Browsing by Subject "cyanobacteria"

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  • Jäppinen, Sanni (Helsingfors universitet, 2013)
    Filamentous cyanobacteria taxa Nostocales and Stigonematales cells can differentiate into heterocysts nitrogen fixing cells when nitrogen is limiting the growth and into resting cells akinetes when nutrients decrease or the growth conditions become unfavorable for growth. Akinetes overwinter in the water sediments during the unfavorable growth time. When the growing condition improves akinetes germinate and can start a new cyanobacterial bloom. Akinete differentiation remains unclear. It is known from the literature that only a few akinete specific genes exist. First described akinete specific gene was avak. The morphological changes of akinete differentiation are known but the changes at molecular genetics level in regulation and differentiation remains unclear. The aim of this study was to design a method for akinete differentiation-related genes, avak, hepA and hap, for an Anabaena 1TU33s10 strain and to monitor the gene expression changes in a seven-week growth experiment. Primers for the differentiation related genes were designed based on the known whole genome sequence of the Anabaena 1TU3310 strain. In this study it was managed to design a quantitative reverse transcriptase polymerase chain reaction, qRT-PCR method, based on the genes involved in the akinete differentiation process. It was observed that gene expression changed when akinetes began to differentiate into the filaments. In the growth experiment II avaK-gene expression was increased 2-fold between the 14. and 30. days, and hap-gene showed 1.5 fold growth between 14. and 30. days. The number of akinetes was also increasing at the same time. In the growth experiment I heap-gene showed 1-8 fold growth between the days 21. and 27. –30. days when the number of heterocysts were also increasing. The number of akinetes was relatively low compared to number of vegetative cells which also explains the small expression fold-differences in the cultures during the experiment time when compared to expression fold-differences described in the literature. Designed method can thus also detect minor changes in gene expression. The designed and built qRT-PCR method can be used in the future for monitoring gene expression changes also for new akinete specific genes, and the method can be further optimized for screening natural water samples.
  • Kivilompolo, Sanna-Kaisa (Helsingin yliopisto, 2021)
    Aims of this study. Previous studies have shown cyanobacterial dominance and harmful cyanobacterial blooms to increase due to recent climate warming. The increase of aggressively blooming species and toxin-producing strains of cyanobacteria has been predicted to further increase in the future. However, information on the response of cyanobacteria communities to environmental forcing in the Arctic region – which is experiencing warming at over twice the rate compared to the global average – has been insufficient. Thus, it is crucial to study how algal and cyanobacterial communities have developed after industrialization to better understand and predict future trends of subarctic algal communities as well as changes within cyanobacteria communities experiencing environmental forcing. This study aims to provide information on the effect of recent climate warming and lake browning on algal communities in subarctic lakes, with a special focus on cyanobacteria and cyanotoxins. Materials and methods. Modern and historical primary producer group abundances of 23 subarctic lakes located on an ideal temperature and vegetation gradient were studied using sedimentary algal pigments as a proxy. The top-bottom method was used to study both changes within algal communities during the last ca. 150 years and the broader trends in algal communities of subarctic lakes. Pigment data was analyzed together with environmental data using ordination analyses (principal component analysis (PCA) and redundancy analysis (RDA)) as well as other statistical analyses in order to determine possible trends of change and to reveal the environmental variables that have the strongest impact on cyanobacterial abundance. Results and conclusions. Algal communities have changed during the last ca. 150 years and show a general trend of increased primary production as well as lake browning in the spruce, pine and birch (SPB) vegetation zone. Siliceous algae generally dominate modern algal communities, and relative abundances of cyanobacteria have declined throughout the vegetation gradient. Within the Barren (Ba)- and mountain birch woodland (MBW) vegetation zones, cyanobacteria communities show a marked decline in the abundance of assumed benthic species based on pigment data, and low abundances of planktic picocyanobacteria. However, due to climate warming and lake browning, abundances of cyanobacteria have increased in several sites within the SPB vegetation zone and are suspected to indicate an increase of harmful planktic species. The most significant environmental variables controlling the abundance of cyanobacteria were total phosphorus, temperature and the amount of organic matter. The results highlight the urgent need to mitigate climate warming in order to preserve the unique biota and characteristics of Arctic and subarctic lake ecosystems, and to prevent the possible harmful increase of cyanotoxins in these sensitive ecosystems.
