Igniting taxonomic curiosity: The amazing story of Amazonocrinis with the description of a new genus Ahomia gen. nov. and novel species of Ahomia, Amazonocrinis, and Dendronalium from the biodiversity-rich northeast region of India.

Five cyanobacterial strains exhibiting Nostoc-like morphology were sampled from the biodiversity hotspots of the northeast region of India and characterized using a polyphasic approach. Molecular and phylogenetic analysis using the 16S rRNA gene indicated that the strains belonged to the genera Amazonocrinis and Dendronalium. In the present investigation, the 16S rRNA gene phylogeny clearly demarcated two separate clades of Amazonocrinis. The strain MEG8-PS clustered along with Amazonocrinis nigriterrae CENA67, which is the type strain of the genus. The other three strains ASM11-PS, RAN-4C-PS, and NP-KLS-5A-PS clustered in a different clade that was phylogenetically distinct from the Amazonocrinis sensu stricto clade. Interestingly, while the 16S rRNA gene phylogeny exhibited two separate clusters, the 16S-23S ITS region analysis did not provide strong support for the phylogenetic observation. Subsequent analyses raised questions regarding the resolving power of the 16S-23S ITS region at the genera level and the associated complexities in cyanobacterial taxonomy. Through this study, we describe a novel genus Ahomia to accommodate the members clustering outside the Amazonocrinis sensu stricto clade. In addition, we describe five novel species, Ahomia kamrupensis, Ahomia purpurea, Ahomia soli, Amazonocrinis meghalayensis, and Dendronalium spirale, in accordance with the International Code of Nomenclature for algae, fungi, and plants (ICN). Apart from further enriching the genera Amazonocrinis and Dendronalium, the current study helps to resolve the taxonomic complexities revolving around the genus Amazonocrinis and aims to attract researchers to the continued exploration of the tropical and subtropical cyanobacteria for interesting taxa and lineages.

Circumscription of Fulbrightiella gen. nov. and Sherwoodiella gen. nov., Two Novel Genera in the Calotrichaceae (Nostocales, Cyanobacteria).

Three novel strains in Calotrichaceae from tropical habitats were isolated and characterized with regard to their morphology, phylogenetic placement, and secondary structures of conserved domains in the 16S-23S internal transcribed spacer (ITS). The strains fell into two clades formerly identified as Calothrix from freshwater and brackish habitats. Based on both morphology and ecology, they differed from the type species of Calothrix, C. confervicola, which is marine, has wide trichomes with short cells, and narrows abruptly to a hyaline hair. The first clade grouped species with heteropolar filaments widened at the base and narrowed gradually toward the apex but not ending in a hair, with basal heterocytes that are formed in series as the apically placed heterocytes senesce; this clade is being named Fulbrightiella gen. nov., with two named species, F. bharadwajae sp. nov. and F. oahuensis sp. nov. The second clade was comprised of a single species with isopolar trichomes that are untapering as hormogonia, but which widen midfilament and taper toward both ends following growth. These trichomes develop pairs of heterocyte mid-filament, causing fragmentation into heteropolar trichomes with basal heterocytes and ends that taper, but not to a hair. This clade consists of a single species at present, Sherwoodiella mauiensis. With this action, four clades in the Calotrichaceae have been named: Macrochaete, Dulcicalothrix, Fulbrightiella, and Sherwoodiella. Calothrix sensu stricto is truly marine, morphologically distinct, and unsequenced; finding and sequencing the generitype for Calothrix remains as the most important and unfinished task in the revision of the Calotrichaceae.

Phycological exploration of the global biodiversity hotspots of Northeast India: discovery of a new species of soil-dwelling cyanobacteria, Desikacharya kailashaharensis sp. nov.

