RESUMO
Samarium (Sm) is a rare-earth element recently included in the list of critical elements due to its vital role in emerging new technologies. With an increasing demand for Sm, microbial bioremediation may provide a cost-effective and a more ecologically responsible alternative to remove and recover Sm. We capitalized on a previously selected Chlamydomonas reinhardtii strain tolerant to Sm (1.33 × 10-4 M) and acidic pH and carried out settling selection to increase the Sm uptake performance. We observed a rapid response to selection in terms of cellular phenotype. Cellular size decreased and circularity increased in a stepwise manner with every cycle of selection. After four cycles of selection, the derived CSm4 strain was significantly smaller and was capable of sequestrating 41% more Sm per cell (1.7 × 10-05 ± 1.7 × 10-06 ng) and twice as much Sm in terms of wet biomass (4.0 ± 0.4 mg Sm · g-1) compared to the ancestral candidate strain. The majority (~70%) of the Sm was bioaccumulated intracellularly, near acidocalcisomes or autophagic vacuoles as per TEM-EDX microanalyses. However, Sm analyses suggest a stronger response toward bioabsorption resulting from settling selection. Despite working with Sm and pH-tolerant strains, we observed an effect on fitness and photosynthesis inhibition when the strains were grown with Sm. Our results clearly show that phenotypic selection, such as settling selection, can significantly enhance Sm uptake. Laboratory selection of microalgae for rare-earth metal bioaccumulation and sorption can be a promising biotechnological approach.
Assuntos
Chlamydomonas reinhardtii , Chlamydomonas reinhardtii/metabolismo , Chlamydomonas reinhardtii/genética , Biodegradação Ambiental , Seleção GenéticaRESUMO
Rare Earth Elements (REE) increasing demand prompts the research of biotechnological approaches to exploit secondary resources. We made use of the adapted Fluctuation analyses experiment to obtain Chlamydomonas reinhardtii ChlA strains resistant to Samarium (Sm) as the reference REE. The starting hypothesis was that adaptation to metal-containing media leads to an enhanced metal uptake. ChlA was able to adapt to 1.33·10-4 Sm M and pH~3 by pre-existing genetic variability, allowing the evolutionary rescue of 13 of the 99 populations studied. The rescuing resistant genotypes presented a mutation rate of 8.65·10-7 resistant cells per division. The resulting resistant population contradicted the expected fitness cost associated with the adaptation to Sm, selection resulted in larger and faster-growing resistant cells. Among the three isolated strains studied for Sm uptake, only one presented uplifted performance compared to the control population (46.64 µg Sm g-¹ of wet biomass and 3.26·10-7 ng Sm per cell, mainly bioaccumulated within the cells). The selection of microalgae strains with improved tolerance to REEs by this methodology could be a promising solution for REES sequestration. However, increased tolerance can be independent or have negative effects on uptake performance and cellular features studied are not directly correlated with the metal uptake. SUMMARY SENTENCE: Repurposing a classic laboratory evolution experiment to select for microalgae Samarium adapted strains for metals recovery and biotechnology approaches. DATA AVAILABILITY STATEMENT: All data generated or analyzed during this study are included in this published article (and its raw files).
Assuntos
Microalgas/metabolismo , Samário/metabolismo , Poluentes Químicos da Água/metabolismo , Transporte Biológico , Chlamydomonas reinhardtii , Células Clonais/química , Metais/metabolismo , Metais Terras Raras/análiseRESUMO
Anthropogenic extreme environments are emphasized as interesting sites for the study of evolutionary pathways, biodiversity, and extremophile bioprospection. Organisms that grow under these conditions are usually regarded as extremophiles; however, the extreme novelty of these environments may have favor adaptive radiations of facultative extremophiles. At the Iberian Peninsula, uranium mining operations have rendered highly polluted extreme environments in multiple locations. In this study, we examined the phytoplankton diversity, community structure, and possible determining factors in separate uranium mining-impacted waters. Some of these human-induced extreme environments may be able to sustain indigenous facultative extremophile phytoplankton species, as well as alleged obligate extremophiles. Therefore, we investigated the adaptation capacity of three laboratory strains, two Chlamydomonas reinhardtii and a Dictyosphaerium chlorelloides, to uranium-polluted waters. The biodiversity among the sampled waters was very low, and despite presenting unique taxonomic records, ecological patterns can be identified. The microalgae adaptation experiments indicated a gradient of ecological novelty and different phenomena of adaptation, from acclimation in some waters to non-adaptation in the harshest anthropogenic environment. Certainly, phytoplankton extremophiles might have been often overlooked, and the ability to flourish in extreme environments might be a functional feature in some neutrophilic species. Evolutionary biology and microbial biodiversity can benefit the study of recently evolved systems such as uranium-polluted waters. Moreover, anthropogenic extremophiles can be harnessed for industrial applications.
