ABSTRACT
Two new species of Dulcicalothrix, D. adhikaryi sp. nov. and D. iyengarii sp. nov., were discovered in India and are characterized and described in accordance with the rules of the International Code of Nomenclature for algae, fungi, and plants (ICN). As a result of phylogenetic analysis, Calothrix elsteri is reassigned to Brunnivagina gen. nov. During comparison with all Dulcicalothrix for which sequence data were available, we observed that the genus has six ribosomal operons in three orthologous types. Each of the three orthologs could be identified based upon indels occurring in the D1-D1' helix sequence in the ITS rRNA region between the 16S and 23S rRNA genes, and in these three types, there were operons containing ITS rRNA regions with and without tRNA genes. Examination of complete genomes in Dulcicalothrix revealed that, at least in the three strains for which complete genomes are available, there are five ribosomal operons, two with tRNA genes and three with no tRNA genes in the ITS rRNA region. Internal transcribed spacer rRNA regions have been consistently used to differentiate species, both on the basis of secondary structure and percent dissimilarity. Our findings call into question the use of ITS rRNA regions to differentiate species in the absence of efforts to obtain multiple operons of the ITS rRNA region through cloning or targeted PCR amplicons. The ITS rRNA region data for Dulcicalothrix is woefully incomplete, but we provide herein a means for dealing with incomplete data using the polyphasic approach to analyze diverse molecular character sets. Caution is urged in using ITS rRNA data, but a way forward through the complexity is also proposed.
ABSTRACT
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.
Subject(s)
Exploratory Behavior , Nostoc , Phylogeny , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , Bacterial Typing Techniques , Sequence Analysis, DNA , Nostoc/genetics , Biodiversity , IndiaABSTRACT
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.
Subject(s)
Cyanobacteria , Ecosystem , Phylogeny , RNA, Ribosomal, 16S , DNA, Bacterial , Cyanobacteria/genetics , Sequence Analysis, DNAABSTRACT
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.
Subject(s)
Nostoc , Soil , RNA, Ribosomal, 16S/genetics , Phylogeny , DNA, Bacterial/genetics , Bayes Theorem , Bacterial Typing Techniques , Sequence Analysis, DNA , Nostoc/genetics , BiodiversityABSTRACT
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.
Subject(s)
Cyanobacteria , Bacterial Typing Techniques , Cyanobacteria/genetics , DNA, Bacterial/genetics , Fatty Acids , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
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.
Subject(s)
Fatty Acids , Nostoc , Phylogeny , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , Sequence Analysis, DNA , Bacterial Typing Techniques , Base Composition , Fatty Acids/chemistry , IndiaABSTRACT
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.