• Title/Summary/Keyword: polyphasic study

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Six newly recorded species of epilithic cyanobacteria isolated in Korea

  • Hye-Ryeung Wang;Ji-Ho Song;Nam-Ju Lee;Do-Hyun Kim;So-Won Kim;Ok-Min Lee
    • Journal of Species Research
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    • v.13 no.1
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    • pp.10-31
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    • 2024
  • In this study, 11 strains of epilithic cyanobacteria belonging to six unrecorded species in Korea were isolated from gravel submerged in freshwater of seven collection sites in Korea. The morphological characteristics of the six species isolated in this study were consistent with the type strain of each species, and the similarity of the 16S rRNA gene sequences with the type strain of each species were 98.8-100%. In the phylogenetic tree using the 16S rRNA gene sequences, the 11 strains of these six species formed the same cluster as the strains of each species. The habitat of each previously reported species is mainly the soil surface, but all Korean strains appeared from the gravel submerged in freshwater. As a result of the morphological, ecological, and molecular analyses, these six species of cyanobacteria were identified as Geminocystis papuanica, Allocoleopsis franciscana, Ancylothrix terrestris, Klisinema persicum, Scytolyngbya timoleontis, and Shackletoniella antarctica, which were added as newly recorded species in Korea.

Identification and Distribution of predominant tactic Acid Bacteria in Kimchi, a Korean Traditional Fermented Food

  • Kim, Tae-Woon;Lee, Ji-Yeon;Jung, Sang-Hoon;Kim, Young-Mok;Jo, Jae-Sun;Chung, Dae-Kyun;Lee, Hyong-Joo;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.635-642
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    • 2002
  • To effectively investigate the identification and distribution of the lactic acid bacteria in Kimchi, polyphasic methods, including a PCR, SDS-PAGE of the whole-cell proteins, and 16S rRNA gene sequence analysis, were used. In various types of Kimchi fermented at 20$\^{C}$, the isolate KHU-31 was found to be the predominant lactic acid bacteria. This isolate was identified as Lactobacillus sake KHU-31, based on SDS-PAGE of the whole-cell proteins and a 165 rRNA gene sequence analysis, which provided accurate and specific results. Accordingly, the approach used in the current study demonstrated that Lactobacillus sake KHU-31, together with Leuconostoc mesenteroides, were the most predominant lactic acid bacteria in all types of Kimchi in the middle stage of fermentation at 20$\^{C}$.

Genetic Diversity and Molecular Phylogeny of Cyanobacteria from Sri Lanka Based on 16S rRNA Gene

  • Wanigatunge, R.P.;Magana-Arachchi, D.N.;Chandrasekharan, N.V.;Kulasooriya, S.A.
    • 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.

Thalassobius aestuarii sp. nov., Isolated from Tidal Flat Sediment

  • Yi Ha-Na;Chun Jong-Sik
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.171-176
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    • 2006
  • A strictly aerobic, non-motile, ovoid-shaped Alphaproteobacteria, designated strain $JC2049^T$ was isolated from a tidal flat sediment sample. The results of 16S rRNA gene sequence analysis indicated that this isolate belonged to the genus Thalassobius, with a sequence similarity of 96.9-97.3% to other valid Thalassobius spp. The cells required 1-7% NaCl for growth (optimum 2%) and accumulated $poly-\beta-hydroxybutyrate$. Nitrite was reduced to nitrogen, but nitrate was not reduced to nitrite. No genetic potential for aerobic anoxygenic photosynthesis was detected. The primary isoprenoid quinone (Ubiquinone-10), predominant cellular fatty acids $(C_{18:1}{\omega}7c,\;11\;methyl\;C_{18:1}\omega7c\;and\;C_{16:0})$ and DNA G+C content (61 mol %) were all consistent with the assignment of this isolate to the genus Thalassobius. Several phenotypic characteristics clearly distinguished our isolate from other Thalassobius species. The degree of genomic relatedness between strain $JC2049^T$ and other Thalassobius species was in a range of 20-43 %. The polyphasic data presented in this study indicates that our isolate should be classified as a novel species within the genus Thalassobius. The name Thalassobius aestuarii sp. novo is therefore proposed for this isolate; the type strain is $JC2049^T(=IMSNU\;14011^T=KCTC\;12049^T=DSM\;15283^T)$.

