Browsing by Author "Poonam Bhargava"

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A comparative study of antioxidative defense system in the copper and temperature acclimated strains of Anabaena doliolum
(2008) Yogesh Mishra; Poonam Bhargava; Riti Thapar; Ashish Kumar Srivastava; Lal Chand Rai
This study provides first hand comparative account of growth and antioxidative defense system of the wild type, Cu2+ and temperature treated wild type and acclimated strains of Anabaena doliolum Bharadwaja against Cu2+ and high temperature. The acclimated strains showed perceptible growth at 250 μM Cu2+ and 47°C temperatures, respectively. In contrast to this the wild type strain on exposure to 50 μM Cu2+ and 47°C temperature depicted almost complete inhibition of growth. However, the peroxide content was significantly higher in the acclimated strains than the wild type. Superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) showed maximum activity at high temperature followed by Cu2+ acclimated and minimum in the wild type strains. The ascorbate (ASC) and glutathione (GSH) contents were increased by 2.3 and 43.3, and 15.5 and 36.5-fold in Cu2+ and 47°C acclimated strains, respectively. However, when the wild type strain was subjected to Cu2+ and temperature all antioxidative enzymes except SOD showed inhibition of their activity. In case of wild type the GSH content was inhibited by 0.39-fold at 50 μM Cu2+ but the ASC content registered increase by 2 and 2.7-fold on subjecting to Cu2+ and temperature, respectively. Thus increased activity of enzymatic antioxidants as well as accumulation of ascorbate and glutathione in both the acclimated strains suggests that enzymatic and non-enzymatic antioxidants help in the acclimation of A. doliolum Bharadwaja against Cu2+ and high temperature. However, inhibition of antioxidative defense system of wild type under Cu2+ and heat stress appears to be the reason for its non survival. In view of the appreciable increase in the level of antioxidants as well as greater inhibition of specific growth rate in temperature than Cu2+ acclimated strains, temperature (47°C) is proposed to be is more deleterious to the organism than copper (250 μM). © 2008 Springer Science+Business Media B.V.
A single gene all3940 (Dps) overexpression in Anabaena sp. PCC 7120 confers multiple abiotic stress tolerance via proteomic alterations
(Springer Verlag, 2016) Om Prakash Narayan; Nidhi Kumari; Poonam Bhargava; Hema Rajaram; Lal Chand Rai
DNA-binding proteins (Dps) induced during starvation play an important role in gene regulation and maintaining homeostasis in bacteria. The nitrogen-fixing cyanobacterium, Anabaena PCC7120, has four genes annotated as coding for Dps; however, the information on their physiological roles is limiting. One of the genes coding for Dps, ‘all3940’ was found to be induced under different abiotic stresses in Anabaena and upon overexpression enhanced the tolerance of Anabaena to a multitude of stresses, which included salinity, heat, heavy metals, pesticide, and nutrient starvation. On the other hand, mutation in the gene resulted in decreased growth of Anabaena. The modulation in the levels of All3940 in Anabaena, achieved either by overexpression of the protein or mutation of the gene, resulted in changes in the proteome, which correlated well with the physiological changes observed. Proteins required for varied physiological activities, such as photosynthesis, carbon-metabolism, oxidative stress alleviation, exhibited change in protein profile upon modulation of All3940 levels in Anabaena. This suggested a direct or an indirect effect of All3940 on the expression of the above stress-responsive proteins, thereby enhancing tolerance in Anabaena PCC7120. Thus, All3940, though categorized as a Dps, is possibly a general stress protein having a global role in regulating tolerance to multitude of stresses in Anabaena. © 2015, Springer-Verlag Berlin Heidelberg.