Assuntos
Clorófitas/fisiologia , Extremófilos/fisiologia , Fitoplâncton/fisiologia , Urânio/análise , Poluentes Radioativos da Água/análise , Biodiversidade , Chlamydomonas reinhardtii/fisiologia , Chlamydomonas reinhardtii/efeitos da radiação , Clorófitas/efeitos da radiação , Extremófilos/efeitos da radiação , Mineração , Fitoplâncton/efeitos da radiação , Portugal , EspanhaRESUMO
Copper is one of the most frequently used algaecides to control blooms of toxic cyanobacteria in water supply reservoirs. Among the negative impacts derived from the use of this substance is the increasing resistance of cyanobacteria to copper toxicity, as well as changes in the community structure of native phytoplankton. Here, we used the ratchet protocol to investigate the differential evolution and maximum adaptation capacity of selected freshwater phytoplankton species to the exposure of increasing doses of copper. Initially, a dose of 2.5 µM CuSO4·5H2O was able to completely inhibit growth in three strains of the toxic cyanobacterium Microcystis aeruginosa, whereas growth of the chlorophyceans Dictyosphaerium chlorelloides and Desmodesmus intermedius (represented by two different strains) was completely abolished at 12 µM. A significant increase in resistance was achieved in all derived populations during the ratchet experiment. All the chlorophyceans were able to adapt to up to 270 µM of copper sulfate, but 10 µM was the highest concentration that M. aeruginosa strains were able to cope with, although one of the replicates adapted to 30 µM. The recurrent use and increasing doses of copper in water reservoirs could lead to the selection of copper-resistant mutants of both chlorophyceans and cyanobacteria. However, under high concentrations of copper, the composition of phytoplankton community could undergo a drastic change with cyanobacteria being replaced by copper-resistant chlorophyceans. This result stems from a distinct evolutionary potential of these species to adapt to this substance.
Assuntos
Adaptação Fisiológica , Cobre/metabolismo , Fitoplâncton/metabolismo , Poluentes Químicos da Água/metabolismo , Clorófitas , Cianobactérias/genética , Água Doce , Microcystis/genética , Fitoplâncton/genéticaRESUMO
Los Baños de Vilo (S Spain) is a natural spa characterized by extreme sulphureous waters; however, populations of chlorophyceans inhabit in the spa. The adaptation mechanisms allowing resistance by photosynthetic microorganisms to the extreme sulphureous waters were studied by using a modified Luria-Delbrück fluctuation analysis. For this purpose, the adaptation of the chlorophycean Dictyosphaerium chlorelloides and the cyanobacterium Microcystis aeruginosa (both isolated from non-sulphureous water) were analysed in order to distinguish between physiological adaptation (acclimation) and genetic adaptation by the selection of rare spontaneous mutations. Acclimation to the extreme water was achieved by D. chlorelloides; however, M. aeruginosa cells proliferated as a consequence of selection of favoured mutants (i.e. genetic adaptation). The resistant cells of M. aeruginosa appeared with a frequency of 7.1 × 10(-7) per cell per generation, and the frequency of the resistant allele, under non-selective conditions, was estimated to be 1.1 × 10(-6) per cells as a consequence of the balance mutation-selection. It could be hypothesized that the populations of eukaryotic algae living in the Los Baños de Vilo could be the descendants of chlorophyceans that arrived fortuitously at the spa in the past. On the other hand, cyanobacteria could quickly adapt by the selection of favoured mutants. The single mutation that allows resistance to sulphureous water from Baños de Vilo in M. aeruginosa represents a phenotypic burden impairing growth rate and photosynthetic performance. The resistant-variant cells of M. aeruginosa showed a lower acclimated growth rate and a decreased maximum quantum yield and photosynthetic efficiency, in comparison to the wild-type cells.
