| Home | E-Submission | Sitemap | Contact Us |  
top_img
Algae > Volume 39(2); 2024 > Article
Nuryadi, Sumimoto, and Suda: Discovery of novel Nodosilinea species (Cyanobacteria, Nodosilineales) isolated from terrestrial habitat in Ryukyus campus, Okinawa, Japan

ABSTRACT

Terrestrial cyanobacteria are extremely diverse. In urban areas, they can be found as black stains on the surface of building walls, stone monuments, or man-made structures. Many of the terrestrial cyanobacteria are still understudied. To expand knowledge of terrestrial cyanobacterial diversity, a polyphasic characterization was performed to identify 12 strains isolated from campus of University of the Ryukyus, Okinawa, Japan. Multigene phylogenetic analyses based on 16S rRNA gene and 16S-23S rRNA internal transcribed spacer (ITS) region showed that the isolated strains formed two independent subclades within Nodosilinea, and were distantly related to all described Nodosilinea species. The 16S-23S rRNA ITS secondary structures showed variations for D1-D1′ and Box B domain, while V3 domain was almost identical among entire species of Nodosilinea, including the studied strains. In addition, a unique morphological character, i.e. forming nodule or spiral shape, was also observed in certain studied strains. According to polyphasic characterization, Nodosilinea coculeatus sp. nov. and Nodosilinea terrestrialis sp. nov., were proposed as two new species of terrestrial cyanobacteria from Okinawa.

INTRODUCTION

Cyanobacteria are among the most geographically widespread oxygenic photosynthetic organisms (Whitton and Potts 2000). They can be found in various habitats, such as terrestrial, freshwater, marine, or even in extreme environments (Dvořák et al. 2015, Sciuto and Moro 2016, Kim et al. 2022). Cyanobacteria play crucial ecological roles in providing carbon, oxygen, and nitrogen fixers in global ecosystems (Charpy et al. 2010, 2012, Gaydon et al. 2012, Brocke et al. 2018). Despite their wide distributions and essential ecological contributions, taxonomic classification of cyanobacteria is extremely complex. Numerous cryptic taxa have been incorrectly classified. Traditional identification, solely based on morphology without considering genetic differences has led to taxonomic confusion because of the similarity in morphological characteristics among different taxa (Komárek 2018). It has been reported that the morphological classification does not accurately correspond to the actual evolutionary history of cyanobacteria (Zammit et al. 2012, Mühlsteinová et al. 2018). Recently, integration of traditional morphological and modern molecular identification, known as polyphasic characterization, has emerged as the most effective approach to correctly classifying cyanobacterial taxa. It has now become fundamental framework for taxonomic classification of cyanobacteria (Hoffmann et al. 2005, Komárek 2016, Caires et al. 2018).
In the last century, cyanobacterial taxonomic classification has massively changed (Komárek et al. 2014). The most recent taxonomic revision, based on multigene phylogeny by Strunecký et al. (2023), proposed ten new orders and 15 new families to achieve monophyly in all taxonomic ranks. One of the newly proposed orders is Nodosilineales. This order consists of filamentous cyanobacteria with isodiametric or two times longer than wider cells, very thin cell width (<2 μm), and parietal thylakoids. Due to lack of morphologically diagnostic features, many strains of Nodosilineales were incorrectly assigned to Leptolyngbya sensu stricto, which is one of the most problematic genera in cyanobacteria. The most diverse genus in Nodosilineales is Nodosilinea, containing at least 10 described species (Perkenson et al. 2011, Heidari et al. 2018, Vázques-Martínez et al. 2018, Radzi et al. 2019, Davydov et al. 2020, Cai et al. 2022). Nodosilinea species have capability to form nodules in their filaments under low light conditions, a unique morphological character that can be used to distinguish them from Leptolyngbya species and other species in the Pseudanabaenaceae (Perkenson et al. 2011). Despite numerous taxonomic studies on Nodosilinea, some species of Nodosilinea remain undescribed. Therefore, increased surveys from unexplored regions are still necessary to uncover the true diversity of this genus.
Okinawa is located in a subtropical region influenced by high temperature, humidity, and intense sunlight exposure. This climate condition can promote proliferation of terrestrial microbial biofilms (Hoffmann 1989, Lewin 2006) which are usually found in residential or public areas, forming blackened stains on the surface of old building, stone monuments, or other man-made structures. The current main campus of Ryukyus University located in Nishihara, Okinawa has been utilized since 1975. Nowadays, various types of microbial biofilms have grown in the campus areas, including on building walls, the university monument, or limestone stairs. These microbial biofilms constitute a complex ecosystem, harboring extremely diverse microorganisms, including subaerial cyanobacteria. Surprisingly, the diversity of the cyanobacteria from Ryukyus campus is extraordinarily diverse, however, the specific status of many strains is still undescribed (Nguyen et al. 2017). Based on this information, we conducted more detailed identification for the undescribed cyanobacterial species from Nishihara campus of University of the Ryukyus. As the result, two potential novel species from genus Nodosilinea (order Nodosilineales) have been discovered. Furthermore, two new species name, Nodosilinea coculeatus sp. nov. and Nodosilinea terrestrialis sp. nov., are proposed as new members of Nodosilinea under the provision of the International Code of Nomenclature for Algae, Fungi and Plants (ICN).

MATERIALS AND METHODS

Culture conditions

A total 12 terrestrial cyanobacterial strains from our culture collections were analyzed for the taxonomic identification of Nodosilinea species. The strains were previously isolated from surfaces of building walls, stairs, monuments, or roadside walls within Ryukyus campus (Nguyen et al. 2017). All strains were maintained using BG-11 with pH 7.4, and kept under 14 : 10 (L : D) cycle using light intensity of approximately 40 μE m−2 s−1 at 25 ± 1°C. In addition, experiment using a nitrogen-depleted medium was performed to observe nodule formation in all isolated strains. Nodosilinea strains were examined under the light microscope after four weeks incubation.

DNA extraction and polymerase chain reaction amplifications

Genomic DNA was extracted using DNeasy plant mini kit (Qiagen, Hilden, Germany) following manufacturer’s protocol. Two genetic markers, 16S rRNA gene and 16S-23S rRNA internal transcribed spacer (ITS), were amplified for molecular identification of the targeted strains. Two pairs of primers, 27F (Neilan et al. 1997) with CYA1371R(1+2+3) (Murakami et al. 2004) and CYA781F (Nübel et al. 1997) with 23S30R (Taton et al. 2003), were used to amplify 16S rRNA gene and 16S-23S ITS region, respectively. The amplification of 16S rRNA and 16S-23S rRNA ITS region was performed following protocols outlined in Nuryadi and Suda (2022). Subsequently, the quality of polymerase chain reaction products was checked using 1% agarose gel. The amplicons with a single bright DNA band were sent to Macrogen Japan Corp. (Tokyo, Japan) for sequencing process in forward and reverse directions using the same pairs of primers for amplification.