  • Teikari, Jonna (Helsingfors universitet, 2013)
    Cyanobacteria are phototrophic organisms. They usually occur in water but many species also live in terrestrial habitats, e.g. in symbiotic relationships with fungus. Inorganic phosphorus (Pi) is usually considered to be a limiting factor for the growth of cyanobacteria living in water, since cyanobacteria can use only Pi as a direct source of phosphorus. It has been shown that cyanobacteria have pho-regulon similar to that of Escherichia coli. Pho-regulon can transport and assimilate inorganic phosphate. Cyanobacteria usually produce a wide range of bioactive compounds, which can e.g. be toxic or prevent growth of other bacteria, fungi or yeast. Many of these compounds are produced by non-ribosomal peptide synthetases (NRPS). The aim of this study was to investigate changes in Anabaena sp. 90 proteome, and differences in amounts of bioactive compounds produced by the strain, while growing it in media with high (5,5 mg/l) or low (0,05 mg/ml) Pi concentration. Before moving the culture into two different Pi media, phosphorus storages of the culture were emptied by growing the strain in the media without Pi. In this study, 2D differential gel electrophoresis (2D-DIGE) coupled with LC/MS was used to study proteomes of the organism. Bioactive compounds were also analyzed by LC/MS. Anabaena sp. 90 was chosen because of its fully sequenced and annotated genome. The strain has been found to produce microcystins, anabaenopeptins and anabaenopeptilides. Eleven protein spots with significantly increased or decreased protein quantities were identified in the low Pi media. Ten of them were identified as proteins, which participate in bacterial stringent response. Stringent response is activated when culture is achieving the stationary phase. These stringent response proteins participate in the amino-acid metabolism and translation. One of the proteins was shown to be a ribosomal protein. In addition, the identified proteins included ribulose-bisphosphate carboxylase oxygenase (RuBisCO), which had a significantly lower concentration in the cells in low-phosphorous media. There were no significant differences in amounts of bioactive compounds when growing the culture in low and high Pi media. More replicates could be used, when the study of bioactive compounds is repeated.
  • Kokocinski, Mikolaj; Dziga, Dariusz; Antosiak, Adam; Soininen, Janne (2021)
    Bacterioplankton community composition has become the center of research attention inrecent years. Bacteria associated with toxic cyanobacteria blooms have attracted considerable interest.However, little is known about the environmental factors driving the bacteria community, includingthe impact of invasive cyanobacteria. Therefore, our aim has been to determine the relationships be-tween heterotrophic bacteria and phytoplankton community composition across 24 Polish lakes withdifferent contributions of cyanobacteria including the invasive speciesRaphidiopsis raciborskii.Thisanalysis revealed that cyanobacteria were present in 16 lakes, whileR. raciborskiioccurred in 14 lakes.Our results show that bacteria communities differed between lakes dominated by cyanobacteria andlakes with minor contributions of cyanobacteria but did not differ between lakes withR. raciborskiiand other lakes. Physical factors, including water and Secchi depth, were the major drivers of bacteriaand phytoplankton community composition. However, in lakes dominated by cyanobacteria, bacte-rial community composition was also influenced by biotic factors such as the amount ofR. raciborskii,chlorophyll-a and total phytoplankton biomass. Thus, our study provides novel evidence on theinfluence of environmental factors andR. raciborskiion lake bacteria communities.
  • Camarena‐Gómez, María Teresa; Ruiz‐González, Clara; Piiparinen, Jonna; Lipsewers, Tobias; Sobrino, Cristina; Logares, Ramiro; Spilling, Kristian (American Society of Limnology and Oceanography, 2021)
    Limnology and Oceanography 66: 1, 255-271
    In parts of the Baltic Sea, the phytoplankton spring bloom communities, commonly dominated by diatoms, are shifting toward the co-occurrence of diatoms and dinoflagellates. Although phytoplankton are known to shape the composition and function of associated bacterioplankton communities, the potential bacterial responses to such a decrease of diatoms are unknown. Here we explored the changes in bacterial communities and heterotrophic production during the spring bloom in four consecutive spring blooms across several sub-basins of the Baltic Sea and related them to changes in environmental variables and in phytoplankton community structure. The taxonomic structure of bacterioplankton assemblages was partially explained by salinity and temperature but also linked to the phytoplankton community. Higher carbon biomass of the diatoms Achnanthes taeniata, Skeletonema marinoi, Thalassiosira levanderi, and Chaetoceros spp. was associated with more diverse bacterial communities dominated by copiotrophic bacteria (Flavobacteriia, Gammaproteobacteria, and Betaproteobacteria) and higher bacterial production. During dinoflagellate dominance, bacterial production was low and bacterial communities were dominated by Alphaproteobacteria, mainly SAR11. Our results suggest that increases in dinoflagellate abundance during the spring bloom will largely affect the structuring and functioning of the associated bacterial communities. This could decrease pelagic remineralization of organic matter and possibly affect the bacterial grazers communities.