Copper pretreatment augments ultraviolet B toxicity in the cyanobacterium Anabaena doliolum: A proteomic analysis of cell death
(2008) Poonam Bhargava; Arvind Kumar; Yogesh Mishra; Lal Chand Rai
This study provides first-hand proteomic characterisation of Cu-pretreatment-induced augmentation of ultraviolet B toxicity in the cyanobacterium Anabaena doliolum Bharadwaja. Of the three treatments (i.e. Cu, UV-B and Cu:+:UV-B) tested, the UV-B treatment of Cu-pretreated Anabaena produced a greater inhibition of oxygen evolution, 14C fixation, ATP and NADPH contents than UV-B alone. Proteomic analysis using two-dimensional gel electrophoresis (2DE), MALDI-TOF MS/MS and reverse transcription polymerase chain reaction (RT-PCR) of Cu, UV-B, and Cu:+:UV-B treated Anabaena exhibited significant and reproducible alterations in 12 proteins. Of these, manganese superoxide dismutase (Mn-SOD), iron superoxide dismutase (Fe-SOD) and peroxiredoxin (PER) are antioxidative enzymes; ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo), phosphoribulokinase (PRK), flavodoxin (Flv), plastocyanin (PLC), phosphoglycerate kinase (PGK), phycocyanin (PC) and phycoerythrocyanin α-chain (PC α-chain) are linked with photosynthesis and respiration; and DnaK and nucleoside diphosphate kinase (NDPK) are associated with cellular processes and light signalling, respectively. However, when subjected to a high dose of UV-B, Cu-pretreated Anabaena depicted a severe down-regulation of DnaK, NDPK and Flv, probably because of inevitable oxidative stress. Thus, the augmentation of UV-B toxicity by Cu can be attributed to the down-regulation of DnaK, NDPK and Flv. © CSIRO 2008.
Differential induction of enzymes and antioxidants of the antioxidative defense system in Anabaena doliolum exposed to heat stress
(2005) Yogesh Mishra; Poonam Bhargava; Lal Chand Rai
Anabaena doliolum subjected to 43, 48, 53 and 58°C temperature for 1, 2, 3 and 4 h, showed temperature and time-dependent increase in H 2O2 production and MDA contents. All the measured enzymes of the antioxidative defense system (SOD, CAT, APX and GR) showed increase in their activities at 43°C after 1 h of treatment, but at higher temperature their activity declined. The content of antioxidants (ASC, GSH, and α-TOC) increased significantly with rise in temperature as well as duration of treatment. This study clearly demonstrates that when enzymatic defense system becomes inactive, the antioxidants (GSH, and α-TOC) are induced to protect the cyanobacterium from heat stress. One of the major roles of these antioxidants appears to be the protection of PSII as reflected by an effect on O2 evolution up to 53°C. © 2005 Elsevier Ltd. All rights reserved.
Differential response of antioxidative defense system of Anabaena doliolum under arsenite and arsenate stress
(2009) Ashish Kumar Srivastava; Poonam Bhargava; Riti Thapar; Lal Chand Rai
This study offers first hand information on the arsenite (As(III)) and arsenate (As(V))-induced oxidative tress and changes in antioxidative defense system of Anabaena doliolum. A requirement of 58 mM As(V) as compared to only 11 mM As(III) to cause 50% reduction in growth rate suggests that As(III) is more toxic than As(V) in the test cyanobacterium. In contrast to above, oxidative damage measured in terms of lipid peroxidation, electrolyte leakage and peroxide content were significantly higher after As(V) than As(III) treatment as compared to control. Similarly all the studied enzymatic parameters of antioxidative defense system except glutathione reductase (GR) and non-enzymatic parameters except glutathione reduced (GSH) showed greater induction against As(V) than As(III). Interestingly, higher increase in non-enzymatic counterpart than enzymatic in both the stresses suggests that detoxification is mainly managed by former than the later. This confirms the belief of pronounced stimulation of the antioxidative defense system by As(V) than As(III). In conclusion, the cyanobacterium may survive better in As(V) than As(III) contaminated fields because of its low toxicity and pronounced induction of antioxidative defense system. © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Effect of pretreatment of salt, copper and temperature on ultraviolet-B-induced antioxidants in diazotrophic cyanobacterium Anabaena doliolum
(2006) Ashish Kumar Srivastava; Poonam Bhargava; Yogesh Mishra; Bideh Shukla; Lal Chand Rai
Effect of salt, copper, and temperature pretreatments on the UV-B-induced oxidative damage, measured in terms of peroxide and MDA (lipid peroxidation) contents, was studied in the diazotrophic cyanobacterium Anabaena doliolum. To understand the survival strategy enzymatic (superoxide dismutase, catalase, glutathione reductase, and ascorbate peroxidase) and non-enzymatic (glutathione, ascorbate, α-tocopherol and carotenoid) antioxidants were studied. Among the various pretreatments salt was found to decrease and copper and temperature pretreatments increased the deleterious effects of UV-B. This study is the first to demonstrate that physical stress (high temperature) enhanced the damaging effect of UV-B more profoundly than chemical stresses (salt and copper). © 2006 Wiley-VCH Verlag GmbH & Co. KGaA.