Molecular analyses of the 16S rRNA gene and the 16S-23S ITS region

Phylogenetic analyses were performed based on sequences of 16S rRNA gene and 16S-23S rRNA ITS region. Sequences of studied strains were aligned with sequences of various cyanobacterial species obtained from GenBank, including Nodosilinea nodulosa UTEX 2910 (EF122600) as the type species of Nodosilinea. The alignment was performed separately for each gene using MUSCLE algorithm (Edgar 2004) in MEGA X software (Kumar et al. 2018), then, the alignment result was manually corrected. Both phylogenetic trees, 16S rRNA gene and 16S-23S rRNA ITS region, of Nodosilinea and related taxa were constructed using both maximum likelihood (ML) and Bayesian inference (BI). Appropriate substitution models (Table 1) were separately selected for each gene using jModelTest 2.1.6 (Darriba et al. 2012, Guindon and Gascuel 2023) through CIPRES science gateway (Miller et al. 2010). The ML phylogenetic tree was constructed using RaxMLGUI 2.0 (Edler et al. 2021) with 1,000 bootstrap replications. The BI analysis was performed using MrBayes 3.2.7a (Ronquist and Huelsenbeck 2003) through CIPRES science gateway, with four Markov chain Monte Carlo simulations, 107 generations, resampling every 100 generations, and 0.25 burnin rate. A similar setting was applied for 16S rRNA genes and 16S-23S rRNA ITS region. A matrix of genetic similarity among all Nodosilinea species, including strains from Okinawa, was estimated based on calculation of p-distance in MEGA X.
The 16S-23S rRNA ITS secondary structures of each strain were constructed using Mfold version 2.3 with default parameters (Zucker 2003). The identifiable secondary structures were compared between studied strains with described species of Nodosilinea. All predicted structures of Nodosilinea species were identified following Iteman et al. (2000), Perkerson et al. (2011), Heidari et al. (2018), Vázques-Martínez et al. (2018), Radzi et al. (2019), Davydov et al. (2020), and Cai et al. (2022). All sequences of Nodosilinea strains from Okinawa were deposited to GenBank under the accession numbers LC731253–LC731264.

Morphological observation

Morphological characters were observed using an inverted microscope (CKX31; Olympus, Tokyo, Japan) and light microscope (Nikon Eclipse 80i; Nikon, Tokyo, Japan) equipped with Spot Idea 5 MP camera (Diagnostic Instrument, Sterling Heights, MI, USA). Morphological features such as filament shape, sheaths, cell shape, cell dimensions, cross-walls, apical cells, and nodule formation were observed from a total 50 filaments for morphological characterization. Subsequently, all isolated strains were morphologically identified according to Komárek and Anagnostidis (2005), Perkerson et al. (2011), Heidari et al. (2018), Vázques-Martínez et al. (2018), Radzi et al. (2019), Davydov et al. (2020), and Cai et al. (2022).

RESULTS

Phylogenetic analyses

Both ML and BI phylogenetic trees demonstrated almost similar topologies. The ML phylogenetic tree was used to describe phylogenetic relationship among Nodosilinea with other related cyanobacterial taxa. Based on the 16S rRNA phylogeny, two new species, Nodosilinea coculeatus H. Nuryadi, S. Sumimoto and S. Suda and Nodosilinea terrestrialis H. Nuryadi, S. Sumimoto and S. Suda, were described from 12 strains isolated from terrestrial habitat within Ryukyus University areas. Each new species formed highly supported monophyletic subclade within Nodosilinea clade, and they were genetically distant to other described species (Fig. 1). According to the 16S rRNA gene phylogenetic inference, the closest related species to N. coculeatus and N. terrestrialis were N. signiensis and N. bijugata, respectively. The genetic similarity between strains of N. coculeatus and N. signiensis were 99.1–99.2%, whereas strains of N. terrestrialis shared 98.8% similarity to N. bijugata (Table 2). Strains of N. coculeatus shared 99.9–100% similarity one another, and N. terrestrialis was 100% identical among all strains. N. coculeatus shared 97.7–97.8% similarity to N. terrestrialis. The 16S-23S rRNA ITS also showed that both N. coculeatus and N. terrestrialis were clearly distinct from other described species of Nodosilinea (Fig. 2). All strains of N. coculeatus were sister to Nodosilinea sp. IkpPStn44 with a genetic similarity of 93%, while N. terrestrialis shared a 96.5% genetic similarity to N. bijugata (Table 3).

16S-23S rRNA ITS secondary structures

Three informative domains of the 16S-23S rRNA ITS region: D1-D1′, Box B, and V3 were successfully identified in 12 strains from Okinawa. The structure length was nearly consistent for each structure, except for Box B domain (Table 4). The structure of D1-D1′ was composed of 61 to 62 bp of nucleotide sequences. Despite having almost similar length, this domain exhibited variation in structure form among species (Fig. 3). The bases of the D1-D1′ structures of every species had almost identical shape. They were characterized by a conserved 5 bp of basal stem (5′-GACCU–AGGUC-3′) followed by 6 bp of unilateral bulge in 3′ side. Two 1 bp unilateral bulges were found on the 3′ and 5′ side in long stem helix. Some variations in nucleotide sequences were identified in this long stem helix. A distinct shape was clearly observed in the top of structure. Three species: N. coculeatus, N. ramsarensis, and Nodosilinea sp. IkpPStn44 had a 1 : 4, 1 : 3, and 1 : 3 bilateral bulge, respectively, followed by 4 bp terminal loop with a short stem at the top of structure. One species, N. epilithica Kovacik 1998/7, presented a 1 : 1 bilateral bulge before 10 bp of terminal loop. The remaining species, including N. terrestrialis showed a 16 bp of terminal loop with slight differences in nucleotide composition. Furthermore, both tRNAIle and tRNAAla were present in all species of Nodosilinea (secondary structures were not shown), while structure of V2 domain was absent in all species of Nodosilinea.
In contrast to D1-D1′ domain, variations in length and shape were observed in the Box B domain in some species of Nodosilinea (Fig. 4). The length of Box B structure ranged from 39 to 47 bp. In terms of structure shape, all species shared similar conserved 4 bp of basal stem (5′-AGCA–UGCU-3′), followed by varied structure shapes in certain species. N. coculeatus and N. nodulosa had two unilateral bulges 5′ and 3′ side before 4 bp terminal loop with a long stem. Another new species, N. terrestrialis was almost identical with N. bijugata. The structure showed two bilateral bulges after conserved basal stem. However, 4 and 5 bp of terminal loops were found in N. terrestrialis and N. bijugata, respectively. The V3 domain showed the most conserved structure in every species of Nodosilinea (Fig. 5). All species had 5 bp of basal stem with nucleotide substitution U and C in fifth bases (5′-CAUUU/C–GAGUG-3′). A 4 : 2 bilateral bulge followed by a 4 or 5 bp of terminal loop with a 3 bp of stem was identified in the top of structure.

Morphological identification

A total 12 strains were isolated from several sampling sites in the university area. The filaments of 12 strains comprising from two species, N. coculeatus (Fig. 6) and N. terrestrialis (Fig. 7), exhibited identical morphological characteristics. In general, all strains were characterized as wavy filaments, blue-green coloration, with almost isodiametric cells or mostly longer than wide, distantly constricted at the cross-walls, rounded apical cells without calyptra and enclosed with thin colorless sheaths. Under a nitrogen-depleted condition, filaments of N. coculeatus formed spiral shape (Fig. 6E & F), while N. terrestrialis showed presence of nodule formation (Fig. 7E & F). No nodule or spiral shape were observed in the filaments of two new species cultured in medium containing nitrogen source (Figs 6C & 7D). Moreover, heterocytes or other specialized cells were not observed in any of the studied strains. The morphological characteristics of all species of Nodosilinea, including new species from Okinawa, are presented in Table 5.