Assuntos
Clorófitas/fisiologia , Microcystis/metabolismo , Fotossíntese , Sulfetos/metabolismo , Adaptação Fisiológica , Clorófitas/genética , Microcystis/genética , Mutação , Seleção Genética , Espanha , Sulfetos/análise , Água/análiseRESUMO
Future energy supply needs to overcome two challenges: environmental impact and dependence on geopolitically unstable countries. A very promising alternative is based on lithium, an element for batteries, and whose isotope 6Li will be essential in nuclear fusion. The objective of this research has been to determine if it is possible to achieve isotopic fractionation of lithium through a process mediated by microalgae. For this purpose, Chlamydomonas reinhardtii was selected and grown in presence of 5 mg/L of lithium. Results revealed that this specie survives at the selected lithium concentration, discriminates isotopes and preferentially capture 6Li (6δ = 10.029 ± 3.307) through a process independent of the cellular growth. Concomitate recovered up 0.206 mg/L of lithium along a process of 21 days. The result of this study lets to affirm that Chlamydomonas reinhardtii might be used to obtain lithium enriched in the lighter isotope.
RESUMO
The temporal variability of hydrological variables in the Guadalquivir estuary was examined during three years through a real-time remote monitoring network (RTRM). The network was developed with the aim of studying the influence of hydrodynamical and hydrological features within the estuary on the functioning of the pelagic ecosystem. Completing this data-gathering network, monthly cruises were performed in order to measure biogeochemical variables that are indicative of the trophic status of the aquatic environment. The results showed that several sources of physical forcing, such as wind, tide-associated currents and river discharge were responsible for the spatio-temporal patterns of dissolved oxygen, salinity and turbidity in the estuary. The analysis was conducted under tidal and flood regime, which allowed us to identify river discharge as the main forcing agent of the hydrology inside the estuary. In particular, episodes of elevated turbidity detected by the network, together with episodes of low salinity and dissolved oxygen were closely related to the increase in water supply from a dam located upstream. The network installed provided accurate data that can be rapidly used for research or educational applications and by policy-makers or agencies in charge of the management of the coastal area.
Assuntos
Redes de Comunicação de Computadores/instrumentação , Monitoramento Ambiental/instrumentação , Tecnologia de Sensoriamento Remoto/instrumentação , Rios/química , Transdutores , Poluentes Químicos da Água/análise , Água/química , Sistemas Computacionais , Desenho de Equipamento , Análise de Falha de Equipamento , Espanha , Água/análiseRESUMO
Electronic scraps (e-scraps) represent an attractive raw material to mine demanded metals, as well as rare earth elements (REEs). A sequential microbial-mediated process developed in two steps was examined to recover multiple elements. First, we made use of an acidophilic bacteria consortium, mainly composed of Acidiphilium multivorum and Leptospidillum ferriphilum, isolated from acid mine drainages. The consortium was inoculated in a dissolution of e-scraps powder and cultured for 15 days. Forty-five elements were analyzed in the liquid phase over time, including silver, gold, and 15 REEs. The bioleaching efficiencies of the consortium were >99% for Cu, Co, Al, and Zn, 53% for Cd, and around 10% for Cr and Li on Day 7. The second step consisted of a microalgae-mediated uptake from e-scraps leachate. The strains used were two acidophilic extremotolerant microalgae, Euglena sp. (EugVP) and Chlamydomonas sp. (ChlSG) strains, isolated from the same extreme environment. Up to 7.3, 4.1, 1.3, and 0.7 µg by wet biomass (WB) of Zn, Al, Cu, and Mn, respectively, were uptaken by ChlSG biomass in 12 days, presenting higher efficiency than EugVP. Concerning REEs, ChlSG biouptake 14.9, 20.3, 13.7, 8.3 ng of Gd, Pr, Ce, La per WB. Meanwhile, EugVP captured 1.1, 1.5, 1.4, and 7.5, respectively. This paper shows the potential of a microbial sequential process to revalorize e-scraps and recover metals and REEs, harnessing extremotolerant microorganisms.