Excess copper induces anoxygenic photosynthesis in Anabaena doliolum: A homology based proteomic assessment of its survival strategy
(2008) Poonam Bhargava; Yogesh Mishra; Ashish Kumar Srivastava; Om Prakash Narayan; Lal Chand Rai
This study is the first to demonstrate operation of anoxygenic photosynthesis in copper acclimated Anabaena doliolum and to offer proteomic comparison with the control cells. The Cu-treated control strain showed a negative correlation in growth and intracellular Cu, partial inhibition of O2-evolution, PS II, PS I, whole chain, chlorophyll absorption, and nitrogenase activity. However, the acclimated strain growing in 250-fold excess Cu exhibited near normal growth, ATP content, PS I activity, carbon fixation, and almost complete inhibition of O2-evolution, PS II and chlorophyll absorption, but increased nitrogenase activity as compared to control. Proteomic decoding of the survival strategy of Cu-treated control and the acclimated strain using two-dimensional gel electrophoresis and MALDI-TOF MS analysis of proteins displaying significant and reproducible changes demonstrated involvement of transketolase, phycoerythrocyanin α-chain, iron superoxide dismutase (Fe-SOD), hypothetical protein alr 0803, manganese superoxide dismutase (Mn-SOD), phosphoribulokinase, and plastocyanin (PLC). Expression pattern of these proteins was attested at the transcriptional level using RT-PCR. Time course analysis of proteins of Cu-treated control strain revealed almost no change in PLC level, and a minor accumulation of transketolase, phycoerythrocyanin α-chain and both isoforms of SOD after 7 and recovery after 10 days. Acclimated strain under excess Cu, however, exhibited significant accumulation of both isoforms of SOD, plastocyanin, phosphoribulokinase and transketolase, which seem to counteract oxidative damage, serve as an alternate electron carrier from cytochrome b6/f complex to photosystem I and meet the NADPH and ATP requirements, respectively, under anoxygenic photosynthesis. In view of the kinetics of the hypothetical protein alr0803 (no change in expression level for 7, maximum after 10 and decline after 15 days) its involvement in metal homeostasis is suggested. © 2007 Springer Science+Business Media B.V.
Impact of different abiotic stresses on growth, photosynthetic electron transport chain, nutrient uptake and enzyme activities of Cu-acclimated Anabaena doliolum
(Elsevier GmbH, 2008) Riti Thapar; Ashish Kumar Srivastava; Poonam Bhargava; Yogesh Mishra; Lal Chand Rai
This study provides a comparative account of the effects of cadmium, temperature, ultraviolet-B and sodium chloride on the growth, photosynthesis, nutrient uptake and enzyme activities of untreated control and copper-acclimated Anabaena doliolum. Reduction in all the studied parameters, except carotenoids, was maximum for sodium chloride followed by ultraviolet-B, temperature and cadmium treatments, the reduction being greater in control than acclimated A. doliolum. Among the various parameters, photosystem II was most sensitive for all the stresses in both control and acclimated A. doliolum. Likewise, O2 evolution was more susceptible to various stressors than 14C uptake. Ammonium uptake and glutamine synthetase (GS, EC 6.3.1.2) were the least affected parameters. As compared to control, acclimated Anabaena exhibited higher ATP content under normal conditions. These results attest our hypotheses that acclimated Anabaena was physiologically more robust than control and that salinity was more injurious to the test organism than other abiotic stresses investigated. © 2007 Elsevier GmbH. All rights reserved.