A soil-dwelling cyanobacterial strain (KLS-BP-3A_PS), has been isolated from the biodiversity rich Northeast region of India and characterized using a polyphasic approach. The strain was collected from a field covered with grass, near a stream from the Unakoti district of Tripura. Upon culturing in the laboratory, initial studies indicated the strain to be showing typical Nostoc or Nostoc-like morphology. Subsequently, 16S rRNA gene phylogenetic analyses using Neighbour joining, Maximum-likelihood, and Bayesian inference methods gave a distinct and stable positioning of the strain inside the genus Desikacharya. Upon recovery of the full-length operon of the 16S-23S ITS region with both tRNAs (tRNAIle and tRNAAla), the folded secondary structures revealed unique patterns of the D1-D1', V2, Box-B, and V3 regions of the strain KLS-BP-3A_PS as compared to phylogenetically related species of the genus Desikacharya. The total evidence approach indicated conclusively that the strain under investigation is a new species of the genus Desikacharya, which we describe as Desikacharya kailashaharensis in accordance with the International Code of Nomenclature for algae, fungi, and plants. Further, 16S rRNA gene phylogeny and evaluation of the 16S-23S ITS operons along with implying a re-examination of the family level affiliation of Desikacharya as well its generic limits may be in order. Notably, this study brings into focus the very less explored Northeast region of India which shares two global biodiversity hotspots in the world.

Digging deeper into the taxonomy of Cylindrospermum and description of Johanseniella tripurensis gen. et sp. nov. from India.

The wet soil-inhabiting cyanobacterium URH-6-PS was collected from the state of Tripura, India, and characterized using a polyphasic approach. Initial microscopic analysis indicated the strain to be a member of the genus Cylindrospermum, however, the 16S rRNA gene phylogenetic analysis showed some interesting results. The strain URH-6-PS clustered in the Clade I, which positioned itself outside the Cylindrospermum sensu stricto clade with strong probability/bootstrap support, indicating that the strain may not belong to the genus Cylindrospermum. Further, analysis of the 16S-23S ITS region using the folded secondary structures of the D1-D1', Box-B, and V3 helices and the 16S-23S ITS percentage dissimilarity values clearly indicated the distinctiveness of strain URH-6-PS from other members of the Clade I. The detailed investigations conducted in this study provided sufficient evidence that the taxonomic status of the members of Clade I need to be revised. Interestingly, all members of the clade I originated from tropical and subtropical habitats while members of the Cylindrospermum sensu stricto clade have been reported from temperate conditions. Previous studies had also indicated the possibility of the formation of tropical and subtropical Cylindrospermum-like genera, and thus this comes across as a much-awaited study to resolve the taxonomic complexities around the genus Cylindrospermum. Based on the results obtained in this study and the trends observed in the earlier studies, we describe a novel Cylindrospermum-like genus Johanseniella gen. nov., with the type species Johanseniellatripurensis sp. nov. in accordance with the International Code of Nomenclature for algae, fungi, and plants.

Insights into the phylogenetic inconsistencies of the genus Amazonocrinis and description of epilithic Amazonocrinis malviyae sp. nov. (Cyanobacteria, Nostocales) from Jammu and Kashmir, India.

A dark-coloured thin film of cyanobacteria growing on the bottom of a submerged stone was isolated from Basantgarh village in Udhampur district, Jammu and Kashmir, India. The isolated strain (designated 19C-PST) was characterized using a polyphasic approach. The strain exhibited typical Nostoc-like morphology with a characteristic feature of having heterocytes in series. The 16S rRNA gene phylogeny placed the strain at a well-supported and distinct node. Notably, the recently described genus, Amazonocrinis, on the addition of more 16S rRNA gene sequences, reflected a critical split, which proved to be stable and well supported in all phylogenetic analyses of the 16S rRNA gene. Interestingly, Amazonocrinis nigriterrae CENA67T (type species of the genus) clustered together with our strain 19C-PST in the 16S rRNA gene phylogenetic analysis while the rest of the members of the genus Amazonocrinis were placed at a separate and distant node. This clearly indicated that strain 19C-PST is a member of Amazonocrinis sensu stricto. However, the results of phylogenetic analysis of ITS sequences only, in strains purported to belong to Amazonocrinis did not agree with the 16S rRNA gene results and placed our strain 19C-PST in a sister clade to three strains that have not yet been speciated, UHCC 0702, NIES-4103 and SA22, with A. nigriterrae falling into a separate clade. Further, folded secondary structures of the D1-D1', V2, BoxB and V3 helices of strain 19C-PST were found to be significantly different from those of all the phylogenetically related taxa. The study revealed an interesting case where low taxon sampling and phylogenomic interpretations came across as points of attention in cyanobacterial taxonomy. Based on the morphological, phylogenetic, 16S-23S ITS secondary structure analyses, we describe our strain as Amazonocrinis malviyae sp. nov. in accordance with the International Code of Nomenclature for algae, fungi and plants. This work also illuminates the need for further research to resolve the taxonomic discrepancies among Amazonocrinis strains.

