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      • KCI등재

        Improved Methodology for Identification of Cryptomonads: Combining Light Microscopy and PCR Amplification

        ( Shuang Xia ),( Ying Yin Cheng ),( Huan Zhu ),( Guo Xiang Liu ),( Zheng Yu Hu ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.3

        Cryptomonads are unicellular, biflagellate algae. Generally, cryptomonad cells cannot be preserved well because of their fragile nature, and an improved methodology should be developed to identify cryptomonads from natural habitats. In this study, we tried using several cytological fixatives, including glutaraldehyde, formaldehyde, and their combinations to preserve field samples collected from various waters, and the currently used fixative, Lugol`s solution was tested for comparison. Results showed that among the fixatives tested, glutaraldehyde preserved the samples best, and the optimal concentration of glutaraldehyde was 2%. The cell morphology was well preserved by glutaraldehyde. Cells kept their original color, volume, and shape, and important taxonomic features such as furrow/gullet complex, ejectosomes, as well as flagella could be observed clearly, whereas these organelles frequently disappeared in Lugol`s solution preserved samples. The osmotic adjustments and buffers tested could not preserve cell density significantly higher. Statistical calculation showed the cell density in the samples preserved by 2% glutaraldehyde remained stable after 43 days of the fixation procedure. In addition, DNA was extracted from glutaraldehyde preserved samples by grinding with liquid nitrogen and the 18S rDNA sequence was amplified by PCR. The sequence was virtually identical to the reference sequence, and phylogenetic analyses showed very close relationship between it and sequences from the same organism. To sum up, the present study demonstrated that 2% unbuffered glutaraldehyde, without osmotic adjustments, can preserve cryptomonads cells for identification, in terms of both light microscopy and phylogenetic analyses based on DNA sequences.

      • KCI등재

        온천천 하류 적조 원인생물의 동정 및 발생 특성

        김미희,지희성,조정구,조순자 한국물환경학회 2018 한국물환경학회지 Vol.34 No.3

        This study was performed in order to identify the red tide-causing organism and to understand the characteristics of the water quality during the winter of 2015 and 2016 in the Oncheon stream, a tidal river in Busan, where red tide often occurs in the wintertime. Two sites were selected on the stream and the surface water was sampled a total of 28 times during the experimental period. Twelve water quality characteristics, including water temperature, pH, DO, COD, total-N (T-N), total-P (T-P), and salinity were analyzed in order to test water quality. The cell numbers of cryptomonads were counted directly by microscopic observation. The nucleotide sequences of the partial 28S rRNA gene and psbA gene from metagenomic DNA, derived from each sampling site, were analyzed. According to the results, the alga most responsible for the bloom was identified as Teleaulax OC1 sp., which belongs to the cryptomonads. Three items of chl-a, pH, and DO were positively correlated with the cell numbers of the cryptomonads counted at the upper stream of the tidal area (St 1) while eight items of chl-a, TOC, BOD, total-N, COD, SS, pH, and DO were positively correlated with the cells located at the junction between the stream and Su-young river (St 2) in the order.

      • KCI등재

        Ultrastructure of the flagellar apparatus in Rhinomonas reticulata var. atrorosea (Cryptophyceae, Cryptophyta)

        남승원,고동희,손미선,신웅기 한국조류학회I 2013 ALGAE Vol.28 No.4

        Rhinomonas reticulata var. atrorosea G. Novarino is a photosynthetic marine flagellate that is known to have typical characteristics of cryptomonads. We examined the flagellar apparatus of R. reticulata var. atrorosea by transmission electron microscopy. The major components of the flagellar apparatus of R. reticulata var. atrorosea consisted of four types of microtubular roots (1r, 2r, 3r, and mr), a non-keeled rhizostyle (Rhs), mitochondrion-associated lamella (ML), two connections between basal bodies, a striated fibrous root (SR) and a striated fiber-associated microtubular root (SRm). Four types of microtubular roots originated near the ventral basal body and extended toward the left side of the basal bodies. The non-keeled Rhs originated at the Rhs-associated striated fiber, which was located between two basal bodies and extended into the middle of the cell. The ML was a plate-like fibrous structure associated with mitochondria and originating from a Rhs-associated fiber. It split into two parts and extended toward the dorsal-posterior of the cell to a mitochondrion. The SR and SRm extended parallel to the anterior lobe of the cell. The overall configuration of the flagellar apparatus in R. reticulata var. atrorosea was similar to the previously reported descriptions of those of Cryptomonas paramecium, C. pyrenoidifera, C. ovata, Hanusia phi, Guillardia theta, and Proteomonas sulcata. However, the flagellar apparatus system of R. reticulata var. atrorosea was more complex than those of other cryptomonad species due to the presence of an additional microtubular root and other distinctive features, such as a rhizostyle-associated striated fiber and large ML.