Nodosilinea coculeatus H. Nuryadi, S. Sumimoto et S. Suda sp. nov

Description

Thallus forms blue-green fasciculate, thin layer mats in culture conditions. Filament typically very thin, straight or sometimes curvy. Sheath generally thin, colorless. Trichome blue-green, composed from barrel shaped-cells or sometimes almost isodiametric. Cell dimension usually longer (1.4–2.7 μm) than wide (1.2–1.5 μm), cells well separated by cross-walls, not granulated, heterocytes or other specialized cells absent. Apical cells rounded without calyptra. Forming spiral shape in nitrogen-depleted condition. Forming hormogonia by straight trichome fragmentation without necridic cell in reproduction process.

Holotype

A metabolically inactive dried specimen from reference strain Ru1-11, RYU A0060 in the RYU (Herbarium of the Faculty of Science, University of the Ryukyus, Okinawa, Japan) sampled from building walls at campus of University of the Ryukyus, Nishihara, Okinawa, Japan.

Reference strain

Nodosilinea coculeatus Ru1-11 (GenBank accession No. LC731255).

Etymology

coculeatus from Latin meaning “spiral”, referring to the spiral shape which often found from filaments of this species.

Diagnosis

Forming blue-green fasciculate thallus in laboratory condition. Capable to form spiral shape in their filaments. Species delimitation based on phylogeny of 16S rRNA gene and 16S-23S rRNA ITS region, as well as distinct ITS secondary structures of D1-D1′ and Box B domains.

Habitat

Microbial biofilms growing on the surface of building walls.

Nodosilinea terrestrialis H. Nuryadi, S. Sumimoto et S. Suda sp. nov

Description

Thallus forms green to dark green fasciculate, thin mats in culture conditions. Filament typically very thin, straight sometimes curvy. Sheath generally thin, colorless. Trichome pale green, cells often almost isodiametric 1.2–2.2 μm width and 1.2–1.7 μm long, cells completely separated by ungranulated cross-walls, lacking heterocytes or other specialized cells. Apical cells rounded without calyptra. Forming nodules in poor nitrogen condition. Hormogonia formed by straight fragmentations with no formation of necridia.

Holotype

A metabolically inactive dried specimen from reference strain Ru4-25, RYU A0066 in the RYU (Herbarium of the Faculty of Science, University of the Ryukyus, Okinawa, Japan) sampled from man-made structure at campus of University of the Ryukyus, Nishihara, Okinawa, Japan.

Reference strain

Nodosilinea terrestrialis Ru4-25 (GenBank accession No. LC731263).

Etymology

terrestrialis refers to habitat of the species.

Diagnosis

Forming green to dark green fasciculate thallus in laboratory condition. Capable to form nodules in the filaments. Species delimitation based on phylogeny of 16S rRNA gene and 16S-23S rRNA ITS region, as well as distinct ITS secondary structures of D1-D1′ and Box B domains.

Habitat

Forming blackened microbial biofilms on the surface of man-made structure.

DISCUSSION

Nodosilinea has been reclassified under the newly described order Nodosilineales Strunecký and Mareš ordo.nov. and family Nodosilineaceae Strunecký and Mareš fam. nov. (Strunecký et al. 2023). This genus has relatively high species diversity, consisting at least ten described species from various environments. However, many of the taxa within this genus are still undescribed or incorrectly classified. In this study, we have described two new species of Nodosilinea, N. coculeatus and N. terrestrialis. The 16S rRNA phylogeny demonstrated that clades of Okinawan strains were genetically distant from other described Nodosilinea species. The new species, N. terrestrialis, was a sister group of N. bijugata. However, sequences of these two species were not completely identical, and they do not cluster within a single clade. Therefore, N. terrestrialis can be described as different species from N. bijugata. The genetic similarity of 16S rRNA gene sequences between new species and closely related species was >98.7% (dissimilarity <1.3%), which does not satisfy the threshold criteria for differentiating bacterial species as suggested by Stackebrandt and Ebers (2006) as well as Yarza et al. (2014). However, it is suggested that sequences of 16S rRNA gene are often insufficient for species delimitation in cyanobacteria (Bohunická et al. 2015, Bravakos et al. 2016, Akagha et al. 2023, Jusko and Johansen 2023). Recently, more variable region, such as 16S-23S rRNA ITS region, is considered as reliable genetic marker for identifying cyanobacterial species (Boyer et al. 2001, Perkerson et al. 2011, Martins and Branco 2016, Mai et al. 2018, Vázquez-Martínez et al. 2018, Martins et al. 2019, Pietrasiak et al. 2019, Lefler et al. 2021, Berthold et al. 2022). Therefore, analysis of 16S-23S rRNA ITS region is required to support speciation in cyanobacteria.
The phylogeny of 16S-23S rRNA ITS region also demonstrated genetic differences between Okinawa strains and described species of Nodosilinea. The genetic similarity was lower than 97% (dissimilarity >3%), which has exceeded criterion species differentiation established by González-Resendiz et al. (2019). In addition to phylogeny, secondary structures of informative domains in 16S-23S rRNA ITS region were constructed to provide more robust evidence for speciation. The secondary structures of 16S-23S rRNA ITS region have been widely considered as important characters for cyanobacterial taxonomic classification (Gugger et al. 2005, Johansen and Casamatta 2005, Casamatta et al. 2006), especially for describing both new genus and species (Řeháková et al. 2007, Siegesmund et al. 2008, Johansen et al. 2011, Mühlsteinová et al. 2014, Vaz et al. 2015, Brito et al. 2017, Nuryadi and Suda 2022). Despite having similar length of sequence, D1-D1′ domain exhibited variation in the structure form. Furthermore, some nucleotide substitutions can be found within identical structure. The Box B domain also exhibited diverse structure among species, both in term of sequence length and structure form. In contrast, V3 domain presented a nearly identical structure in all species. Considering variation in the secondary structure configuration, the D1-D1′ and Box B domains are appropriate for identifying two new species from Okinawa. Overall, variations in secondary structures may reflect species divergence within this genus.
Morphologically, it is very difficult to differentiate two new species with phylogenetically closely related species because they have very identical morphological features. However, they are ecologically found from different environments. Two new species are isolated from terrestrial area in subtropical region, while N. bijugata and N. signiensis are found in freshwater habitat (Perkenson et al. 2011) and extremely cold environment in Antarctica (Radzi et al. 2019), respectively. In general, the two new species from Okinawa morphologically resemble Leptolyngbya species. The morphological features frequently overlap with many species of Leptolyngbya. For instance, N. coculeatus and N. terrestrialis share similar morphological characteristics with Leptolyngbya ectocarpi (Gomont) Anagnostidis et Komárek 1988, however, the trichome color differs between them. L. ectocarpi has pinkish to pale purple trichomes, whereas both new species have blue-green trichomes. Nodule or spiral shape formation can be found in the filaments of new species but is absent in L. ectocarpi. Moreover, L. ectocarpi is a marine species, while the new species from Okinawa are found from terrestrial habitats. The morphological and ecological differences can serve as diagnostic traits, supporting molecular genetic results in the classification of N. coculeatus and N. terrestrialis as new species of cyanobacteria.
In this study, combination of some analyses, including morphology, multigene phylogeny of 16S rRNA and 16S-23S rRNA ITS region, genetic similarity, and secondary structure identification, has been applied to identify terrestrial cyanobacterial strains from Okinawa. The results of these analyses have provided compelling evidence for the description of two novel species, N. coculeatus and N. terrestrialis. The discovery of these new species has revealed high species diversity in genus Nodosilinea. Some Nodosilinea taxa are still classified as undescribed species. Further studies are necessary to define specific status of these taxa. Moreover, many locations outside campus areas have yet to be explored for terrestrial cyanobacteria. A broad exploration in various environments can lead to the discovery of new cyanobacterial species shortly.