Polyphasic Microbial Analysis of Traditional Korean Jeung-Pyun Sourdough Fermented with Makgeolli

  • Lim, Sae Bom;Tingirikari, Jagan Mohan Rao;Kwon, Ye Won;Li, Ling;Kim, Grace E.;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.226-233
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    • 2017
  • Jeung-pyun, a fermented rice cake, is prepared by fermenting rice sourdough using makgeolli, a traditional Korean rice wine, in the presence of yeast and lactic acid bacteria (LAB). The goal of this study was to conduct biochemical and microbial analyses of five different rice sourdoughs, each fermented with a different commercial makgeolli, using culture-dependent and culture-independent approaches. All sourdough samples fermented with different makgeolli for 6.5 h showed different profiles in pH, total titratable acidity, organic acid concentration, and microbial growth. LAB belonging to different genera were identified based on colony morphology on modified MRS and sourdough bacteria agar medium. PCR-denaturing gradient gel electrophoresis analyses of the five sourdoughs showed different bands corresponding to LAB and yeast. 16S/26S rRNA gene sequence analyses of the samples confirmed that the predominant LAB in the five fermented rice doughs was Lactobacillus plantarum, Lb. pentosus, and Lb. brevis. Various other Lactobacillus spp. and Saccharomyces cerevisiae were common in all five fermented samples. This study provides comprehensive and comparative information on the microflora involved in fermentation of rice sourdough and signifies the need to develop effective starters to enrich the quality of jeung-pyun.

Isolation, Characterization and Whole-Genome Analysis of Paenibacillus andongensis sp.nov. from Korean Soil

  • Yong Guan;Zhun Li;Yoon-Ho Kang;Mi-Kyung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.753-759
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    • 2023
  • The genus Paenibacillus contains a variety of biologically active compounds that have potential applications in a range of fields, including medicine, agriculture, and livestock, playing an important role in the health and economy of society. Our study focused on the bacterium SS4T (KCTC 43402T = GDMCC 1.3498T), which was characterized using a polyphasic taxonomic approach. This strain was analyzed using antiSMASH, BAGEL4, and PRISM to predict the secondary metabolites. Lassopeptide clusters were found using all three analysis methods, with the possibility of secretion. Additionally, PRISM found three biosynthetic gene clusters (BGC) and predicted the structure of the product. Genome analysis indicated that glucoamylase is present in SS4T. 16S rRNA sequence analysis showed that strain SS4T most closely resembled Paenibacillus marchantiophytorum DSM 29850T (98.22%), Paenibacillus nebraskensis JJ-59T (98.19%), and Paenibacillus aceris KCTC 13870T (98.08%). Analysis of the 16S rRNA gene sequences and Type Strain Genome Server (TYGS) analysis revealed that SS4T belongs to the genus Paenibacillus based on the results of the phylogenetic analysis. As a result of the matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) results, SS4T was determined to belong to the genus Paenibacillus. Comparing P. marchantiophytorum DSM 29850T with average nucleotide identity (ANI 78.97%) and digital DNA-DNA hybridization (dDDH 23%) revealed values that were all less than the threshold for bacterial species differentiation. The results of this study suggest that strain SS4T can be classified as a Paenibacillus andongensis species and is a novel member of the genus Paenibacillus.

A study of newly recorded genus and species for aerial cyanobacteria Wilmottia murrayi(Oscillatoriales, Cyanobacteria) in Korea (기중성 남세균, Wilmottia murrayi (Oscillatoriales, Cyanobacteria)의 국내 미기록속 및 미기록종에 대한 연구)

  • Lee, Nam-Ju;Seo, Yoseph;Ki, Jang-Seu;Lee, Ok-Min
    • Korean Journal of Environmental Biology
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    • v.37 no.3
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    • pp.260-267
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    • 2019
  • Two aerophytic cyanobacteria from the rockwall of Haje port located in Geum river, Korea, were isolated in unialgal cultures and submitted to polyphasic evaluation. The filaments of the populations presented solitary or several to many parallel arranged. The straight trichomes were not attenuated with rounded apical cell. Phylogenetic analyses based on 16S rDNA sequences indicated that these populations formed the same clade with Wilmottia murrayi and had 99% or greater DNA similarity. Through the ultrastructure of the TEM, these populations showed parietal thylakoid arrangement, which coincides with family Coleofasciculaceae. From the above results, we reported the newly recorded genus Wilmottia, and species W. murrayi in Korea.