      • SCIESCOPUSKCI등재

        Ultrastructure of the flagellar apparatus in Rhinomonas reticulata var. atrorosea (Cryptophyceae, Cryptophyta)

        Nam, Seung Won,Go, Donghee,Son, Misun,Shin, Woongghi The Korean Society of Phycology 2013 ALGAE Vol.28 No.4

        Rhinomonas reticulata var. atrorosea G. Novarino is a photosynthetic marine flagellate that is known to have typical characteristics of cryptomonads. We examined the flagellar apparatus of R. reticulata var. atrorosea by transmission electron microscopy. The major components of the flagellar apparatus of R. reticulata var. atrorosea consisted of four types of microtubular roots (1r, 2r, 3r, and mr), a non-keeled rhizostyle (Rhs), mitochondrion-associated lamella (ML), two connections between basal bodies, a striated fibrous root (SR) and a striated fiber-associated microtubular root (SRm). Four types of microtubular roots originated near the ventral basal body and extended toward the left side of the basal bodies. The non-keeled Rhs originated at the Rhs-associated striated fiber, which was located between two basal bodies and extended into the middle of the cell. The ML was a plate-like fibrous structure associated with mitochondria and originating from a Rhs-associated fiber. It split into two parts and extended toward the dorsal-posterior of the cell to a mitochondrion. The SR and SRm extended parallel to the anterior lobe of the cell. The overall configuration of the flagellar apparatus in R. reticulata var. atrorosea was similar to the previously reported descriptions of those of Cryptomonas paramecium, C. pyrenoidifera, C. ovata, Hanusia phi, Guillardia theta, and Proteomonas sulcata. However, the flagellar apparatus system of R. reticulata var. atrorosea was more complex than those of other cryptomonad species due to the presence of an additional microtubular root and other distinctive features, such as a rhizostyle-associated striated fiber and large ML.

      • KCI등재

        New records of the genus Chroomonas and two Chroomonas species (Cryptomonadales, Cryptophyceae) from Korean freshwater

        Hyeon Ju Nam,Miran Kim,Seok Won Jang,Bok Yeon Jo,Eunyoung Moon,Seung Won Nam 한국환경생물학회 2021 환경생물 : 환경생물학회지 Vol.39 No.4

        The genus Chroomonas is a group of blue-green colored cryptomonads. This study describes two freshwater Chroomonas species for the first time in South Korea: Chroomonas nordstedtii Hansgirg and Chroomonas coerulea (Geitler) Skuja. We examined the morphology and ultrastructure of these species by light microscopy, scanning electron microscopy, and transmission electron microscopy. These two Chroomonas species were blue-green colored and ovate to oval-shaped. Chroomonas nordstedtii was characterized by two Maupas ovals with hexagonal periplast plates, whereas C. coerulea was characterized by one eyespot with rectangular periplast plates. A molecular phylogeny with data from nuclear SSU rRNA and chloroplast rbcL genes revealed that Korean C. nordstedtii formed a distinct clade with NIES-708, NIES-1004 from Japan, and UTEX 2779 from Colorado, USA, while C. coerulea formed a clade with ACOI 1366 from Portugal.

      • KCI등재

        Ultrastructure of the flagellar apparatus in Rhodomonas salina (Cryptophyceae, Cryptophyta)

        Nam, Seung Won,Jo, Bok Yeon,Shin, Woongghi Korean Society of Environmental Biology 2020 환경생물 : 환경생물학회지 Vol.38 No.2

        Rhodomonas salina is a phototrophic marine flagellate. We examined the ultrastructure of R. salina with particular attention to the flagellar apparatus by transmission electron microscopy and compared it with that of other cryptomonads reported previously. The major components of the flagellar apparatus in R. salina were a keeled rhizostyle (Rhs), a striated fibrous root(SR), a SR-associated microtubular root (SRm), a mitochondrion-associated lamella (ML), and three types of microtubular roots (9r, 4r, and 2r). The keeled Rhs originated near the proximal end of the dorsal basal body, passed near the nucleus and dissociated at the posterior end of the cell. The SR and SRm originated between two basal bodies and laterally extended to the right side of the cell. The ML originated between two basal bodies and extended to the left side of the cell. The 9r originated between the ventral basal body and the Rhs and extended toward the anterior dorsal lobe of the cell. The 4r originated near the 9r and extended toward the dorsal lobe with the 2r, which originated between two basal bodies. Here, the flagellar apparatus in R. salina is described, and the ultrastructure of the flagellar apparatus is compared among cryptomonad species.

      • KCI등재
      • SCIESCOPUSKCI등재

        Intraspecific variations in macronutrient, amino acid, and fatty acid composition of mass-cultured Teleaulax amphioxeia (Cryptophyceae) strains

        Lee, Bae Ik,Kim, Shin Kwon,Kim, Jong Hyeok,Kim, Hyung Seop,Kim, Jong Im,Shin, Woongghi,Rho, Jung-Rae,Yih, Wonho The Korean Society of Phycology 2019 ALGAE Vol.34 No.2