  • Teikari, Jonna E.; Hou, Shengwei; Wahlsten, Matti; Hess, Wolfgang R.; Sivonen, Kaarina (2018)
    Salinity is an important abiotic factor controlling the distribution and abundance of Nodularia spumigena, the dominating diazotrophic and toxic phototroph, in the brackish water cyanobacterial blooms of the Baltic Sea. To expand the available genomic information for brackish water cyanobacteria, we sequenced the isolate Nodularia spurn/germ UHCC 0039 using an Illumina-SMRT hybrid sequencing approach, revealing a chromosome of 5,294,286 base pairs (bp) and a single plasmid of 92,326 bp. Comparative genomics in Nostocales showed pronounced genetic similarity among Nodularia spumigena strains evidencing their short evolutionary history. The studied Baltic Sea strains share similar sets of CRISPR-Cas cassettes and a higher number of insertion sequence (IS) elements compared to Nodularia spumigena CENA596 isolated from a shrimp production pond in Brazil. Nodularia spumigena UHCC 0039 proliferated similarly at three tested salinities, whereas the lack of salt inhibited its growth and triggered transcriptome remodeling, including the up-regulation of five sigma factors and the down-regulation of two other sigma factors, one of which is specific for strain UHCC 0039. Down-regulated genes additionally included a large genetic region for the synthesis of two yet unidentified natural products. Our results indicate a remarkable plasticity of the Nodularia salinity acclimation, and thus salinity strongly impacts the intensity and distribution of cyanobacterial blooms in the Baltic Sea.
  • Salmi, Pauliina; Mäki, Anita; Mikkonen, Anu; Pupponen, Veli-Mikko; Vuorio, Kristiina; Tiirola, Marja (Suomen ympäristökeskus, 2021)
    Boreal Environment Research 26: 17-27
    The smaller the phytoplankton, the greater effort is required to distinguish individual cells by optics-based methods. Flow cytometry is widely applied in marine picophytoplankton research, but in freshwater research its role has remained minor. We compared epifluorescence microscopy and flow cytometry in assessing the composition, abundance and cell sizes of autofluorescent picophytoplankton in epilimnia of 46 Finnish lakes. Phycocyaninrich picocyanobacteria were the most dominant. The two methods yielded comparable total picophytoplankton abundances, but the determination of cell sizes, and thus total biomasses, were on average an order of magnitude higher in the microscopy results. However, flow cytometry yielded higher cell sizes when applied on small-celled cultured algae. Our study demonstrated that both epifluorescence microscopy and flow cytometry are useful methods in assessing abundances of phycocyanin-rich and phycoerythrin-rich picocyanobacteria and eukaryotic picophytoplankton in lakes. However, accurate determination of cell size and biomass remain challenges for microscopy and especially for flow cytometry.
  • Humisto, Anu; Herfindal, Lars; Jokela, Jouni; Karkman, Antti; Bjørnstad, Ronja; Choudhury, Romi R.; Sivonen, Kaarina (2015)
    Cyanobacteria are an inspiring source of bioactive secondary metabolites. These bioactive agents are a diverse group of compounds which are varying in their bioactive targets, the mechanisms of action, and chemical structures. Cyanobacteria from various environments, especially marine benthic cyanobacteria, are found to be rich sources for the search for novel bioactive compounds. Several compounds with anticancer activities have been discovered from cyanobacteria and some of these have succeeded to enter the clinical trials. Varying anticancer agents are needed to overcome increasing challenges in cancer treatments. Different search methods are used to reveal anticancer compounds from natural products, but cell based methods are the most common. Cyanobacterial bioactive compounds as agents against acute myeloid leukemia are not well studied. Here we examined our new results combined with previous studies of anti-leukemic compounds from cyanobacteria with emphasis to reveal common features in strains producing such activity. We report that cyanobacteria harbor specific anti-leukemic compounds since several studied strains induced apoptosis against AML cells but were inactive against non-malignant cells like hepatocytes. We noted that particularly benthic strains from the Baltic Sea, such as Anabaena sp., were especially potential AML apoptosis inducers. Taken together, this review and re-analysis of data demonstrates the power of maintaining large culture collections for the search for novel bioactivities, and also how anti-AML activity in cyanobacteria can be revealed by relatively simple and low-cost assays.