ACKNOWLEDGEMENTS

Part of this work was supported by JSPS KAKENHI Grant (#19K06090).

CONFLICTS OF INTEREST

The authors declare that they have no potential conflicts of interest.

Fig. 1
The 16S rRNA maximum likelihood phylogenetic relationship of Nodosilinea spp. with other related cyanobacterial taxa. Two new species from terrestrial habitat in Ryukyus campus Okinawa are highlighted in gray. Numbers at the nodes indicate Bayesian posterior probabilities / bootstrap values of maximum likelihood. Only support values >0.50 and >50% are shown for posterior probabilities and bootstrap values, respectively.
algae-2024-39-2-59f1.jpg
Fig. 2
Phylogenetic inferences based on the 16S-23S internal transcribed spacer region sequences of Nodosilinea spp. Support values on the nodes are shown as Bayesian inference / maximum likelihood. Posterior probabilities >0.5 and bootstrap values >50% are shown. Gray highlight indicates new species of Nodosilinea isolated from terrestrial habitat in Ryukyus campus, Okinawa.
algae-2024-39-2-59f2.jpg
Fig. 3
Structures of D1-D1′ domain of Nodosilinea spp., including strains isolated from Okinawa. Red and blue color indicate nucleotide sequence variation in identical structures.
algae-2024-39-2-59f3.jpg
Fig. 4
Structures of Box B domain among species of Nodosilinea. Red and blue color are variable nucleotide sequences between two identical structures.
algae-2024-39-2-59f4.jpg
Fig. 5
Nearly identical structures of V3 domain among strains of Nodosilinea. Variation in nucleotide sequences is in red colors.
algae-2024-39-2-59f5.jpg
Fig. 6
Morphological characteristics of Nodosilinea coculeatus. (A) Culturable strains of N. coculeatus forming blue-green fasciculate, thin layer mats in laboratory conditions. (B & C) Filament appearances of N. coculeatus with different magnifications. (D) A colorless thin sheath in filaments of N. coculeatus (arrow). (E & F) Filaments of N. coculeatus formed spiral shapes in low nitrogen condition (arrows). Scale bars represent: B–F, 20 μm.
algae-2024-39-2-59f6.jpg
Fig. 7
Morphological appearances of Nodosilinea terrestrialis. (A) Green to dark green fasciculate, thin mats of N. terrestrialis in laboratory conditions. (B & C) Filaments characteristics observed in N. terrestrialis with different magnifications. (D) Trichome of N. terrestrialis enclosed with thin and colorless sheath (arrow). (E & F) Nodules formed by filaments of N. terrestrialis under nitrogen-depleted condition (arrows). Scale bars represent: B–F, 20 μm.
algae-2024-39-2-59f7.jpg
Table 1
Best-fit models for nucleotide substitution used in phylogenetic analyses
Gene ML BI
16S rRNA GTR + G + I GTR + I + G
16S-23S rRNA ITS region TPM2uf + I + G TPM2uf + I + G

ML, maximum likelihood; BI, Bayesian inference; ITS, internal transcribed spacer.

Table 2
Genetic similarity of Nodosilinea species based on 16S rRNA sequences
Strain 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
1 N. coculeatus Ru1-4A
2 N. coculeatus Ru1-8 100.0
3 N. coculeatus Ru1-18 99.9 99.9
4 N. coculeatus Ru1-19 100.0 100.0 99.9
5 N. coculeatus Ru1-11 100.0 100.0 99.9 100.0
6 N. coculeatus Ru1-11-17 100.0 100.0 99.9 100.0 100.0
7 N. coculeatus Ru1-11-20 100.0 100.0 99.9 100.0 100.0 100.0
8 N. terrestrialis Ru4-3 97.8 97.8 97.7 97.8 97.8 97.8 97.8
9 N. terrestrialis Ru4-7 97.8 97.8 97.7 97.8 97.8 97.8 97.8 100.0
10 N. terrestrialis Ru4-17 97.8 97.8 97.7 97.8 97.8 97.8 97.8 100.0 100.0
11 N. terrestrialis Ru4-25 97.8 97.8 97.7 97.8 97.8 97.8 97.8 100.0 100.0 100.0
12 N. terrestrialis Ru4-26 97.8 97.8 97.7 97.8 97.8 97.8 97.8 100.0 100.0 100.0 100.0
13 N. hunanesis ZJJ01 ON074585 98.2 98.2 98.1 98.2 98.2 98.2 98.2 97.0 97.0 97.0 97.0 97.0
14 N. signiensis USMFM MN585775 99.2 99.2 99.1 99.2 99.2 99.2 99.2 97.9 97.9 97.9 97.9 97.9 98.2
15 N. chupicuarensis PC471 KX859298 98.6 98.6 98.5 98.6 98.6 98.6 98.6 98.0 98.0 98.0 98.0 98.0 97.6 98.5
16 Nodosilinea sp. WD-4-2 KY098845 97.9 97.9 97.8 97.9 97.9 97.9 97.9 98.6 98.6 98.6 98.6 98.6 97.0 97.7 97.9
17 N. epilithica ACSSI 169 KY283067 97.8 97.8 97.7 97.8 97.8 97.8 97.8 98.5 98.5 98.5 98.5 98.5 96.9 97.8 97.8 99.9
18 N. bijugata KOVACIK 1986/5a EU528669 97.2 97.2 97.1 97.2 97.2 97.2 97.2 98.8 98.8 98.8 98.8 98.8 96.6 97.3 97.4 98.3 98.2
19 N. epilithica Kovacik 1998/7 HM018677 98.1 98.1 98.0 98.1 98.1 98.1 98.1 98.6 98.6 98.6 98.6 98.6 97.0 97.7 98.1 99.8 99.7 98.3
20 N. nodulosa UTEX 2910 EF122600 98.5 98.5 98.4 98.5 98.5 98.5 98.5 97.9 97.9 97.9 97.9 97.9 97.5 98.4 99.7 97.8 97.7 97.3 98.0
21 N. conica SEVA4-5-cl EU528667 97.4 97.4 97.3 97.4 97.4 97.4 97.4 97.0 97.0 97.0 97.0 97.0 96.5 97.2 97.4 98.4 98.3 96.6 98.2 97.3
22 N. ramsarensis KH-S S2.6 MF348318 98.8 98.8 98.7 98.8 98.8 98.8 98.8 97.8 97.8 97.8 97.8 97.8 97.4 98.9 98.8 97.3 97.2 97.0 97.5 99.1 96.8
23 N. radiophila TM S2B cl3 MF352005 98.2 98.2 98.1 98.2 98.2 98.2 98.2 97.3 97.3 97.3 97.3 97.3 96.9 98.3 98.0 97.0 96.9 96.5 97.0 97.9 96.5 98.2
24 N. svalbardensis 3320 MN567972 97.7 97.7 97.6 97.7 97.7 97.7 97.7 96.3 96.3 96.3 96.3 96.3 97.2 98.1 96.9 96.5 96.4 95.7 96.5 96.8 96.0 97.3 96.6
25 Nodosilinea sp. CXA007.4 KJ939094 97.8 97.8 97.7 97.8 97.8 97.8 97.8 98.7 98.7 98.7 98.7 98.7 96.9 97.8 97.8 99.5 99.4 98.4 99.3 97.7 97.9 97.6 97.1 96.4
26 Nodosilinea sp. IkpPStn44 KM438183 99.0 99.0 98.9 99.0 99.0 99.0 99.0 98.0 98.0 98.0 98.0 98.0 97.6 99.1 99.0 97.5 97.4 97.2 97.7 99.3 97.0 99.8 98.4 97.5 97.8
27 Nodosilinea sp. FI2-2HA2 HM018678 97.4 97.4 97.5 97.4 97.4 97.4 97.4 98.4 98.4 98.4 98.4 98.4 96.7 97.4 97.4 99.5 99.4 98.0 99.3 97.3 98.1 97.0 96.5 96.2 99.2 97.2

Sequences from Okinawan strains are in bold.