Assuntos
Acidiphilium/metabolismo , Bactérias/metabolismo , Metais/metabolismo , Mineração/métodos , Reciclagem/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Resíduos Industriais/análise , Microscopia Eletrônica de VarreduraRESUMO
Although the consequences of global warming in aquatic ecosystems are only beginning to be revealed, a key to forecasting the impact on aquatic communities is an understanding of individual species' vulnerability to increased temperature. Despite their microscopic size, phytoplankton support about half of the global primary production, drive essential biogeochemical cycles and represent the basis of the aquatic food web. At present, it is known that phytoplankton are important targets and, consequently, harbingers of climate change in aquatic systems. Therefore, investigating the capacity of phytoplankton to adapt to the predicted warming has become a relevant issue. However, considering the polyphyletic complexity of the phytoplankton community, different responses to increased temperature are expected. We experimentally tested the effects of warming on 12 species of phytoplankton isolated from a variety of environments by using a mechanistic approach able to assess evolutionary adaptation (the so-called ratchet technique). We found different degrees of tolerance to temperature rises and an interspecific capacity for genetic adaptation. The thermal resistance level reached by each species is discussed in relation to their respective original habitats. Our study additionally provides evidence on the most resistant phytoplankton groups in a future warming scenario.
Assuntos
Evolução Biológica , Mudança Climática , Fitoplâncton/genética , Adaptação Fisiológica , Ecossistema , Efeito Estufa , Temperatura Alta , Fitoplâncton/crescimento & desenvolvimento , Fitoplâncton/fisiologia , Especificidade da EspécieRESUMO
Toxic blooms of the cyanobacterium Microcystis aeruginosa affect humans and animals in inland water systems worldwide, and it has been hypothesized that the development of these blooms will increase under the future scenario of global change, considering eutrophication and temperature increase as two important consequences. The importance of genetic adaptation, chance and history on evolution of growth rate, and toxin production of M. aeruginosa was studied under these new conditions. The experiment followed the idea of "replaying life's tape" by means of the simultaneous propagation of 15 independent isolates of three M. aeruginosa strains, which were grown under doubled nutrient concentration and temperature during c. 87 generations. Adaptation by new mutations that resulted in the enhancement of growth rate arose during propagation of derived cultures under the new environmental conditions was the main component of evolution; however, chance also contributed in a lesser extension to evolution of growth rate. Mutations were selected, displacing the wild-type ancestral genotypes. In contrast, the effect of selection on mutations affecting microcystin production was neutral. Chance and history were the pacemakers in evolution of toxin production. Although this study might be considered an oversimplification of the reality, it suggest that a future scenario of global change might lead to an increase in M. aeruginosa bloom frequency, but no predictions about the frequency of toxicity can be made.
Assuntos
Evolução Biológica , Eutrofização , Microcistinas/biossíntese , Microcystis/crescimento & desenvolvimento , Temperatura , Aclimatação , Proliferação Nociva de Algas , Microcystis/química , Microcystis/genética , Mutação , Nitratos/metabolismo , Seleção GenéticaRESUMO
Lithium isotopes are essential for nuclear energy, but new enrichment methods are required. In this study, we considered biotechnology as a possibility. We assessed the Li fractionation capabilities of three Chlorophyte strains: Chlamydomonas reinhardtii, Tetraselmis mediterranea, and a freshwater Chlorophyte, Desmodesmus sp. These species were cultured in Li containing media and were analysed just after inoculation and after 3, 12, and 27 days. Li mass was determined using a Inductively Coupled Plasma Mass Spectrometer, and the isotope compositions were measured on a Thermo Element XR Inductively Coupled Plasma Mass Spectrometer. The maximum Li capture was observed at day 27 with C. reinhardtii (31.66 µg/g). Desmodesmus sp. reached the greatest Li fractionation, (δ6 = 85.4). All strains fractionated preferentially towards 6Li. More studies are required to find fitter species and to establish the optimal conditions for Li capture and fractionation. Nevertheless, this is the first step for a microalgal nuclear biotechnology.