  • Liu, Liwei; Herfindal, Lars; Jokela, Jouni; Shishido, Tania Keiko; Wahlsten, Matti; Doskeland, Stein Ove; Sivonen, Kaarina (2014)
  • Wood, Steffaney M.; Kremp, Anke; Savela, Henna; Akter, Sultana; Vartti, Vesa-Pekka; Saarni, Saija; Suikkanen, Sanna (Frontiers in Microbiology, 2021)
    Frontiers in Microbiology 12
    Cyanobacteria of the order Nostocales, including Baltic Sea bloom-forming taxa Nodularia spumigena, Aphanizomenon flosaquae, and Dolichospermum spp., produce resting stages, known as akinetes, under unfavorable conditions. These akinetes can persist in the sediment and germinate if favorable conditions return, simultaneously representing past blooms and possibly contributing to future bloom formation. The present study characterized cyanobacterial akinete survival, germination, and potential cyanotoxin production in brackish water sediment archives from coastal and open Gulf of Finland in order to understand recent bloom expansion, akinete persistence, and cyanobacteria life cycles in the northern Baltic Sea. Results showed that cyanobacterial akinetes can persist in and germinate from Northern Baltic Sea sediment up to >40 and >400 years old, at coastal and open-sea locations, respectively. Akinete abundance and viability decreased with age and depth of vertical sediment layers. The detection of potential microcystin and nodularin production from akinetes was minimal and restricted to the surface sediment layers. Phylogenetic analysis of culturable cyanobacteria from the coastal sediment core indicated that most strains likely belonged to the benthic genus Anabaena. Potentially planktonic species of Dolichospermum could only be revived from the near-surface layers of the sediment, corresponding to an estimated age of 1–3 years. Results of germination experiments supported the notion that akinetes do not play an equally significant role in the life cycles of all bloom-forming cyanobacteria in the Baltic Sea. Overall, there was minimal congruence between akinete abundance, cyanotoxin concentration, and the presence of cyanotoxin biosynthetic genes in either sediment core. Further research is recommended to accurately detect and quantify akinetes and cyanotoxin genes from brackish water sediment samples in order to further describe species-specific benthic archives of cyanobacteria.
  • Suikkanen, Sanna; Uusitalo, Laura; Lehtinen, Sirpa; Lehtiniemi, Maiju; Kauppila, Pirkko; Mäkinen, Katja; Kuosa, Harri (Elsevier, 2021)
    Food Webs 28, e00202
    Blooms of cyanobacteria are recurrent phenomena in coastal estuaries. Their maximum abundance coincides with the productive period of zooplankton and pelagic fish. Experimental studies indicate that diazotrophic, i.e. dinitrogen (N2)-fixing cyanobacterial (taxonomic order Nostocales) blooms affect zooplankton, as well as other phytoplankton. We used multidecadal monitoring data from one archipelago station (1992–2013) and ten open sea stations (1979–2013) in the Baltic Sea to explore the potential bottom-up connections between diazotrophic and non-diazotrophic cyanobacteria and phyto- and zooplankton in natural plankton communities. Random forest regression, combined with linear regression analysis showed that the biomass of cyanobacteria (both diazotrophic and non-diazotrophic) was barely connected to any of the phytoplankton and zooplankton variables examined. Instead, physico-chemical variables (salinity, temperature, total phosphorus), as well as spatial and temporal variability seemed to have more significant connections to both phytoplankton and zooplankton variables. Zooplankton variables were also connected to the biomass of phytoplankton groups other than cyanobacteria (such as chrysophytes, cryptophytes and prymnesiophytes), and phytoplankton variables had connections with the biomass of different zooplankton groups, especially copepods. Overall, negative relationships between cyanobacteria and other plankton taxa were scarcer than expected based on previous experimental studies.
