• Environmental Engineering Research
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• v.19 no.4
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• pp.317-329
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• 2014

The diversity of cyanobacteria in Sri Lanka was studied in different water reservoirs, paddy fields, brackish water and tsunami affected areas using light microcopy, 16S rRNA sequences, followed by phylogenetic analysis. Based on light microscopy, 24 genera were identified from environmental samples belonging to the orders Chroococcales, Oscillatoriales, Pleurocapsales and Nostocales. In cultures, 33 genera were identified from all five cyanobacterial orders, including Stigonematales. Based on 16S rRNA gene sequences and their morphology, two isolates were identified up to species level, 72 to genus level, one isolate up to family and 11 up to order level. Twelve isolates couldn't be assigned to any taxonomic level. The results of 16S rRNA gene sequences along with the phylogenetic analysis indicated that some cyanobacterial isolates could be accommodated to genus or order level. The 16S rRNA sequence analysis data in this study confirmed that order Nostocales and order Pleurocapsales cyanobacteria are monophyletic while orders Chroococcales, Oscillatoriales and Stigonematales cyanobacteria are polyphyletic. Polyphasic approach including the combination of light microscopy, cultures and the analysis of 16S rRNA gene sequences provide a promising approach to ascertain the diversity of cyanobacteria in different habitats.

• Korean Journal of Microbiology
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• v.47 no.3
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• pp.268-274
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• 2011

Anabaena (Cyanobacteria, Nostocales) are important for water quality controls, because they are often responsible for freshwater green tides; moreover, some species are reported to produce hepatotoxin. In this study, we sequenced RNA polymerase beta subunit (rpoB) gene of Anabaena, and evaluated their sequences for the potential use of a molecular taxonomic marker in this taxon. Anabaena rpoB showed low DNA similarity and high genetic divergences when compared those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.01). Parsimony analyses showed the rpoB gene evolves 4.8-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each Anabaena strain more clearly compared with a 16S rRNA tree. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the species discrimination of Anabaena.

• Journal of Korean Society on Water Environment
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• v.32 no.1
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• pp.52-59
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• 2016

This study investigated the occurrences and succession patterns of harmful cyanobacteria, as well as environmental factors, during a 3-year period (September 2012 to August 2015) in the upper region of the Nakdong River around Sangju weir. A total of 27 cyanobacterial taxa were observed in this study, and classified into 26 species and 1 variety belonging to 11 genera, 5 families, and 3 orders. Cell density ranged from 24 to 42,001 cells/ml, with a geometric mean of 33 cells/ml, during the survey period. The dominant orders differed depending on the survey year; order Oscillatoriales in 2013, Chroococcales in 2014 and Nostocales in 2015. An Aphanizomenon bloom occurred in June 2015 at which time the highest cell density of 36,873 cells/ml was detected in the upper region of the Nakdong River, where as the Aphanizomenon spp. cell density (190-1,704 cells/ml) had been low prior to that time. An Aphanizomenon bloom also occurred at around the same time downstream in the Young River, a major inflow branch of the Nakdong River. The Aphanizomenon cell density along the Nakdong River increased markedly after joining of the YoungRiver, indicating that the Aphanizomenon bloom in the YoungRiver caused a bloom in the Nakdong River. Meteorological and environmental parameters, such as very low precipitation, higher water temperature, pH, and TP concentration, and lower TN/TP ratio, in May and June of 2015 than in 2013 and 2014 exerted marked effects on the Aphanizomenon bloom in June 2015 in the Young River.