Table 3
Genetic similarity of all species of Nodosilinea calculated from sequences of 16S-23S ITS region
Strain 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22
1 N. coculeatus Ru1-19
2 N. coculeatus Ru1-11 100.0
3 N. coculeatus Ru1-18 100.0 100.0
4 N. coculeatus Ru1-4A 100.0 100.0 100.0
5 N. coculeatus Ru1-8 100.0 100.0 100.0 100.0
6 N. coculeatus Ru1-11-17 100.0 100.0 100.0 100.0 100.0
7 N. coculeatus Ru1-11-20 100.0 100.0 100.0 100.0 100.0 100.0
8 N. terrestrialis Ru4-3 92.7 92.7 92.7 92.7 92.7 92.7 92.7
9 N. terrestrialis Ru4-7 92.7 92.7 92.7 92.7 92.7 92.7 92.7 100.0
10 N. terrestrialis Ru4-17 92.7 92.7 92.7 92.7 92.7 92.7 92.7 100.0 100.0
11 N. terrestrialis Ru4-25 92.7 92.7 92.7 92.7 92.7 92.7 92.7 100.0 100.0 100.0
12 N. terrestrialis Ru4-26 92.7 92.7 92.7 92.7 92.7 92.7 92.7 100.0 100.0 100.0 100.0
13 Nodosilinea sp. ATA11-6K-CV21 KJ939096 91.4 91.4 91.4 91.4 91.4 91.4 91.4 95.6 95.6 95.6 95.6 95.6
14 N. bijugata KOVACIK1986/5a EU528669 90.7 90.7 90.7 90.7 90.7 90.7 90.7 96.5 96.5 96.5 96.5 96.5 94.1
15 Nodosilinea sp. ATA11-6B-CV9 KJ939097 89.2 89.2 89.2 89.2 89.2 89.2 89.2 92.7 92.7 92.7 92.7 92.7 93.0 91.9
16 Nodosilinea sp. IkpPStn44 KM438183 93.0 93.0 93.0 93.0 93.0 93.0 93.0 92.7 92.7 92.7 92.7 92.7 92.3 91.9 92.3
17 N. nodulosa UTEX B 2910 KF307598 91.9 91.9 91.9 91.9 91.9 91.9 91.9 91.6 91.6 91.6 91.6 91.6 92.5 90.1 89.9 91.6
18 N. epilithica Kovacik 1990/52 HM018679 91.4 91.4 91.4 91.4 91.4 91.4 91.4 92.1 92.1 92.1 92.1 92.1 90.5 92.1 89.6 92.7 92.1
19 Nodosilinea sp. WJT8-NPBG4 KJ939093 91.6 91.6 91.6 91.6 91.6 91.6 91.6 93.2 93.2 93.2 93.2 93.2 91.4 93.2 90.1 93.2 92.7 98.9
20 Nodosilinea sp. CMT-3FSIN-NPC22B KJ939095 91.9 91.9 91.9 91.9 91.9 91.9 91.9 91.6 91.6 91.6 91.6 91.6 91.4 92.1 88.3 91.2 94.3 94.5 95.2
21 Nodosilinea sp. FI2-2HA2 HM018678 91.9 91.9 91.9 91.9 91.9 91.9 91.9 93.4 93.4 93.4 93.4 93.4 91.2 93.8 90.3 92.1 92.3 95.8 96.9 96.7
22 Nodosilinea sp. CXA007.4 KJ939094 91.4 91.4 91.4 91.4 91.4 91.4 91.4 92.5 92.5 92.5 92.5 92.5 91.4 93.8 90.1 92.3 93.2 96.3 96.9 96.7 97.8
23 N. chupicuarensis KX859296 91.4 91.4 91.4 91.4 91.4 91.4 91.4 92.5 92.5 92.5 92.5 92.5 91.6 91.4 91.6 93.2 93.4 93.8 93.6 92.5 92.7 94.3

Sequences from Okinawan strains are in bold.

ITS, internal transcribed spacer.

Table 4
Identifiable domains in the 16S-23S rRNA ITS region from species of Nodosilinea
Strains Sequence length (bp) GenBank accession No. Reference

D1-D1′ tRNAIle V2 tRNAAla Box B V3
N. coculeatus Ru1-4A 61 73 - 72 40 27 LC731253 This study
N. coculeatus Ru1-8 61 73 - 72 40 27 LC731254 This study
N. coculeatus Ru1-11 61 73 - 72 40 27 LC731255 This study
N. coculeatus Ru1-11-17 61 73 - 72 40 26 LC731256 This study
N. coculeatus Ru1-11-20 61 73 - 72 40 26 LC731257 This study
N. coculeatus Ru1-18 61 73 - 72 40 27 LC731258 This study
N. coculeatus Ru1-19 61 73 - 72 40 27 LC731259 This study
N. terrestrialis Ru4-3 62 73 - 72 40 27 LC731260 This study
N. terrestrialis Ru4-7 62 73 - 72 40 27 LC731261 This study
N. terrestrialis Ru4-17 62 73 - 72 40 27 LC731262 This study
N. terrestrialis Ru4-25 62 73 - 72 40 27 LC731263 This study
N. terrestrialis Ru4-26 62 73 - 72 40 27 LC731264 This study
N. ramsarensis KH-S S2.6 61 73 - 72 47 27 - Heidari et al. (2018)
Nodosilinea sp. IkpPStn44 61 73 - 72 47 27 - Vázquez-Martínez et al. (2018)
N. epilithica Kovacik 1998/7 62 73 - 72 40 27 - Perkerson et al. (2011)
N. conica SEV4-5-c1 62 No data - No data 36 No data - Perkerson et al. (2011)
N. nodulosa UTEX2910 62 73 - 72 40 27 - Perkerson et al. (2011)
N. bijugata Kovacik 1986/5a 62 73 - 72 41 27 - Perkerson et al. (2011)
N. chupicuarensis PC471 62 73 - 72 40 27 - Vázquez-Martínez et al. (2018)
Nodosilinea sp. CXA007.4 62 73 - 72 40 27 - Vázquez-Martínez et al. (2018)
Nodosilinea sp. F1-2HA2 62 73 - 72 39 27 - Perkerson et al. (2011)
N. radiophila TM S2B 62 No data - No data 55 No data - Heidari et al. (2018)
N. hunanesis ZJJ01 62 73 - 72 47 27 - Cai et al. (2022)
N. svalbardensis 3220 62 73 - 72 47 27 - Davydov et al. (2020)

ITS, internal transcribed spacer.