  • Ribeiro Moreira de Assumpção, Christine (Helsingin yliopisto, 2017)
    Biochar is a product from the pyrolysis of plant derived-biomass and it is intended to be applied to soil given its potential of carbon sequestration and soil fertility improvement. Some studies also suggest that increasing application rate of biochar has a positive feedback on biological nitrogen fixation (BNF) and on soil microbial biomass. However, these effects are not well known for boreal forests. The purpose of this study was to evaluate the effects of different biochar application rates: 0 t ha-1, 5 t ha-1 and 10 t ha-1 on BNF, on microbial biomass carbon and nitrogen (MBC and MBN), and on moss biomass. The field experiment was established in Juupajoki, Southern Finland in young Scots pine stands. The stands were amended with biochar one year before the measurements took place. BNF was determined using acetylene reduction assay (ARA), and microbial biomass was estimated using chloroform fumigation-direct extraction (CFDE). The microbial biomass samples were incubated at the temperatures: 10 °C, 15 °C and 20 °C. Biochar amendment raised soil pH, whereas no differences were verified for BNF, MBC, MBN, nor for moss biomass. There was, however, variation in the response of N fixation to incubation temperature, and variation in the response of MBC and MBN to the time of measurement. Observed changes in pH are often likely to justify variations in the rates of BNF and MB, however in this study they were not shown to be of significance. It is possible, however that biochar will have a positive effect on soil vegetation as it is incorporated into the soil in the long-term. Although this study focuses on BNF and MB, the findings may well have a bearing on the use of biochar as a tool for C sequestration, since amendment with biochar was demonstrated as neither beneficial nor harmful to the soil biota.
  • Schaedig, Eric (Helsingin yliopisto, 2020)
    The Baltic Sea is a unique and delicate brackish water ecosystem with high primary productivity driven by oceanic biogeochemical cycles of oxygen, iron, silicon, nitrogen and phosphorus. Elevated anthropogenic nutrient loading into the Baltic ecosystem has resulted in a large-scale increase in destructive cyanobacterial blooms in the open Baltic Sea over the past century. The toxic cyanobacterium Nodularia spumigena is a major component of surface blooms in the open Baltic Sea and continues to bloom even after the depletion of phosphate from the surrounding waters. This phenomenon has been attributed to an ability to scavenge phosphorus from recalcitrant sources. However, the exact phosphorus species that sustain N. spumigena growth in the Baltic Sea remain largely unknown. Here, I employ a comparative genomics approach to determine the evolutionary dynamics of phosphorus scavenging in eight strains of N. spumigena and predict the range of phosphorus sources that may support their growth. Then, I test these predictions by growing six strains of N. spumigena on a number of potentially bioavailable phosphorus sources. Among the phosphorus scavenging genes identified by the genomic analysis, putative pathways for the enzymatic degradation of phytic acid, phosphite, and phosphonates were present and highly conserved in the species. Subsequent growth experiments demonstrated that the organism may grow using phytic acid and phosphite, as well as the phosphonates methylphosphonic acid, ethylphosphonic acid, and nitrilotris(methylenephosphonic acid), as sole phosphorus sources. These results indicate that N. spumigena blooms may be supported by several phosphorus sources previously not known to contribute to eutrophication in the Baltic Sea. While additional growth experiments and further research on the environmental prevalence of these compounds are necessary, the findings presented in this study expand our knowledge of how N. spumigena dominates phytoplankton blooms in a phosphorus-scarce environment and may help to inform future eutrophication mitigation efforts in the Baltic Sea region.
  • Cairns, Johannes (Helsingin yliopisto, 2015)
    The thesis begins with an introduction to the characteristics of experimental organisms, cyanobacteria and cyanophages, and their role in the marine biogeochemical cycles and food-webs. Subsequently, the methodology of experimental evolution and models of host-parasite dynamics are presented. The aim of the experimental part is to test predictions concerning the effects of host-parasite interactions on the marine nitrogen cycle, food-webs, and host properties. Methods include batch culture growth experiments, liquid chromatography–mass spectrometry, an optical density based phage resistance assay, plaque assay, and microscopy. To the author’s knowledge, this is the first controlled study that demonstrates that viral lysis of a diazotrophic cyanobacterium results in the release of cellular nitrogen to the environment in a form that fuels phytoplankton growth. However, evolution with the phage alters the effect. These observations highlight the importance of host-parasite interactions in biogeochemical cycles and food-webs. Further, a novel phage resistant host genotype with short filaments compared to other sensitive and resistant genotypes was detected, with increased growth ability but decreased buoyancy. Reduced buoyancy is proposed as a novel fitness cost of resistance. Phage-mediated evolution resulted in increased diversity in host filament length, growth ability, and buoyancy, supporting the hypothesis that parasites act as drivers of host diversity.