        To compare the nutritional quality of TPG (Teleaulax / Plagioselmis / Geminigera) clade species of cryptomonads with that of RHO (Rhodomonas / Rhinomonas / Storeatula) clade species 6 Teleaulax amphioxeia (TA) and 1 Rhinomonas sp. strains were mass-cultured in newly designed 500-L photobioreactors to the end of exponential growth phase. Intraspecific variations (IVs) in terms of one standard deviation among the 6 TA strains in the compositions of the three macronutrients were 41.5 (protein), 89.8 (lipid), and 15.6% (carbohydrate) of the mean. When harvested from stationary growth phase mean compositions of essential amino acids (EAAs, 47.3%) and non-EAAs (52.7%) of the 2 TA strains, CR-MAL07 and CR-MAL08-2, were similar to those of a Chroomonas strain. The IVs between the 2 TA strains in the composition of EAAs (10.3 and 2.4) and non-EAAs (8.5 and 2.1% of the mean) were rather smaller than those of saturated fatty acids (30.3 and 26.1) and unsaturated fatty acids (UFAs, 12.0 and 12.5% of the mean) in f/2-Si and urea-based compound fertilizer (UCF) culture media, respectively. Mean compositions of eicosapentaenoic acid (EPA, 17.9%) and docosahexaenoic acid (DHA, 12.7%) of total fatty acids of the 2 TA strains were higher than those that of a Chroomonas strain. EPA and DHA compositions exhibited similar level of IVs between the 2 TA strains in f/2-Si (14.6 and 11.0) and UCF media (12.6 and 13.5% of the mean). Thus, the nutritional quality in terms of amino acids, UFAs, EPA, and DHA in a TPG clade species, T. amphioxeia was comparable to those of RHO clade species with notable IVs. Practically, biotechnological targets for TPG clade cryptomonad strains might be subspecies or clone level.

      • KCI등재

        Ultrastructure of the flagellar apparatus in cryptomorphic Cryptomonas curvata (Cryptophyceae) with an emphasis on taxonomic and phylogenetic implications

        남승원,신웅기 한국조류학회I 2016 ALGAE Vol.31 No.2

        Cryptomonas curvata Ehrenberg is a photosynthetic freshwater flagellate and the type species of the genus Cryptomonas. We examined the flagellar apparatus of cryptomorphic C. curvata by transmission electron microscopy. The major components of the flagellar apparatus are the non-keeled rhizostyle (Rhs), striated fibrous root (SR), striated fiber-associated microtubular root (SRm), mitochondrion-associated lamella (ML), and two types of microtubular roots (3r and 2r). The non-keeled Rhs originate at the ventral basal body and consist of two types of microtubule bands extending together into the middle of the cell. The SR and SRm extend parallel to the left side of the cell. The ML originates from the ventral basal body and is a plate-like fibrous structure associated with mitochondria. The 3r extends from the dorsal basal body toward the dorsal anterior of the cell. The 2r originates between the two basal bodies and extends shortly to the left of the cell. The overall configuration of the flagellar apparatus is most similar to that previously reported for C. pyrenoidifera. These results demonstrate that the features of the flagellar apparatus are useful for distinguishing closely related species and inferring phylogenetic relationships among taxa.

      • KCI등재

        Intraspecific variations in macronutrient, amino acid, and fatty acid composition of mass-cultured Teleaulax amphioxeia (Cryptophyceae) strains

        Bae Ik Lee,Shin Kwon Kim,Jong Hyeok Kim,Hyung Seop Kim,Jong Im Kim,Woongghi Shin,Jung-Rae Rho,Wonho Yih 한국조류학회I 2019 ALGAE Vol.34 No.2

        To compare the nutritional quality of TPG (Teleaulax / Plagioselmis / Geminigera) clade species of cryptomonadswith that of RHO (Rhodomonas / Rhinomonas / Storeatula) clade species 6 Teleaulax amphioxeia (TA) and 1 Rhinomonassp. strains were mass-cultured in newly designed 500-L photobioreactors to the end of exponential growth phase. Intraspecific variations (IVs) in terms of one standard deviation among the 6 TA strains in the compositions of the threemacronutrients were 41.5 (protein), 89.8 (lipid), and 15.6% (carbohydrate) of the mean. When harvested from stationarygrowth phase mean compositions of essential amino acids (EAAs, 47.3%) and non-EAAs (52.7%) of the 2 TA strains,CR-MAL07 and CR-MAL08-2, were similar to those of a Chroomonas strain. The IVs between the 2 TA strains in thecomposition of EAAs (10.3 and 2.4) and non-EAAs (8.5 and 2.1% of the mean) were rather smaller than those of saturatedfatty acids (30.3 and 26.1) and unsaturated fatty acids (UFAs, 12.0 and 12.5% of the mean) in f/2-Si and urea-basedcompound fertilizer (UCF) culture media, respectively. Mean compositions of eicosapentaenoic acid (EPA, 17.9%) anddocosahexaenoic acid (DHA, 12.7%) of total fatty acids of the 2 TA strains were higher than those that of a Chroomonasstrain. EPA and DHA compositions exhibited similar level of IVs between the 2 TA strains in f/2-Si (14.6 and 11.0) andUCF media (12.6 and 13.5% of the mean). Thus, the nutritional quality in terms of amino acids, UFAs, EPA, and DHA ina TPG clade species, T. amphioxeia was comparable to those of RHO clade species with notable IVs. Practically, biotechnologicaltargets for TPG clade cryptomonad strains might be subspecies or clone level.

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