Table 5
Morphological comparison among two new species of Nodosilinea from Okinawa and described species of Nodosilinea
Species Morphological characters Reference

Filaments Species Cell shape Cross-walls Cell width (μm) Cell length (μm) Apical cells
N. coculeatus sp. nov. Forming spiral shape under nitrogen-depleted conditions Generally thin, colorless Almost isodiametric, usually longer than wide Strongly constricted 1.2–1.5 1.4–2.7 Rounded This study
N. terrestrialis sp. nov. Forming nodules under nitrogen-depleted conditions Generally thin, colorless Almost isodiametric or sometimes longer than wide Constricted to distinctly constricted 1.2–1.7 1.2–2.2 Rounded This study
N. nodulosa Forming nodules Thin, colorless, occasionally becoming wide and diffluent Isodiametric, longer than wide Slightly constricted to strongly constricted 1.2–2.4 1.1–1.5 Rounded Perkerson et al. (2011)
N. signiensis Solitary, immotile, forming spiral Very thin, colorless Isodiametric, longer than wide / barrel shape Slightly constricted to strongly constricted 1.0 (1.5) 1.0–2.0 (2.3) Rounded Radzi et al. (2019)
N. epilithica Forming nodules in low light Thin, colorless, occasionally becoming wide and diffluent Barrel shaped, shorter to longer than wide Distinctly constricted 1.5–2.5 1.0–8.0 Rounded Perkerson et al. (2011)
N. bijugata Rarely forming nodules Often absent, thin, colorless Isodiametric, longer than wide Slightly constricted 1.5–1.7 1.5–6.2 Rounded Perkerson et al. (2011)
N. conica Rarely forming nodules Soft, thin, colorless Isodiametric, shorter than wide Slightly constricted 2.5–2.7 0.9–2.4 Rounded Perkerson et al. (2011)
N. chupicuarensis Multiseriate, motile, forming nodules Thin, clear Isodiametric Constricted 1.2 1.1–1.3 Dome-shaped Vázquez-Martínez et al. (2018)
N. ramsarensis Rarely forming nodules Thin, colorless Isodiametric, longer than wide Distinctly constricted 1.0–2.0 (0.8) 1.0–1.5 Not defined Heidari et al. (2018)
N. radiophila No. formation of nodules Thin, colorless Isodiametric, longer than wide Distinctly constricted 2.0–5.0 1.0–2.0 Rounded or elongated Heidari et al. (2018)
N. hunanesis Forming nodules Soft, layered, colorless, often becoming wide Cylindrical, longer than wide Strongly constricted 1.10–1.34 1.02–2.74 Dome-shaped or elongated Cai et al. (2022)
N. svalbardensis Forming nodules Soft, thin, colorless, sometimes widened, hyaline Shorter to longer than wide Strongly constricted 1.2–1.7 1.2–2.1 Rounded Davydov et al. (2020)

REFERENCES

Akagha, M. U., Pietrasiak, N., Bustos, D. F., Vondrášková, A., Lamb, S. C. & Johansen, J. R. 2023. Albertania and Egbenema gen. nov. from Nigeria and the United States, expanding biodiversity in the Oculatellaceae (Cyanobacteria). J. Phycol. 59:1217–1236. doi.org/10.1111/jpy.13389
crossref pmid
Berthold, D. E., Lefler, F. W. & Laughinghouse, D. H. 2022. Recognizing novel cyanobacterial diversity in marine benthic mats, with the description of Sirenicapillariaceae fam. nov., two new genera, Sirenicapillaria gen. nov. and Tigrinifilum gen. nov., and seven new species. Phycologia. 61:146–165. doi.org/10.1080/00318884.2021.2006589
crossref
Bohunická, M., Pietrasiak, N., Johansen, J. R., Gómez, E. B., Hauer, T., Gaysina, L. A. & et al 2015. Roholtiella, gen. nov. (Nostocales, Cyanobacteria): a tapering and branching cyanobacteria of the family Nostocaceae. Phytotaxa. 197:84–103. doi.org/10.11646/phytotaxa.197.2.2
crossref pdf
Boyer, S. L., Flechtner, V. R. & Johansen, J. R. 2001. Is the 16S-23S rRNA internal transcribed spacer region a good tool for use in molecular systematic and population genetics? A case study in cyanobacteria. Mol. Biol. Evol. 18:1057–1069. doi.org/10.1093/oxfordjournals.molbev.a003877
crossref pmid
Bravakos, P., Kotoulas, G., Skaraki, K., Pantazidou, A. & Economou-Amilli, A. 2016. A polyphasic taxonomic approach in isolated strains of cyanobacteria from thermal springs of Greece. Mol. Phylogenet. Evol. 98:147–160. doi.org/10.1016/j.ympev.2016.02.009
crossref pmid
Brito, Â, Ramos, V., Mota, R., Lima, S., Santos, A., Vieira, J. & et al 2017. Description of new genera and species of marine cyanobacteria from the Portuguese Atlantic coast. Mol. Phylogenet. Evol. 111:18–34. doi.org/10.1016/j.ympev.2017.03.006
crossref pmid
Brocke, H. J., Piltz, B., Herz, N., Abed, R. M. M., Palinska, K. A., John, U. & et al 2018. Nitrogen fixation and diversity of benthic cyanobacterial mats on coral reefs in Curaçao. Coral Reefs. 37:861–874. doi.org/10.1007/s00338-018-1713-y
crossref pdf
Cai, F., Li, S., Zhang, H., Yu, G. & Li, R. 2022. Nodosilinea hunanesis sp. nov. (Prochlorotrichaceae, Synechococcales) from a freshwater pond in China based on polyphasic approach. Diversity. 14:364 doi.org/10.3390/d14050364
crossref
Caires, T. A., Lyra, G. M., Hentschke, G. S., da Silva, A. M. S., de Araújo, V. L., Sant’Anna, C. L. & et al 2018. Polyphasic delimitation of a filamentous marine genus, Capillus gen. nov. (Cyanobacteria, Oscillatoriaceae) with the description of two Brazilian species. Algae. 33:291–304. doi.org/10.4490/algae.2018.33.11.25
crossref pdf