RESUMO
Metals are often spilled by industries into inland water environments, with adverse consequences. Numerous papers have reported that heavy metals produce massive destruction of algae. Nevertheless, algal populations seem to become tolerant when they have had previous exposures to heavy metals. Because the mechanisms allowing heavy metal tolerance of algae are not yet known, the present study analyzed the effect of hexavalent chromium on growth and photosynthetic performance of Dictyosphaerium chlorelloides, stressing on the adaptation mechanisms to chromium contamination. Growth and photosynthetic performance of algal cells were inhibited by Cr(VI) at 10 mg/L, and the 72-h median inhibition concentration was established as 1.64 and 1.54 mg/L, respectively. However, after further incubation for a three month period in an environment with 25 mg/L of chromium, some rare, chromium-resistant cells occasionally were found. A Luria-Delbrück fluctuation analysis was performed to distinguish between resistant algae arising from rare, spontaneous mutations and resistant algae arising from physiological adaptation and other adaptive mechanisms. Resistant cells arose only by spontaneous mutations before the addition of chromium, with a rate of 1.77 x 10(-6) mutants per cell division. From a practical point of view, the use of both chromium-sensitive and chromium-resistant genotypes could make possible a specific algal biosensor for chromium.
Assuntos
Cromo/toxicidade , Eucariotos/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Adaptação Fisiológica , Eucariotos/fisiologia , Fotossíntese/efeitos dos fármacosRESUMO
Resulting from the nuclear fuel cycle, large amounts of depleted uranium (DU) tails are piling up, waiting for possible use or final disposal. To date, the recovery of the residual 235U isotope contained in DU has been conducted only marginally by physical processes. Relative isotope abundances are often mediated by biological processes, and the biologically driven U isotopic fractionation has been previously identified in reducing bacteria. Our results indicate that the cells of two microalgal strains (freshwater Chlamydomonas sp. (ChlGS) and marine Tetraselmis mediterranea (TmmRU)) took up DU from the exposure solutions, inducing U isotopic fractionation with a preference for the fissile 235U isotope over 238U. The n(235U)/n(238U) isotopic fractionation magnitudes (δ235) were 23.6 ± 12.5 and 370.4 ± 103.9, respectively. These results open up new perspectives on the re-enrichment of DU tailings, offering a potential biological alternative to obtain reprocessed natural-equivalent uranium. Additionally, the findings present implications for identifying biological signatures in the geologic records.
Assuntos
Fracionamento Químico/métodos , Chlamydomonas/metabolismo , Clorófitas/metabolismo , Microalgas/metabolismo , Resíduos Radioativos/análise , Urânio/química , Biodegradação Ambiental , Chlamydomonas/classificação , Clorófitas/classificação , Centrais NuclearesRESUMO
Geothermal waters often support remarkable communities of microalgae and cyanobacteria apparently living at the extreme limits of their tolerance. Little is known about the mechanisms allowing adaptation of mesophilic phytoplankters to such extreme conditions, but recent studies are challenging many preconceived notions about this. The aim of this study was to analyse mechanisms allowing adaptation of mesophilic microalgae and cyanobacteria to stressful geothermal waters. To distinguish between the pre-selective or post-selective origin of adaptation processes allowing the proliferation of mesophilic phytoplankters in geothermal waters, several Luria-Delbrück fluctuation analysis were performed with the microalga Dictyosphaerium chlorelloides and the cyanobacterium Microcystis aeruginosa, both isolated from nonextreme waters. Geothermal waters from seven places in Italy and five icebound places at Los Andes in Argentina were used as selective agents. Physiological adaptation was achieved in the least toxic waters. In contrast, rapid genetic adaptation was observed in waters ostensibly lethal for the experimental organisms. This adaptation was achieved as consequence of single mutations at one locus. It was hypothesized that a similar mechanism of rapid genetic adaptation could explain the survival of photosynthetic life during the Neoproterozoic 'snowball Earth,' where geothermal refuges such as those studied could have been 'Noah's Arks' for microalgae and cyanobacteria.