Constrictifilum karadense gen. et sp. nov., a new Nostocalean genus from Maharashtra, India.

A freshwater dwelling cyanobacterium (strain MKW3) was isolated from a sample collected from a water logged sugarcane field located in Malkapur, Karad, Maharashtra, India, and was characterized using a polyphasic approach. In the 16S rRNA gene phylogenetic analysis, strain MKW3 clustered with two misidentified strains-Nostoc sp. CENA239 and Calothrix sp. NIES2100. The phylogenetically related members included strains identified as Nostoc, Aulosira, Calothrix, Tolypothrix, Camptylonemopsis and Microchaete. The phylogenetic and the morphological analysis of the strain MKW3 indicated that it does not belong to any of the above mentioned genera. Furthermore, the 16S-23S ITS secondary structure analysis provided clear evidence indicating that strain MKW3 is different from Nostoc sp. CENA239 and Calothrix sp. NIES2100. Based on the morphological, phylogenetic and 16S-23S ITS secondary structure analysis we describe our strain as Constrictifilum karadense gen. et sp. nov. in accordance with the International Code of Nomenclature for algae, fungi and plants.

Issues in cyanobacterial taxonomy: comprehensive case study of unbranched, false branched and true branched heterocytous cyanobacteria.

The order Nostocales is represented by morphologically diverse forms with respect to the branching patterns and polarity of the filaments. With growing understanding of taxonomy and systematics, members of the order Nostocales have also undergone multiple taxonomic revisions. The last decade has seen a surge in the description of new genera and families within the order Nostocales. In this study, we discuss the taxonomic status of all the newly described and reclassified taxa of some of the prominent morphological forms within the order Nostocales by constructing comprehensive phylogenetic trees. Further, we propose certain strategies that would contribute to resolving the taxonomic complexities arising due to inadequate taxon sampling.

Fortiea necridiiformans sp. nov., a soil-dwelling cyanobacterium from Pachmarhi Biosphere Reserve, India.

Cyanobacterial strain PS4G was isolated from seepage soil sampled at Pachmarhi, Madhya Pradesh, India, and was characterized using a polyphasic approach. The results of morphological analysis showed that strain PS4G had unique morphological characteristics which were not observed in the other described species of the genus Fortiea. In the 16S rRNA gene phylogenetic analysis inferred using Bayesian inference, maximum-likelihood and neighbour-joining methods, strain PS4G clustered within the clade consisting of the members of the genus Fortiea. Furthermore, in the secondary structure analysis using the D1-D1' helix and BoxB regions of 16S-23S ITS, strain PS4G showed marked differences in comparison with other members of the genus Fortiea. Overall, the morphological, phylogenetic and folded 16S-23S ITS secondary structure examination indicated that strain PS4G represents a novel species of the genus Fortiea. In accordance with the International Code of Nomenclature of Algae, Fungi and Plants we describe a novel species of Fortiea with the name Fortiea necridiiformans sp. nov.

Description of novel species of Aliinostoc, Desikacharya and Desmonostoc using a polyphasic approach.

Five cyanobacterial strains with Nostoc-like morphology from different localities of the Mazandaran province of Iran were characterized using a polyphasic approach. Three strains clustered within the Aliinostoc clade whereas one each of the remaining two strains clustered within the genera Desmonostoc and Desikacharya. The phylogenetic positioning of all the strains by the bayesian inference, neighbour joining and maximum parsimony methods inferred using 16S rRNA gene indicated them to represent novel species of the genera Aliinostoc, Desmonostoc and Desikacharya. The 16S-23S ITS secondary structure analysis revealed that all five strains under study represented novel species unknown to science. In accordance with the International Code of Nomenclature for algae, fungi and plants we describe three novel species of the genus Aliinostoc and one species each of the genera Desmonostoc and Desikacharya.

Phylogenetic evaluation of the genus Nostoc and description of Nostoc neudorfense sp. nov., from the Czech Republic.