Casamatta, D. A., Gomez, S. R. & Johansen, J. R. 2006. Rexia erecta gen. et. sp. nov. and Capsosira lowei sp. nov, two newly described cyanobacterial taxa from the Great Smoky Mountains National Park (USA). Hydrobiology. 561:13–26. doi.org/10.1007/s10750-005-1602-6
crossref pdf
Charpy, L., Casareto, B. E., Langlade, M. J. & Suzuki, Y. 2012. Cyanobacteria in coral reef ecosystems: a review. J. Mar. Biol. 2012:259571 doi.org/10.1155/2012/259571
crossref pdf
Charpy, L., Palinska, K. A., Casareto, B., Langlade, M. J., Suzuki, Y., Abed, R. M. M. & et al 2010. Dinitrogen-fixing cyanobacteria in microbial mats of two shallow coral reef ecosystems. Microb. Ecol. 59:174–186. doi.org/10.1007/s00248-009-9576-y
crossref pmid pmc
Darriba, D., Taboada, G. L., Doallo, R. & Posada, D. 2012. jModelTest 2: more models, new heuristics and parallel computing. Nat. Methods. 9:772 doi.org/10.1038/nmeth.2109
crossref pmid pmc pdf
Davydov, D., Shalygin, S. & Vilnet, A. 2020. New cyanobacterium Nodosilinea svalbardensis sp. nov. (Prochlorotrichaceae, Synechococcales) isolated from alluvium in Mimer river valley of the Svalbard archipelago. Phytotaxa. 442:61–79. doi.org/10.11646/phytotaxa.442.2.2
crossref
Dvořák, P., Poulíčková, A., Hašler, P., Belli, M., Casamatta, D. A. & Papini, A. 2015. Species concepts and speciation factors in cyanobacteria, with connection to the problems of diversity and classification. Biodivers. Conserv. 24:739–757. doi.org/10.1007/s10531-015-0888-6
crossref pdf
Edgar, R. C. 2004. MUSCLE: multiple alignment with high accuracy and high throughput. Nucleic Acid Res. 32:1792–1797. doi.org/10.1093/nar/gkh340
crossref pmid pmc
Edler, D., Klein, J., Antonelli, A. & Silvestro, D. 2021. raxmlGUI 2.0: a graphical interface and toolkit for phylogenetic analyses using RAxML. Methods Ecol. Evol. 12:373–377. doi.org/10.1111/2041-210X.13512
crossref pdf
Gaydon, D. S., Probert, M. E., Buresh, R. J., Meinke, H. B. & Timsina, J. 2012. Modelling the role of algae in rich crop nutrition and soil organic carbon maintenance. Eur. J. Agron. 39:35–43. doi.org/10.7910/DVN/23785
crossref
González-Resendiz, L., Johansen, J. R., León-Tejera, H., Sánchez, L., Segal-Kischinevzky, C., Escobar-Sánchez, V. & et al 2019. A bridge too far in naming species: a total evidence approach does not support recognition of four species in Desertifilum (Cyanobacteria). J. Phycol. 55:898–911. doi.org/10.1111/jpy.12867
crossref pmid pdf
Gugger, M., Molica, R., Le Berre, B., Dufour, P., Bernard, C. & Humbert, J.-F. 2005. Genetic diversity of Cylindrospermopsis strains (Cyanobacteria) isolated from four continents. Appl. Environ. Microbiol. 71:1097–1100. doi.org/10.1128/AEM.71.2.1097-1100.2005
crossref pmid pmc pdf
Guindon, S. & Gascuel, O. 2003. A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst. Biol. 52:696–704, PMID: 10.1080/10635150390235520.
crossref pmid
Heidari, F., Zima, J., Riahi, H. & Hauer, T. 2018. New simple trichal cyanobacterial taxa isolated from radioactive thermal springs. Fottea. 18:137–149. doi.org/10.5507/fot.2017.024
crossref
Hoffmann, L. 1989. Algae of terrestrial habitats. Bot. Rev. 55:77–105.
crossref pdf
Hoffmann, L., Komárek, J. & Kaštovský, J. 2005. System of cyanoprokaryotes (cyanobacteria): state in 2004. Algol. Stud. 117:95–115. doi.org/10.1127/1864-1318/2005/0117-0095
crossref
Iteman, I., Rippka, R., de Marsac, N. T. & Herdman, M. 2000. Comparison of conserved structural and regulatory domains within divergent 16S rRNA-23S rRNA spacer sequences of cyanobacteria. Microbiology. 146:1275–1286. doi.org/10.1099/00221287-146-6-1275
crossref pmid
Johansen, J. R. & Casamatta, D. A. 2005. Recognizing cyanobacterial diversity through adoption of a new species paradigm. Algol. Stud. 116:71–93. doi.org/10.1127/1864-1318/2005/0117-0071
crossref
Johansen, J. R., Kovacik, L., Casamatta, D. A., Fučiková, K. & Kaštovský, J. 2011. Utility of 16S-23S ITS sequence and secondary structure for recognition of intrageneric and intergeneric limits within cyanobacterial taxa: Leptolyngbya corticola sp. nov. (Pseudanabaenaceae, Cyanobacteria). Nova Hedwigia. 92:283–302. doi.org/10.1127/0029-5035/2011/0092-0283
crossref
Jusko, B. M. & Johansen, J. R. 2023. Description of six new cyanobacterial species from soil biocrusts on San Nicolas Island, California, in three genera previously restricted to Brazil. J. Phycol. 60:133–151. doi.org/10.1111/jpy.13411
crossref pmid pdf
Kim, D.-H., Lee, N.-J., Kim, J.-H., Yang, E.-C. & Lee, O.-M. 2022. Three new Plectolyngbya species (Leptolyngbyaceae, Cyanobacteria) isolated from rocks and saltern of the Republic of Korea. Diversity. 14:1013 doi.org/10.3390/d14121013
crossref
Komárek, J. 2016. A polyphasic approach for the taxonomy of cyanobacteria: principles and applications. Eur. J. Phycol. 51:346–353. doi.org/10.1080/09670262.2016.1163738
crossref
Komárek, J. 2018. Several problems of the polyphasic approach in the modern cyanobacterial system. Hydrobiologia. 811:7–17. doi.org/10.1007/s10750-017-3379-9
crossref pdf
Komárek, J. & Anagnostidis, K. 2005. Süsswasserflora von Mitteleuropa. 19/2 Ed. Elsevier/Spektrum, Heidelberg, 759 pp.