• Journal of Korean Society on Water Environment
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• v.34 no.5
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• pp.503-513
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• 2018

Freshwater cyanobacterial genus Dolichospermum is one of the most commonly spotted types of phytoplankton, whereas a limited number of studies on morphology of Dolichospermum spp. have been performed in South Korea. The purpose of this study is to investigate the succession pattern of cyanobacteria after weir construction, as well as morphological characteristics of Dolichospermum spp. from natural samples collected in the weir regions of Nakdong River. A total of 31 cyanobacterial taxa observed in this study were classified as belonging to 15 genera, 5 families, and 3 orders. Among them, morphological characteristics in the four species were classified into genus Dolichospermum, for most of the planktic former members of the genus Anabaena, were observed through light microscopy. Water bloom frequently occurred in the middle region of Nakdong River, the maximum number of cyanobacterial species appeared in the lower region of Nakdong River. The appearance of order Chrooccocales was only observed during summer when population density of Microcystis aeruginosa reached an annual peak. In contrast, filamentous cyanobacteria was observed throughout the whole year, even if when water temperature was lower than $5^{\circ}C$. It implied that the low-temperature-adapted filamentous cyanobacteria can grow in a range of water temperatures. Coil diameter of D. crassum from natural samples was $75{\sim}140{\mu}m$ ($ave.=91.3{\mu}m$; n = 94), slightly larger than those reported by previous studies. Dolichospemum smithii ($Kom{\grave{a}}rek$) Wacklin et al. 2009, was described for the first time in Nakdong River.

• Journal of Korean Society on Water Environment
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• v.34 no.6
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• pp.661-668
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• 2018

Geosmin is volatile metabolites produced by a range of filamentous cyanobacteria which causes taste and odor problems in drinking water. Molecular ecological methods which target biosynthetic genes (geoA) are widely adopted to detect geosmin-producing cyanobacteria. The aim of this study was to investigate the potential production capability of 8 strains isolated from the Nakdong River. Ultimately, a suggestion for a genetical monitoring tool for the identification of geosmin producers in domestic waters was to be made. Geosmin was detected using solid phase microextraction gas chromatography mass spectrometry (SPME GC-MS) in two strains of Dolichospermum plactonicum (DGUC006, DGUC012) that were cultured for 28 day. The highest concentrations during the experiment period was $17,535ngL^{-1}$ and $14,311ngL^{-1}$ respectively. Additionally, geoA genes were amplified using two primers (geo78F/971R and geo78F/982R) from strains shown to produce geosmin, while amplification products were not detected in any of non-producing strains. PCR product (766 bp) was slightly shorter than the expected size for geosmin producers. According to the BLAST analysis, amplified genes were at nucleotide level with Anabaena ucrainica (HQ404996, HQ404997), Dolichospermum planctonicum (KM13400) and Dolichospermum ucrainicum (MF996872) between 99 ~ 100 %. Both strains were thus confirmed as potential geosmin-producing species. We concluded that the molecular method of analysis was a useful tool for monitoring potential cyanobacterial producers of geosmin.

• ALGAE
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• v.35 no.1
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• pp.1-15
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• 2020

Two strains isolated from a subtropical region in China, were morphologically identified as a Nostoc-like species, but its taxonomic identity was unknown. In this study, these two strains were taxonomically and phylogenetically characterized based on polyphasic approach combining morphological and genetic characteristics. Though both were virtually indistinguishable from Nostoc in field and cultured material, these two strains were phylogenetically distinct from Nostoc based on 16S rRNA phylogeny. The 16S-23S internal transcribed spacer rRNA secondary structure of these strains showed the unique pattern of D1-D1', Box-B, and V3 helix, which distinguished them from other Nostoc-like heterocytous genera. A unique cluster separated from Nostoc sensu stricto supports the establishment of Violetonostoc gen. nov. with the type species as Violetonostoc minutum sp. nov.

• Journal of Ecology and Environment
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• v.40 no.3
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• pp.144-152
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• 2016