Assuntos
Adaptação Biológica , Clorófitas/genética , Evolução Molecular , Fontes Termais/microbiologia , Microcystis/genética , Fitoplâncton/genética , Clorófitas/fisiologia , Ecossistema , Água Doce/microbiologia , Camada de Gelo , Microcystis/fisiologia , Mutação , Fitoplâncton/fisiologia , Seleção Genética , Estresse FisiológicoRESUMO
Several species of microalgae, closely related to mesophilic lineages, inhabit the extreme environment (pH 2.5, high levels of metals) of the Spain's Aguas Agrias Stream water (AASW). Consequently, AASW constitutes an interesting natural laboratory for analysis of adaptation by microalgae to extremely stressful conditions. To distinguish between the pre-selective or post-selective origin of adaptation processes allowing the existence of microalgae in AASW, a Luria-Delbrück fluctuation analysis was performed with the chlorophycean Dictyosphaerium chlorelloides isolated from non-acidic waters. In the analysis, AASW was used as selective factor. Preselective, resistant D. chlorelloides cells appeared with a frequency of 1.1 x 10(-6) per cell per generation. AASW-resistant mutants, with a diminished Malthusian fitness, are maintained in non-extreme waters as the result of a balance between new AASW-resistant cells arising by mutation and AASW-resistant mutants eliminated by natural selection (equilibrium at c. 12 AASW-resistants per 10(7) wild-type cells). We propose that the microalgae inhabiting this stressful environment could be the descendents of chance mutants that arrived in the past or are even arriving at the present.
Assuntos
Adaptação Fisiológica/fisiologia , Metais/toxicidade , Mineração , Mutação/genética , Mutação/fisiologia , Fitoplâncton/fisiologia , Poluentes Químicos da Água/toxicidade , Poluição da Água/efeitos adversos , Ácidos/toxicidade , Adaptação Fisiológica/genética , Água Doce , Fitoplâncton/genética , Espanha , Testes de ToxicidadeRESUMO
This paper describes the first isolation of Serratia fonticola in a Nile Crocodile (Crocodylus niloticus). This organism was initially isolated from skin lesions and blood and subsequently from a variety of organs during necropsy. S. fonticola was confirmed as the pathogen causing the infection.
Assuntos
Jacarés e Crocodilos/microbiologia , Bacteriemia/veterinária , Infecções por Serratia/veterinária , Serratia/isolamento & purificação , Dermatopatias Bacterianas/veterinária , Animais , Bacteriemia/imunologia , Bacteriemia/microbiologia , DNA Viral/química , DNA Viral/genética , Evolução Fatal , Feminino , Reação em Cadeia da Polimerase/veterinária , Análise de Sequência de DNA , Serratia/genética , Infecções por Serratia/microbiologia , Dermatopatias Bacterianas/microbiologiaRESUMO
The extraction and processing of uranium (U) have polluted large areas worldwide, rendering anthropogenic extreme environments inhospitable to most species. Noticeably, these sites are of great interest for taxonomical and applied bioprospection of extremotolerant species successfully adapted to U tailings contamination. As an example, in this work we have studied a microalgae species that inhabits extreme U tailings ponds at the Saelices mining site (Salamanca, Spain), characterized as acidic (pH between 3 and 4), radioactive (around 4 µSv h-1) and contaminated with metals, mainly U (from 25 to 48 mg L-1) and zinc (from 17 to 87 mg L-1). After isolation of the extremotolerant ChlSP strain, morphological characterization and internal transcribed spacer (ITS)-5.8S gene sequences placed it in the Chlamydomonadaceae, but BLAST analyses identity values, against the nucleotide datasets at the NCBI database, were very low (<92%). We subjected the ChlSP strain to an artificial selection protocol to increase the U uptake and investigated its response to selection. The ancestral strain ChlSP showed a U-uptake capacity of ≈4.30 mg U g-1 of dry biomass (DB). However, the artificially selected strain ChlSG was able to take up a total of ≈6.34 mg U g-1 DB, close to the theoretical maximum response (≈7.9 mg U g-1 DB). The selected ChlSG strain showed two possible U-uptake mechanisms: the greatest proportion by biosorption onto cell walls (ca. 90%), and only a very small quantity, ~0.46 mg g-1 DB, irreversibly bound by bioaccumulation. Additionally, the kinetics of the U-uptake process were characterized during a microalgae growth curve; ChlSG cells removed close to 4 mg L-1 of U in 24 days. These findings open up promising prospects for sustainable management of U tailings waters based on newly evolved extremotolerants and outline the potential of artificial selection in the improvement of desired features in microalgae by experimental adaptation and selection.