Molecular approaches towards assessment of cyanobacterial biodiversity
(2009) Nidhi Kumari; Ashish Kumar Srivastava; Poonam Bhargava; Lal Chand Rai
Cyanobacteria, a diverse group of photosynthetic prokaryotes, often play central roles in carbon and nitrogen assimilation which makes their environment productive. Despite their significant agronomic values, biodiversity assessment and conservation of these important microbes have not been given much attention by researchers. This review tries to bring together the important aspects of molecular approaches being used for cyanobacterial biodiversity assessment. Molecular approaches have been divided into 2 classes: PCR independent and PCR based approaches. While the former includes guanine plus cytosine (G+C) content, nucleic acid re-association and hybridization and DNA microarrays, the latter basically uses DNA cloning and sequencing, DGGE (denaturing gradient gel electrophoresis), TGGE (temperature gradient gel electrophoresis), SSCP (single strand conformation polymorphism), RFLP (restriction fragment length polymorphism), ARDRA (amplified ribosomal DNA restriction analysis), T-RFLP (terminal restriction fragment length polymorphism), RISA (ribosomal intergenic spacer analysis), ARISA (automated ribosomal intergenic spacer analysis) and RAPD (random amplified polymorphic DNA). Molecular assessment of cyanobacterial biodiversity frequently uses markers like 16S rDNA, phycocyanin locus, nif gene, rpo gene, ITS region etc. This article examines and compares various conventional and modern methods and their short comings, if any. It attempts to provide a general overview of biodiversity assessment, molecular techniques and markers used for biodiversity assessment and also recommends combinatorial approach with different molecular markers. It is likely to improve the degree of resolution and provide as possible the broadest picture and indepth information about biodiversity documentation. © 2009 Academic Journals.
Molecular characterization and the effect of salinity on cyanobacterial diversity in the rice fields of Eastern Uttar Pradesh, India
(2009) Ashish Kumar Srivastava; Poonam Bhargava; Arvind Kumar; Lal Chand Rai; Brett A Neilan
Background. Salinity is known to affect almost half of the world's irrigated lands, especially rice fields. Furthermore, cyanobacteria, one of the critical inhabitants of rice fields have been characterized at molecular level from many different geographical locations. This study, for the first time, has examined the molecular diversity of cyanobacteria inhabiting Indian rice fields which experience various levels of salinity. Results. Ten physicochemical parameters were analyzed for samples collected from twenty experimental sites. Electrical conductivity data were used to classify the soils and to investigate relationship between soil salinity and cyanobacterial diversity. The cyanobacterial communities were analyzed using semi-nested 16S rRNA gene PCR and denaturing gradient gel electrophoresis. Out of 51 DGGE bands selected for sequencing only 31 which showed difference in sequences were subjected to further analysis. BLAST analysis revealed highest similarity for twenty nine of the sequences with cyanobacteria, and the other two to plant plastids. Clusters obtained based on morphological and molecular attributes of cyanobacteria were correlated to soil salinity. Among six different clades, clades 1, 2, 4 and 6 contained cyanobacteria inhabiting normal or low saline (having EC < 4.0 ds m-1) to (high) saline soils (having EC > 4.0 ds m-1), however, clade 5 represented the cyanobacteria inhabiting only saline soils. Whilst, clade 3 contained cyanobacteria from normal soils. The presence of DGGE band corresponding to Aulosira strains were present in large number of soil indicating its wide distribution over a range of salinities, as were Nostoc, Anabaena, and Hapalosiphon although to a lesser extent in the sites studied. Conclusion. Low salinity favored the presence of heterocystous cyanobacteria, while very high salinity mainly supported the growth of non-heterocystous genera. High nitrogen content in the low salt soils is proposed to be a result of reduced ammonia volatilization compared to the high salt soils. Although many environmental factors could potentially determine the microbial community present in these multidimensional ecosystems, changes in the diversity of cyanobacteria in rice fields was correlated to salinity.
Phytochelatin plays a role in UV-B tolerance in N2-fixing cyanobacterium Anabaena doliolum
(Elsevier GmbH, 2005) Poonam Bhargava; Ashish Kumar Srivastava; Snigdha Urmil; Lal Chand Rai
To study the role of Cd-induced phytochelatins in UV-B tolerance, lipid peroxidation, antioxidative enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase), glutathione and phytochelatin contents were measured in buthionine sulphoximine treated and untreated cells of Anabaena doliolum. Cd-pretreatment of the cyanobacterium reduced the lipid peroxidation as well as the antioxidative enzymes in comparison to UV-B treatment alone, whereas the phytochelatin content demonstrated an increase. In contrast to this, buthionine sulphoximine-induced inhibition of phytochelatin synthase, dramatically decreased the Cd-induced co-tolerance against UV-B, hence demonstrating that phytochelatin not only protects the cyanobacterium from heavy metal but participates in UV-B tolerance as well. © 2005 Elsevier GmbH. All rights reserved.