  • Teittinen, Anette; Virta, Leena; Li, Mingjia; Wang, Jianjun (2021)
    Islands provide ideal model systems to examine the factors influencing biodiversity, yet knowledge of microbial biodiversity on islands remains scarce. We collected a dataset from 101 rock pools along a freshwater to brackish water transition on islands of the Baltic Sea and investigated the patterns and drivers of community composition and species richness of diatoms, cyanobacteria and non-cyanobacteria bacteria among islands. We also examined whether environmental heterogeneity increased beta diversity and species richness within islands. Among islands, the patterns in community composition were concordant among the microbial groups, with distinct changes along the freshwater-brackish gradient. The patterns in species richness were context-dependent for each microbial group. In general, richness patterns were most strongly associated with nutrient concentrations or the distances to potential sources of immigrants, whereas no positive relationships between ecosystem size and richness were found. Within islands, environmental heterogeneity was positively correlated with the beta diversity of each microbial group, but not species richness. Our findings provide novel insights into the factors influencing microbial biodiversity. The results suggest that island microbial biodiversity patterns are influenced by species sorting and dispersal-related mechanisms and highlight the importance of environmental heterogeneity for beta diversity.
  • Kraft, Kaisa; Seppälä, Jukka; Hällfors, Heidi; Suikkanen, Sanna; Ylöstalo, Pasi; Anglès, Sílvia; Kielosto, Sami; Kuosa, Harri; Laakso, Lauri; Honkanen, Martti; Lehtinen, Sirpa; Oja, Johanna; Tamminen, Timo (Frontiers Media S.A., 2021)
    Frontiers in Marine Science 8: 594144
    Cyanobacteria are an important part of phytoplankton communities, however, they are also known for forming massive blooms with potentially deleterious effects on recreational use, human and animal health, and ecosystem functioning. Emerging high-frequency imaging flow cytometry applications, such as Imaging FlowCytobot (IFCB), are crucial in furthering our understanding of the factors driving bloom dynamics, since these applications provide community composition information at frequencies impossible to attain using conventional monitoring methods. However, the proof of applicability of automated imaging applications for studying dynamics of filamentous cyanobacteria is still scarce. In this study we present the first results of IFCB applied to a Baltic Sea cyanobacterial bloom community using a continuous flow-through setup. Our main aim was to demonstrate the pros and cons of the IFCB in identifying filamentous cyanobacterial taxa and in estimating their biomass. Selected environmental parameters (water temperature, wind speed and salinity) were included, in order to demonstrate the dynamics of the system the cyanobacteria occur in and the possibilities for analyzing high-frequency phytoplankton observations against changes in the environment. In order to compare the IFCB results with conventional monitoring methods, filamentous cyanobacteria were enumerated from water samples using light microscopical analysis. Two common bloom forming filamentous cyanobacteria in the Baltic Sea, Aphanizomenon flosaquae and Dolichospermum spp. dominated the bloom, followed by an increase in Oscillatoriales abundance. The IFCB results compared well with the results of the light microscopical analysis, especially in the case of Dolichospermum. Aphanizomenon biomass varied slightly between the methods and the Oscillatoriales results deviated the most. Bloom formation was initiated as water temperature increased to over 15°C and terminated as the wind speed increased, dispersing the bloom. Community shifts were closely related to movements of the water mass. We demonstrate how using a high-frequency imaging flow cytometry application can help understand the development of cyanobacteria summer blooms.
