Ordem Nostocales) é uma das espécies formadoras de florações mais prejudiciais no Brasil. Entender a dinâmica populacional dos diferentes morfotipos de C. raciborskii (reto e espiralado) pode auxiliar a previsão de condições favoráveis à proliferação e ao estabelecimento dessa espécie potencialmente produtora de saxitoxinas e cilindrospermopsina. O objetivo deste estudo foi avaliar os efeitos de duas diferentes intensidades luminosas e temperaturas na velocidade de crescimento e na morfologia dos tricomas de morfotipos reto e espiralado. Para tanto, foram utilizadas duas linhagens de C. raciborskii, não produtoras de toxinas, sendo uma com tricoma espiralado (ITEP31) e outra com tricoma reto (ITEP28), cultivadas em câmaras climáticas sob condições controladas em meio BG-11. Duas intensidades luminosas, 30 e 90 µmol.m-2.s-1 , foram combinadas com temperaturas de 21 e 31 °C e avaliadas as velocidades de crescimento e modificações morfológicas. Os morfotipos responderam de maneiras diferentes às temperaturas e intensidades luminosas. As linhagens investigadas, independentemente do morfotipo, apresentaram maiores velocidades de crescimento em intensidade luminosa e temperatura altas. A condição de temperatura baixa e intensidade luminosa alta prejudicaram o desenvolvimento de ambas as linhagens. Variações na morfometria celular, bem como a ausência de acinetos para ambas as linhagens, estiveram relacionadas às condições de temperatura baixa (21 °C). O morfotipo espiralado apresentou grande plasticidade fenotípica, alterando a morfologia do tricoma ao longo de sua curva de crescimento. Apesar de a análise com ferramentas moleculares não sustentar a separação dos morfotipos em espécies distintas, devem-se considerar as diferentes respostas ecofisiológicas entre os mesmos como conhecimento de extrema importância no controle populacional desses organismos potencialmente tóxicos.
Asunto(s)
Cylindrospermopsis/crecimiento & desarrollo , Luz , Temperatura , Cylindrospermopsis/citología , Agua Dulce/microbiologíaRESUMEN
The cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii are bloom-forming species common in eutrophic freshwaters. These filamentous species share certain physiological traits which imply that they might flourish under similar environmental conditions. We compared the distribution of the two species in a large database (940 samples) covering different climatic regions and the Northern and Southern hemispheres, and carried out laboratory experiments to compare their morphological and physiological responses. The environmental ranges of the two species overlapped with respect to temperature, light and total phosphorus (TP); however, they responded differently to environmental gradients; C. raciborskii biovolume changed gradually while P. agardhii shifted sharply from being highly dominated to a rare component of the phytoplankton. As expected, P. agardhii dominates the phytoplankton with high TP and low light availability conditions. Contrary to predictions, C. raciborskii succeeded in all climates and at temperatures as low as 11 °C. Cylindrospermopsis raciborskii had higher phenotypic plasticity than P. agardhii in terms of pigments, individual size and growth rates. We conclude that the phenotypic plasticity of C. raciborskii could explain its ongoing expansion to temperate latitudes and suggest its future predominance under predicted climate-change scenarios.
Asunto(s)
Cianobacterias/crecimiento & desarrollo , Agua Dulce/microbiología , Clima , Cianobacterias/fisiología , Cylindrospermopsis/crecimiento & desarrollo , Cylindrospermopsis/fisiología , Ambiente , Eutrofización , Fitoplancton/crecimiento & desarrollo , Fitoplancton/fisiología , TemperaturaRESUMEN
Cylindrospermopsis raciborskii (Woloszynska) Seenaya & Subba Raju is a freshwater cyanobacterium of worldwide distribution. In the North-eastern region of Brazil many eutrophic water reservoirs are characterized by the dominance of C. raciborskii, with recurrent occurrence of blooms. These water bodies have high conductivity due to a high ionic concentration, and are defined as hard (with high values of CaCO3). In this study, we investigated the long-term effect (12 days) of high calcium concentration (8 mM Ca2+) on C. raciborskii (T3 strain) growth, morphology, toxin content, and metabolism. Changes in protein expression profiles were investigated by proteomic analysis using 2D gel electrophoresis and mass spectrometry. A continued exposure to calcium had a pronounced effect on C. raciborskii (T3): it limited growth, decreased thricome length, increased chlorophyll-a content, altered toxin profile (although did not affect PST content, saxitoxin + neosaxitoxin), and inhibited the expression of proteins related to primary metabolism.