Preliminary analysis of cuprome of Anabaena doliolum using two-dimensional gel electrophoresis
(2006) Poonam Bhargava; Yogesh Mishra; Ashish Kumar Srivastava; Anjum Ara; Lal Chand Rai
This study provides first-hand information on the initial characterization of copper-induced changes in the global proteome (hereafter called cuprome) of Anabaena doliolum subjected to short- and long-term treatments. PD Quest analysis revealed that out of 215 protein spots in the control, 79 showed alterations (26 up- and 36 down-regulation, and 5 up- and 12 down-regulation respectively, after 24 and 168 h of Cu treatment) in their expression pattern. The short-term (24 h) and long-term (168 h) treatments induced respectively, 158 and 96 proteins. Of the 158 newly induced proteins, 30 were found to sustain the long-term treatment. In view of the appearance of two sets of proteins, there appears a need to carry out short- and long-term proteome analysis for getting a holistic view of the proteome of cyanobacteria subjected to abiotic stress.
Protein fingerprinting may serve as a complementary tool for the phylogenetic classification of heterocystous (Nostoc, Anabaena, Cylindrospermum, Aulosira and Tolypothrix) Cyanobacteria
(2010) Swati Mishra; Poonam Bhargava; Rashmi Rai; Yogesh Mishra; Teresa Zotta; Eugenio Parente; Lal Chand Rai
A combination of morphology, SDS-PAGE, 16S rRNA gene and the fuzzy approach has been used for the first time to attest the phylogenetic affiliation of the cyanobacterial species such as Nostoc, Anabaena, Cylindrospermum, Tolypothrix and Aulosira. The idea was to ascertain as how far SDS-PAGE analysis and morphological taxonomy work coherently and serve as a reliable tool, compatible with 16S rRNA gene based classification of cyanobacteria. Intermingling of Nostoc and Anabaena species in SDS-PAGE and 16S rRNA gene indicates that the two taxa are not clearly separated at the genetic level and may be polyphyletic in origin. In contrast to this, clustering of Tolypothrix and Aulosira species in the same clade attests the taxonomic coherence of these species. However, Cylindrospermum showed greater phylogenetic relatedness with Anabaena strain in all phylogenetic analyses. Despite a few minor incongruities, protein fingerprint depicted greater similarity with 16S rRNA gene as compared to morphological taxonomy. Thus protein fingerprinting developed using fuzzy approach appears to be more compatible with 16S rRNA gene analysis than morphological taxonomy in discriminating cyanobacteria up to the species level. Copyright Internet Scientific Publications, LLC., 1996 to 2010.
Proteomic evaluation of the non-survival of Anabaena doliolum (Cyanophyta) at elevated temperatures
(2009) Yogesh Mishra; Poonam Bhargava; Neha Chaurasia; Lal Chand Rai
This paper presents proteomic evidence for the non-survival of Anabaena doliolum at elevated temperatures (43°C, 48°C, 53°C and 58°C), when photosystem II (PSII), carbon fixation, ATP and NADPH contents were significantly decreased. A total of 215,200,261,229 and 99 spots were clearly visible in two-dimensional gel electrophoresis (2DE) gels of the cyanobacterium from the control and those subjected to 1 h treatment at the above temperatures, respectively. Proteomic analysis using 2DE, MALDI-TOF MS/MS and reverse transcription polymerase chain reaction of Anabaena exposed to the above temperatures displayed significant and reproducible alterations in 13 proteins homologous to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), fructose-1, 6-bisphosphate aldolase (FBA), fructose-1, 6-bisphosphatase (FBPase), keto-acid reductoisomerase, phycocyanin-α-chain, peroxiredoxin, ATP synthase-β-chain, RNA binding protein, nucleoside diphosphate kinase (NDPK), GroES, phycoerythrocyanin-α-chain, AhpC/TSA family and phycobilisome rod-core linker (PBS-RCL) protein. Except Prx, GroES and ATP synthase-β-chain, other metabolic and oxidative stress proteins were down-regulated at temperatures over 48 C. The down-regulation of FBA, GAPDH, and FBPase beyond 48 C possibly disrupted glycolysis, the pentose phosphate pathway and the Calvin cycle, thereby leading to the death of Anabaena above 48°C. Notwithstanding the above, an appreciable down-regulation of NDPK (involved in protein phosphorylation) and AhpC/TSA (specific scavenger of reactive sulphur species), may lead to loss of signalling and accumulation of reactive sulphur species, respectively, which contribute further to temperature stress. Appreciable induction of heat-shock proteins (HSPs), required for maintenance of metabolic proteins during thermal stress, as well as metabolic proteins up to 48°C and their down-regulation at 53°C and 58°C suggested a close interlinking of these proteins. However, down-regulation of HSP60, HSP26 and HSP 16.6 appears to be responsible for cell death. © 2009 British Phycological Society.