Background: The purpose of this study is to describe the morphological characteristics of the Aphanizomenon spp. and related species from the natural samples collected in the Nakdong River of South Korea. Results: Morphological characteristics in the four species classified into the genera Aphanizomenon Morren ex Bornet et Flahault 1888 and Cuspidothrix Rajaniemi et al. 2005 were observed by light microscopy. The following four taxa were identified: Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault, Aphanizomenon klebahnii Elenkin ex Pechar, Aphanizomenon skujae $Kom{\acute{a}}rkov{\acute{a}}-Legnerov{\acute{a}}$ et Cronberg, and Cuspidothrix issatschenkoi ($Usa{\check{c}}ev$) Rajaniemi et al. Aph. flos-aquae and Aph. klebahnii always formed in fascicles; the others only occurred in solitary. Aph. flos-aquae was similar to Aph. klebahnii, whereas these species differed from each other by the size and shape of fascicles, which was macroscopic in Aph. flos-aquae and microscopic in the Aph. klebahnii. One of their characteristics was that trichomes are easily disintegrating during microscopic examination. C. issatschenkoi could be clearly distinguished from other species by hair-shaped terminal cell. Its terminal cell was almost hyaline and markedly pointed. Young populations of the species without heterocytes run a risk of a misidentification. Aph. skujae was characterized by akinete. Morphological variability of akinetes from natural samples collected in the Nakdong River was rather smaller than those reported by previous study. Conclusions: C. issatschenkoi are described for the first time in the Nakdong River. In addition, Aph. klebahnii and Aph. skujae are new to South Korea.

• ALGAE
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• v.33 no.2
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• pp.143-156
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• 2018

Dolichospermum is a filamentous and heterocytous cyanobacterium that is one of the commonly occurring phytoplanktons in the Han River of Korea. Morphological observations led to the identification of D. planctonicum-like filaments in seasonal water samples. In the present study, we successfully isolated these filaments using culture methods, and examined its morphology using light and scanning electron microscopy. The morphology of the D. planctonicumlike species differed from that of typical D. planctonicum; it had thin cylindrical-shaped akinetes, which were narrower towards the ends than at the center. This morphology is firstly described in the genus Dolichospermum. In addition, the akinetes in the filament developed solitarily and were distant from the heterocytes. Phylogenetic analysis of the 16S rRNA sequences showed that our Dolichospermum clustered with D. planctonicum and D. circinale, which have coiled trichome. However, phylogenetic analysis of the gene encoding rivulose-1,5-bisphosphate carboxylase (rbcLX) clearly separated our species from other Dolichospermum, forming a unique clade. Additionally, structures of D. planctonicum and D. hangangense strains were different type in Box-B and V3 region. These results demonstrated that the new Dolichospermum species was unique in morphology and molecular traits. Therefore, we propose this to be a new species belonging to genus Dolichospermum with the name Dolichospermum hangangense sp. nov.

• ALGAE
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• v.37 no.1
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• pp.1-14
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• 2022

Cyanobacteria are distributed worldwide, and many new cyanobacterial species are discovered in tropical region. The Nostoc-like genus Amazonocrinis has been separated from the genus Nostoc based on polyphasic methods. However, species diversity within this genus remains poorly understood systematically because only one species (Amazonocrinis nigriterrae) has been described. In this study, two novel strains (NUACC02 and NUACC03) were isolated from moist rice field soil in Thailand. These two strains were characterized using a polyphasic approach, based on morphology, 16S rRNA phylogenetic analysis, internal transcribed spacer secondary structure and ecology. Phylogenetic analyses based on 16S rRNA gene sequences confirmed that the two novel strains formed a monophyletic clade related to the genus Amazonocrinis and were distant from the type species A. nigriterrae. The 16S rRNA gene sequence similarity (<98.1%) between novel strains and all other closely related taxa including the Amazonocrinis members exceeded the cutoff for species delimitation in bacteriology, reinforcing the presence of a new Amazonocrinis species. Furthermore, the novel strains possessed unique phenotypic characteristics such as the presence of the sheath, necridia-like cells, larger cell dimension and akinete cell arrangement in long-chains and the singularity of D1-D1', Box-B, V2, and V3 secondary structures that distinguished them from other Amazonocrinis members. Considering all the results, we described our two strains as Amazonocrinis thailandica sp. nov. in accordance with the International Code of Nomenclature for Algae, Fungi and Plants.

• Korean Journal of Ecology and Environment
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• v.50 no.1
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• pp.137-147
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• 2017

The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.