Komárek, J., Kaštovský, J., Mareš, J. & Johansen, J. R. 2014. Taxonomic classification of cyanoprokaryotes (cyanobacterial genera) 2014, using a polyphasic approach. Preslia. 86:295–335.

Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. 2018. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 35:1547–1549. doi.org/10.1093/molbev/msy096
crossref pmid pmc
Lefler, F. W., Berthold, D. E. & Laughinghouse, H. D. 2021. The occurrence of Affixifilum gen. nov. and Neolyngbya (Oscillatoriaceae) in South Florida (USA), with the description of A. floridanum sp. nov. and N. biscaynensis sp. nov. J. Phycol. 57:92–110. doi.org/10.1111/jpy.13065
crossref pmid pdf
Lewin, R. A. 2006. Black algae. J. Appl. Phycol. 18:699–702. doi.org/10.1007/s10811-005-9018-2
crossref pdf
Mai, T., Johansen, J. R., Pietrasiak, N., Bohunická, M. & Martin, M. P. 2018. Revision of the Synechococcales (Cyanobacteria) through recognition of four families including Oculatellaceae fam. nov. and Trichocoleaceae fam. nov. and six new genera containing 14 species. Phytotaxa. 365:1–59. doi.org/10.11646/phytotaxa.365.1.1
crossref
Martins, M. D. & Branco, L. H. Z. 2016. Potamolinea gen. nov. (Oscillatoriales, Cyanobacteria): a phylogenetically and ecologically coherent cyanobacterial genus. Int. J. Syst. Evol. Microbiol. 66:3632–3641. doi.org/10.1099/ijsem.0.001243
crossref pmid
Martins, M. D., Machado-de-Lima, N. M. & Branco, L. H. Z. 2019. Polyphasic approach using multilocus analyses supports the establishment of the new aerophytic cyanobacterial genus Pycnacronema (Coleofasciculaceae, Oscillatoriales). J. Phycol. 55:146–159. doi.org/10.1111/jpy.12805
crossref pmid pdf
Miller, M. A., Pfeiffer, W. & Schwartz, T. 2010. Creating the CIPRES Science Gateway for inference of large phylogenetic trees. In: Proceedings of the Gateway Computing Environments Workshop (GCE); Curran Associates, Inc, New Orleans, LA, 1–8.
crossref
Mühlsteinová, R., Hauer, T., De Ley, P. & Pietrasiak, N. 2018. Seeking the true Oscillatoria: a quest for a reliable phylogenetic and taxonomic reference point. Preslia. 90:151–169. doi.org/10.23855/preslia.2018.151
crossref
Mühlsteinová, R., Johansen, J. R., Pietrasiak, N., Martin, M. P., Osorio-Santos, K. & Warren, S. D. 2014. Polyphasic characterization of Trichocoleus desertorum sp. nov. (Pseudanabaenales, Cyanobacteria) from desert soils and phylogenetic placement of the genus Trichocoleus . Phytotaxa. 163:241–261. doi.org/10.11646/phytotaxa.163.5.1
crossref pdf
Murakami, A., Mayashita, H., Iseki, M., Adachi, K. & Mimuro, M. 2004. Chlorophyll d in an epiphytic cyanobacterium of red algae. Science. 303:1633 doi.org/10.1126/science.1095459
crossref pmid
Neilan, B. A., Jacobs, D., Del Dot, T., Blackall, L. L., Hawkins, P. R., Cox, P. T. & et al 1997. rRNA sequences and evolutionary relationships among toxic and nontoxic cyanobacteria of the genus Mycrocystis . Int. J. Syst. Bacteriol. 47:693–697. doi.org/10.1099/00207713-47-3-693
crossref pmid
Nguyen, X. H., Sumimoto, S. & Suda, S. 2017. Unexpected high diversity of terrestrial cyanobacteria from the campus of the University of the Ryukyus, Okinawa, Japan. Microorganisms. 5:69 doi.org/10.3390/microorganisms5040069
crossref pmid pmc
Nübel, U., Garcia-Pichel, F. & Muyzer, G. 1997. PCR primers to amplify 16S rRNA genes from cyanobacteria. Appl. Environ. Microbiol. 63:3327–3332. doi.org/10.1128/aem.63.8.3327-3332.1997
crossref pmid pmc pdf
Nuryadi, H. & Suda, S. 2022. Revealing species diversity of Neolyngbya (Cyanobacteria, Oscillatoriales) from subtropical coastal regions of Okinawa, Japan, with description of Neolyngbya intertidalis sp. nov. and Neolyngbya latusa sp. nov. Phycol. Res. 70:69–80. doi.org/10.1111/pre.12482
crossref
Perkerson, R. B., Johansen, J. R., Kováčik, L., Brand, J., Kaštovský, J. & Casamatta, D. A. 2011. A unique Pseudanabaenalean (Cyanobacteria) genus Nodosilinea gen. nov. based on morphological and molecular data. J. Phycol. 47:1397–1412. doi.org/10.1111/j.1529-8817.2011.01077.x
crossref pmid
Pietrasiak, N., Osorio-Santos, K., Shalygin, S., Martin, M. P. & Johansen, J. R. 2019. When is a lineage a species?: a case study in Myxacorys gen. nov. (Synechococcales: Cyanobacteria) with the description of two new species from the Americas. J. Phycol. 55:976–996. doi.org/10.1111/jpy.12897
crossref pmid pdf
Radzi, R., Muangmai, N., Broady, P., Omar, W. M. W., Lavoue, S., Convey, P. & et al 2019. Nodosilinea signiensis sp. nov. (Leptolyngbyaceae, Synechococcales), a new terrestrial cyanobacterium isolated from mats collected on Signy Island, South Orkney Island, Antarctica. PLoS ONE. 14:e0224395 doi.org/10.1371/journal.pone.0224395
crossref pmid pmc
Řeháková, K., Johansen, J. R., Casamatta, D. A., Xuesong, L. & Vincent, J. 2007. Morphological and molecular characterization of selected desert soil cyanobacteria: three species new to science including Mojavia pulchra gen. et. sp. nov. Phycologia. 46:481–502. doi.org/10.2216/06-92.1
crossref
Ronquist, F. & Huelsenbeck, J. P. 2003. Mrbayes 3: Bayesian phylogenetic inference under mixed models. Bioinfromatics. 19:1572–1574. doi.org/10.1093/bioinformatics/btg180
crossref pmid pdf
Sciuto, K. & Moro, I. 2016. Detection of the new cosmopolitan genus Thermoleptolyngbya (Cyanobacteria, Leptolyngbyaceae) using the 16S rRNA gene and 16S-23S ITS region. Mol. Phylogenet. Evol. 105:15–35. doi.org/10.1016/j.ympev.2016.08.010
crossref pmid
Siegesmund, M. A., Johansen, J. R., Karsten, U. & Friedl, T. 2008. Coleofasciculus gen. nov. (Cyanobacteria): morphological and molecular criteria for revision of the genus Microcoleus Gomont. J. Phycol. 44:1572–1585. doi.org/10.1111/j.1529-8817.2008.00604.x
crossref pmid
Stackebrandt, E. & Ebers, J. 2006. Taxonomic parameters revisited tarnished gold standards. Microbiol. Today. 33:152–155.

Strunecký, O., Ivanova, A. P. & Mareš, J. 2023. An update classification of cyanobacterial orders and families based on phylogenomic and polyphasic analysis. J. Phycol. 59:12–51. doi.org/10.1111/jpy.13304
crossref pmid
Taton, A., Grubisic, S., Brambilla, E., De Wit, R. & Wilmotte, A. 2003. Cyanobacterial diversity in natural and artificial microbial mats of lake Fryxell (McMurdo Dry Valleys, Antarctica): a morphological and molecular approach. Appl. Environ. Microbiol. 69:5157–5169. doi.org/10.1128/AEM.69.9.5157-5169.2003
crossref pmid pmc pdf
Vaz, M. G. M. V., Genuário, D. B., Andreote, A. P. D., Malone, C. F. S., Sant’Anna, C. L., Barbiero, L. & et al 2015. Pantanalinema gen. nov. and Alkalinema gen. nov.: novel pseudanabaenacean genera (Cyanobacteria) isolated from saline-alkaline lakes. Int. J. Syst. Evol. Microbiol. 65:298–308. doi.org/10.1099/ijs.0.070110-0
crossref pmid
Vázquez-Martínez, J., Gutierrez-Villagomez, J. M., Fonseca-Gracía, C., Ramírez-Chávez, E., Mondragón-Sánchez, M. L., Partida-Martínez, L. & et al 2018. Nodosilinea chupicuarensis sp. nov. (Leptolyngbyaceae, Synechococcales) a subaerial cyanobacterium isolated from a stone monument in central Mexico. Phytotaxa. 334:167–182. doi.org/10.11646/phytotaxa.334.2.6
crossref pdf
Whitton, B. A. & Potts, M. 2000. The ecology of cyanobacteria: their diversity in time and space. Kluwer Academic Publishers, Dordrecht, 669 pp.

Yarza, P., Yilmaz, P., Pruesse, E., Glöckner, F. O., Ludwig, W., Schleifer, K.-H. & et al 2014. Uniting the classification of cultured and uncultured bacteria and archaea using 16S rRNA gene sequences. Nat. Rev. Microbiol. 12:635–645. doi.org/10.1038/nrmicro3330
crossref pmid pdf
Zammit, G., Billi, D. & Albertano, P. 2012. The subaerophytic cyanobacterium Oculatella subterranea (Oscillatoriales, Cyanophyceae) gen. et sp. nov.: a cytomorphological and molecular description. Eur. J. Phycol. 47:341–354. doi.org/10.1080/09670262.2012.717106
crossref
Zucker, M. 2003. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 31:3406–3415. doi.org/10.1093/nar/gkg595
crossref pmid pmc
TOOLS
PDF Links  PDF Links
PubReader  PubReader
ePub Link  ePub Link
Full text via DOI  Full text via DOI
Download Citation  Download Citation
  Print
Share:      
METRICS
0
Crossref
0
Scopus
652
View
52
Download
Related article
Editorial Office
[14348] A-1716, Gwangmyeong Trade Center, 72 Iljik-ro Gwangmyeong-si. Gyeonggi-do, Korea
Tel: +82-2-899-5980  Fax: +82-2-899-5922    E-mail: editalgae@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © The Korean Society of Phycology.                 Developed in M2PI