Arthrobacter subterraneus sp. nov., Isolated from Deep Subsurface Water of the South Coast of Korea

  • Chang, Ho-Won;Bae, Jin-Woo;Nam, Young-Do;Kwon, Hyuk-Yong;Park, Ja-Ryeong;Shin, Kee-Sun;Kim, Kyoung-Ho;Quan, Zhe-Xue;Rhee, Sung-Keun;An, Kwang-Guk;Park, Yong-Ha
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1875-1879
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    • 2007
  • Strain $CH7^T$, a pale yellow-pigmented bacterium and new isolate from deep subsurface water of the South Coast of Korea, was subjected to a polyphasic taxonomic study. $CH7^T$ grew between 5 and $37^{\circ}C$, pH 5.3-10.5, and tolerated up to 13% NaCl. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain $CH7^T$ was associated with the genus Arthrobacter and phylogenetically closely related to the type strains Arthrobacter tumbae (99.4%) and Arthrobacter parietis (99.1%). However, DNA-DNA hybridization experiments revealed 2.1% and 12% between strain $CH7^T$ and Arthrobacter tumbae and Arthrobacter parietis, respectively. Thus, the phenotypic and phylogenetic differences suggested that $CH7^T$ should be placed in the genus Arthrobacter as a novel species, for which the name Arthrobacter subterraneus sp. novo is proposed. In addition, the type strain for the new species is $CH7^T$ ($=KCTC\;9997^T=DSM\;17585^T$).

Taxonomic Characterization, Evaluation of Toxigenicity, and Saccharification Capability of Aspergillus Section Flavi Isolates from Korean Traditional Wheat-Based Fermentation Starter Nuruk

  • Bal, Jyotiranjan;Yun, Suk-Hyun;Chun, Jeesun;Kim, Beom-Tae;Kim, Dae-Hyuk
    • Mycobiology
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    • v.44 no.3
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    • pp.155-161
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    • 2016
  • The most economically important species used in a wide range of fermentation industries throughout Asia belong to Aspergillus section Flavi, which are morphologically and phylogenetically indistinguishable, with a few being toxigenic and therefore a major concern. They are frequently isolated from Korean fermentation starters, such as nuruk and meju. The growing popularity of traditional Korean alcoholic beverages has led to a demand for their quality enhancement, therefore requiring selection of efficient non-toxigenic strains to assist effective fermentation. This study was performed to classify the most efficient strains of Aspergillus section Flavi isolated from various types of traditional wheat nuruk, based on a polyphasic approach involving molecular and biochemical evaluation. A total of 69 strains were isolated based on colony morphology and identified as Aspergillus oryzae/flavus based on internal transcribed spacer and calmodulin gene sequencing. Interestingly, none were toxigenic based on PCR amplification of intergenic regions of the aflatoxin cluster genes norB-cypA and the absence of aflatoxin in the culture supernatants by thin-layer chromatography analysis. Saccharification capability of the isolates, assessed through ${\alpha}-amylase$ and glucoamylase activities, revealed that two isolates, TNA24 and TNA15, showed the highest levels of activity. Although the degrees of variation in ${\alpha}-amylase$ and glucoamylase activities among the isolates were higher, there were only slight differences in acid protease activity among the isolates with two, TNA28 and TNA36, showing the highest activities. Furthermore, statistical analyses showed that ${\alpha}-amylase$ activity was positively correlated with glucoamylase activity (p < 0.001), and therefore screening for either was sufficient to predict the saccharifying capacity of the Aspergillus strain.

Photochemical assessment of maize (Zea mays L.) seedlings grown under water stress using photophenomics technique

  • Ham, Hyun Don;Kim, Tea Seong;Yoo, Sung Yung;Park, Ki Bae;Kim, Tae Wan
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.341-341
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    • 2017
  • Abiotic stress adversely affects crop growth worldwide. Drought of the major abiotic stresses have the most significant impact on all of the crop. The main objective of this study was to assess the effects of drought stress on photochemical performance and vitality of maize (Zea mays L.). The photochemical characteristics were analyzed in the context of period of drought stress during the maize growth. Drought experiment was carried out for four weeks, thereafter, the drought treated maize was re-watered. The polyphasic OJIP fluorescence transient was used to evaluate the behavior of photosystem II (PSII) and photosystem I (PSI) during the entire experiment period. In drought stress, the performance Index (PI) level was reached earlier when compared to the controls. For the screening of drought stress tolerance the drought factor index (DFI) of each variety was calculated as follow DFI= log(A) + 2log(B). All the fourteen cultivars show DFI ranged from -0.69 to 0.30, meaning less useful in selection of drought tolerant cultivars. PI and electron transport flux values of fourteen cultivars were to indicate reduction of photosynthetic performance during the early vegetative stage under drought stress. In conclusion, DFI and energy flux parameters can be used as photochemical and physiological index.

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