  • Coloma, Sebastian Ernesto; Gaedge, Ursula; Sivonen, Anna Kaarina; Hiltunen, Teppo Johannes (2019)
    Parasites, such as bacterial viruses (phages), can have large effects on host populations both at the ecological and evolutionary levels. In the case of cyanobacteria, phages can reduce primary production and infected hosts release intracellular nutrients influencing planktonic food web structure, community dynamics, and biogeochemical cycles. Cyanophages may be of great importance in aquatic food webs during large cyanobacterial blooms unless the host population becomes resistant to phage infection. The consequences on plankton community dynamics of the evolution of phage resistance in bloom forming cyanobacterial populations are still poorly studied. Here, we examined the effect of different frequencies of a phage-resistant genotype within a filamentous nitrogen-fixing Nodularia spumigena population on an experimental plankton community. Three Nodularia populations with different initial frequencies (0%, 5%, and 50%) of phage-resistant genotypes were inoculated in separate treatments with the phage 2AV2, the green alga Chlorella vulgaris, and the rotifer Brachionus plicatilis, which formed the experimental plankton community subjected to either nitrogen-limited or nitrogen-rich conditions. We found that the frequency of the phage-resistant Nodularia genotype determined experimental community dynamics. Cyanobacterial populations with a high frequency (50%) of the phage-resistant genotype dominated the cultures despite the presence of phages, retaining most of the intracellular nitrogen in the plankton community. In contrast, populations with low frequencies (0% and 5%) of the phage-resistant genotype were lysed and reduced to extinction by the phage, transferring the intracellular nitrogen held by Nodularia to Chlorella and rotifers, and allowing Chlorella to dominate the communities and rotifers to survive. This study shows that even though phages represent minuscule biomass, they can have key effects on community composition and eco-evolutionary feedbacks in plankton communities.
  • Geraldes, Vanessa; de Medeiros, Livia Soman; Lima, Stella T.; Alvarenga, Danillo Oliveira; Gacesa, Ranko; Long, Paul F.; Fiore, Marli Fatima; Pinto, Ernani (2020)
    Cyanobacteria have been widely reported to produce a variety of UV-absorbing mycosporine-like amino acids (MAAs). Herein, we reported production of the unusual MAA, mycosporine-glycine-alanine (MGA) in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024 using a newly developed UHPLC-DAD-MS/HRMS (ultra-high performance liquid chromatography-diode array detection-high resolution tandem mass spectrometry) method. MGA had previously been first identified in a red-algae, but S. torques-reginae strain ITEP-024 is the first cyanobacteria to be reported as an MGA producer. Herein, the chemical structure of MGA is fully elucidated from one-dimensional / two-dimensional nuclear magnetic resonance and HRMS data analyses. MAAs are unusually produced constitutively in S. torques-reginae ITEP-024, and this production was further enhanced following UV-irradiance. It has been proposed that MAA biosynthesis proceeds in cyanobacteria from the pentose phosphate pathway intermediate sedoheptulose 7-phosphate. Annotation of a gene cluster encoded in the genome sequence of S. torques-reginae ITEP-024 supports these gene products could catalyse the biosynthesis of MAAs. However, addition of glyphosate to cultures of S. torques-reginae ITEP-024 abolished constitutive and ultra-violet radiation induced production of MGA, shinorine and porphyra-334. This finding supports involvement of the shikimic acid pathway in the biosynthesis of MAAs by this species.
  • Arsin, Sila (Helsingin yliopisto, 2019)
    Mycosporines and mycosporine-like amino acids (MAAs) are small-molecules that provide UV protection in a broad range of organisms. Cyanobacteria produce a diverse set of MAA chemical variants, many of which are glycosylated. Even though the biosynthetic pathway for the production of a common cyanobacterial MAA, shinorine, is known, the biosynthetic origins of the glycosylated variants remains unclear. In this work, bioinformatics analyses were performed to catalogue the genetic diversity encoded in the MAA gene clusters in cyanobacterial genomes and identify a set of enzymes that might be involved in MAA biosynthesis. A total of 211 cyanobacterial genomes were found to contain the MAA gene cluster, with six containing glycosyltransferase genes within the gene cluster. Afterwards, 38 strains from the University of Helsinki Culture Collection were tested for the production of MAAs using QTOF-LC/MS analyses. This resulted in the identification of several novel glycosylated MAA chemical variants from Nostoc sp. UHCC 0302, which contained a 7.4 kb MAA biosynthetic gene cluster consisting of 7 genes, including two for glycosyltransferases and one for dioxygenase. Heterologous expression of this gene cluster in Escherichia coli TOP10 resulted in the production of a glycosylated porphyra-334 variant of 509 m/z by the transformant cells, showing that colanic acid biosynthesis glycosyltransferases can catalyse the addition of hexose to MAAs. These results suggested a biosynthetic route for the production of glycosylated MAAs in cyanobacteria and allowed to propose a putative role for dioxygenases in MAA biosynthesis. Further characterization of additional glycosyltransferases is necessary to improve our understanding of glycosylated MAA biosynthesis and functionality, which could be applied to large scale processes and be used in industrial applications.