Salinity and copper-induced oxidative damage and changes in the antioxidative defence systems of Anabaena doliolum
(2005) Ashish Kumar Srivastava; Poonam Bhargava; Lal Chand Rai
This study provides first-hand information on the salinity and copper-induced oxidative damage and its protection in Anabaena doliolum by the antioxidant defence system. Oxidative damage measured in terms of lipid peroxidation, electrolyte leakage and H2O2 production was induced by different concentrations of NaCl and Cu2+. A greater electrolyte leakage by NaCl than Cu2+ supported the hypothesis of salinity being more injurious than copper. To explore the survival strategies of A. doliolum under NaCl and Cu stress, enzymatic antioxidant activities e.g. superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) and nonenzymatic antioxidant contents such as glutathione reduced (GSH), ascorbate, α-tocopherol, and carotenoid were measured. A general induction in SOD and APX activities as well as ascorbate and α-tocopherol contents was found under NaCl and Cu2+ stress. In contrast to this, an appreciable decline in GR activity, GSH pool and carotenoid content under Cu2+ and an increase under NaCl stress were observed. CAT activity was completely inhibited at high doses of NaCl but stimulated following Cu2+ treatment. The above results suggest the involvement of APX and CAT in the scavenging of H2O2 under Cu2+ stress. In contrast to this, only APX was involved in H 2O2 scavenging under salt stress. Our postulate of Cu 2+-mediated antagonism of salt stress can be explained by a conceivable reversion of Na+-induced disturbance of cellular homeostasis by redox active Cu2+. © Springer 2005.
Weighted morphology: a new approach towards phylogenetic assessment of Nostocales (Cyanobacteria)
(Springer-Verlag Wien, 2015) Swati Mishra; Poonam Bhargava; Siba Prasad Adhikary; Anubhav Pradeep; Lal Chand Rai
The classification of order Nostocales (Cyanobacteria) and inter relationships of morphologically similar taxa is still debatable due to ever changing morphological features. No attempt has been made to improve the morphological taxonomy despite the fact that it is the morphology that represents the totality of genes. To test the validity of morphological taxonomy and fine tune the phylogenetic relationships within the order Nostocales a new weighted morphology approach was applied by using 76 isolates and their 16S rRNA gene sequences. Further, the study was extended with morphological data set of the remaining 232 taxa for which no molecular data are yet available. Trichome aggregation, heterocyst shape, and akinete shape are suggested as important and stable features for identification. At 30 % weight assignment to the selected morphological characters, morphological taxonomy found 36 % compatible with 16S tree. Adding weight to the morphological characters considerably improved the congruence between the morphology and 16S rRNA-based phylogenetic trees of the order Nostocales. When the weighting procedure was extended to all the Nostocalean members irrespective of molecular data availability, it was found that Nostoc sphaericum and Nostoc microscopicum closely assembled in a single clade. Closer arrangement of Aulosira and Nodularia represent the subfamily aulosirae (Bornet and Flahault Ann Sci Nat Bot 7:223–224, 1888) while taxonomic affiliation of Cylindrospermum with Nostoc, Anabaena, and Raphidiopsis representing the subfamily anabaenae (Bornet and Flahault Ann Sci Nat Bot 7:223–224, 1888) was resolved. © 2014, Springer-Verlag Wien.