RESUMO
Although populations of cyanobacteria are usually considered to be clonal, their capacity to survive environmental changes suggests intrapopulation genetic variation. We therefore estimated the genetic variability on the basis of two processes important for any photoautotroph - photochemical and nonphotochemical quenching - as well as photosynthetic pigment concentrations. For this purpose, two parameters related to photochemical and nonphotochemical quenching were measured using specific experimental and statistical procedures, in 25 strains of the cyanobacterium Microcystis aeruginosa, along with their contents of chlorophyll a, total carotenoids and phycocyanin. The experimental procedure allowed discrimination between genetic and nongenetic (or residual) variability among strains. The high genetic variability found in photosynthetic pigments and both photosynthetic parameters denotes large differences even among strains isolated from the same community. The high genetic diversity within a population could be important for the evolutionary success of cyanobacteria.
Assuntos
Variação Genética , Microcystis/genética , Fotossíntese , Carotenoides/genética , Carotenoides/metabolismo , Clorofila/genética , Clorofila/metabolismo , Microcystis/metabolismo , Ficocianina/genética , Ficocianina/metabolismoRESUMO
A new genetic approach was developed for increasing specificity of microalgal biosensors. This method is based on the use of two different genotypes jointly to detect a given pollutant: (i) a sensitive genotype to obtain sensitivity; and (ii) a resistant mutant to obtain specificity. The method was tested by the development of a microalgal biosensor for the detection of the explosive 2,4,6-trinitrotoluene (TNT) using a wild-type strain (DcG1wt) of Dictyosphaerium chlorelloides (Chlorophyceae) as the sensitive organism, and a TNT-resistant mutant, obtained from DcG1wt strain by a modified Luria-Delbrück fluctuation analysis. The inhibition of chlorophyll a fluorescence of PSII by TNT was used as the biological signal. Significant differences in maximal fluorescence of light-adapted algae (F'(m)) between wild-type DcG1wt cells and TNT-resistant mutants, were observed in all the TNT concentrations tested (from 0.5 to 31.3 mg l(-1)) after only 3 min of exposure. Resistant mutants always exhibited significant higher F'(m) values in the presence of TNT than wild-type cells. These results suggest that the use of two different genotypes (sensitive and resistant to a given pollutant) jointly is a useful method to improve microalgal biosensors specificity.
Assuntos
Técnicas Biossensoriais/métodos , Eucariotos , Trinitrotolueno/análise , Eucariotos/genética , MutaçãoRESUMO
Freshwater toxic cyanobacteria are an increasing problem to human and animal health. Control of cyanobacteria in water supply reservoirs involves the use of algaecides, such as copper sulphate, usually in a repetitive way. Repercussions of recurrent algaecide treatments on cyanobacteria population dynamics remain still unknown. We studied the adaptation of cyanobacteria to lethal doses of copper sulphate by using Microcystis aeruginosa as an experimental model. A fluctuation analysis demonstrated that copper-resistant cells arise by spontaneous mutations that occur randomly prior to exposition to copper sulphate. The rate of spontaneous mutation from copper sensitivity to resistance was 1.76 x 10(-6) mutants per cell division. Resistant mutants exhibited a diminished fitness in the absence of copper sulphate, but only these variants were able to grow at Cu(2+) concentrations higher than 5.8 microM. In addition, copper-resistant cells were significantly smaller than wild-type ones. Warnings on the long-term consequences of repetitive algaecide treatments in water supplies are suggested.