Cyanobacterial strain ARC8 was isolated from seepage coming into the river Dracice, Frantiskov, Czech Republic, and was characterized using a polyphasic approach. Strain ARC8 showed a typical Nostoc-like morphology and in-depth morphological characterization indicated that it is a member of the genus Nostoc. Furthermore, in the 16S rRNA gene phylogeny inferred using Bayesian inference, maximum likelihood and neighbour joining methods, strain ARC8 clustered within the Nostoc sensu stricto clade. The phylogenetic distance and the positioning of strain ARC8 also indicated that it is a member of the genus Nostoc. Furthermore, the rbcL gene phylogeny along with the 16S-23S ITS secondary structure analysis also supported the findings from the 16S rRNA gene tree. In accordance with the International Code of Nomenclature for Algae, Fungi and Plants we describe a novel species of Nostoc with the name Nostoc neudorfense sp. nov.

Phylogenetic evaluation of the true-branched heterocytous cyanobacteria and description of soil dwelling Westiellopsis akinetica sp. nov.

Soil inhabiting true branching heterocytous cyanobacterium strain SNS 3 with T-type branching was collected from the campus of Banaras Hindu University, India and characterized using the polyphasic approach. Morphological observations showed the presence of akinetes (in chain), hormogonia and monocytes. Physiological characterization of strain SNS 3 showed high content of carotenoid in comparison to chlorophyll a content along with also exhibiting a higher C/N ratio in the nitrogen deficient BG110 medium. 16S rRNA gene sequencing and subsequent phylogeny indicated strong clustering of the strain SNS 3 within the Westiellopsis clade. Folded secondary structures of the ITS region showed visible differences in the D1-D1' and BoxB helix of the strain SNS 3. The polyphasic approach indicated the strain SNS 3 as a new member of the genus Westiellopsis with the name proposed being Westiellopsis akinetica sp. nov. on the basis of the International Code of Nomenclature for Algae, Fungi and Plants. Intense phylogenetic evaluation of the entire true-branched heterocytous clade indicated the need for more revisionary attempts for demarcating the sensu stricto clades along with highlighting the scope for further taxonomic revisions in the future.

Phylogenetic complexities of the members of Rivulariaceae with the re-creation of the family Calotrichaceae and description of Dulcicalothrix necridiiformans gen nov., sp nov., and reclassification of Calothrix desertica.

A freshwater dwelling, tapering, heterocytous cyanobacterium (strain V13) was isolated from an oligotrophic pond in the Shrirampur taluka, Ahmednagar district of Maharashtra in India. Initial morphological examination indicated that strain V13 belonged to the genus Calothrix. Subsequent molecular and phylogenetic assessment based on 16S rRNA gene, led us to describe the freshwater/terrestrial clade of Calothrix strains without terminal hairs as a new genus Dulcicalothrix gen. nov., with the type species Dulcicalothrix necridiiformans sp. nov. (Strain V13) on the basis of the necridia forming ability of the strain. Also, the 16S-23S ITS secondary structure analysis clearly differentiated strain V13 from the other members of the clade. Past studies and the current state of knowledge makes it imperative to separate the groups Calothrix (marine/freshwater Calothrix), Macrochaete and Dulcicalothrix (freshwater/terrestrial Calothrix) into separate genera in accordance with the International Code of Nomenclature for Algae, Fungi and Plants. Robust phylogenetic evidence and previous reports strongly support the re-erection of the family Calotrichaceae distinct from the existing family Rivulariaceae.

Desikacharya gen. nov., a phylogenetically distinct genus of Cyanobacteria along with the description of two new species, Desikacharya nostocoides sp. nov. and Desikacharya soli sp. nov., and reclassification of Nostoc thermotolerans to Desikacharya thermotolerans comb. nov.

Two Nostoc-like strains have been isolated, purified, cultured and identified on the basis of the polyphasic approach using morphological, ecological, molecular and phylogenetic methods. Both strains were found to have morphology similar to the genus Nostoc, but clustered strongly in a group distant from the Nostocsensu stricto clade. Further analysis, using the folded structures of the 16S-23S ITS region revealed strong differences from closely related members of the genus Nostoc. Distinct phylogenetic clustering and strong tree topologies using Bayesian inference, maximum-likelihood and maximum-parsimony methods indicated the need to revisit the taxonomy of the members of this particular clade with a clear need for giving a generic status distinct from the genus Nostoc. In accordance with the International Code of Nomenclature for Algae, Fungi and Plants, the name Desikacharya gen. nov. is proposed for the new genus along with the description of two new species, Desikacharya nostocoides sp. nov. and Desikacharya soli sp. nov., and reclassification of Nostoc thermotolerans to Desikacharya thermotolerans comb. nov.

Description of two new species of Aliinostoc and one new species of Desmonostoc from India based on the Polyphasic Approach and reclassification of Nostoc punensis to Desmonostoc punense comb. nov.

Three heterocytous cyanobacterial strains were isolated from different habitats of Central India, and initial morphological studies indicated them to be members of the genus Nostoc or other closely related genera. Subsequent studies using morphological, ecological, molecular and phylogenetic methods indicated the three strains to be new members of the genera Aliinostoc and Desmonostoc. Folding of the D1-D1' helix of the ITS region clearly differentiated the three strains from the other closely related strains, thus providing final indications of the strains being different and new additions to the genera Aliinostoc and Desmonostoc. In accordance with the International Code of Nomenclature for algae, fungi and plants, we establish three new species: Aliinostoc tiwarii sp. nov, Aliinostoc soli sp. nov. and Desmonostoc magnisporum sp. nov. along with reclassifying Nostoc punensis as Desmonostoc punense comb. nov.

Insights into the phylogeny of false-branching heterocytous cyanobacteria with the description of Scytonema pachmarhiense sp. nov. isolated from Pachmarhi Biosphere Reserve, India.

A false branching cyanobacterium (strain 10A1_PS) was isolated from a freshwater body of the Pachmarhi Biosphere Reserve, India and was characterised using the polyphasic approach. The detailed morphological examination indicated that the strain belonged to the complex genus Scytonema as it exhibited typical false branching character whose frequency increased with age of the culture. As the family Scytonemataceae and the genus Scytonema has been shown to be polyphyletic in many studies, we provide deep insights into the phylogenetic complexities within the family Scytonemataceae based on 16S rRNA gene phylogeny along with complete morphological, molecular and phylogenetic characterisation of the strain. The 16S rRNA gene phylogenetic tree inferred by Bayesian Inference, Neighbor-Joining and Maximum Parsimony methods showed that the strain clustered within the Scytonema sensu stricto clade. The phylogenetic distance and the positioning of the strain clearly indicated it to be different from other Scytonema species. Further analysis using rbcL phylogeny, folded secondary structures of the 16S-23S ITS, p-distance and percentage pairwise similarity matrix clearly distinguished the strain 10A1_PS from the other closely related species. In accordance with the International Code of Nomenclature for Algae, Fungi and Plants, we propose the name of the new species to be Scytonema pachmarhiense.

Nostoc thermotolerans sp. nov., a soil-dwelling species of Nostoc (Cyanobacteria).

A filamentous, soil-dwelling cyanobacterial strain (9C-PST) was isolated from Mandsaur, Madhya Pradesh, India, and is described as a new species of the genus Nostoc. Extensive morphological and molecular characterization along with a thorough assessment of ecology was performed. The style of filament orientation, type and nature of the sheath (e.g. distribution and visibility across the trichome), and vegetative and heterocyte cell dimensions and shape were assessed for over one year using both the laboratory grown culture and the naturally occurring samples. Sequencing of the 16S rRNA gene showed 94 % similarity with Nostocpiscinale CENA21 while analyses of the secondary structures of the 16S-23S ITS region showed unique folding patterns that differentiated this strain from other species of Nostoc. The level of rbcl and rpoC1 gene sequence similarity was 91 and 94 % to Nostocsp. PCC 7524 and Nostocpiscinale CENA21, respectively, while the nifD gene sequence similarity was found to be 99 % with Nostocpiscinale CENA21. The phenotypic, ecological, genetic and phylogenetic observations indicate that the strain 9C-PST represents a novel species of the genus Nostoc with the name proposed being Nostoc thermotolerans sp. nov. according to the International Code of Nomenclature for Algae, Fungi, and Plants.