• The Plant Pathology Journal
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• v.22 no.3
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• pp.289-294
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• 2006

Two isolates of mucous bacteria, mc75 and pc78, were isolated from fungal culture plate as culture contaminants with an interesting swarming motility. Both isolates were identified as Pseudomonas fluorescens based on microscopy, biochemical analysis, Biolog test and DNA sequence analysis of the 16S rRNA gene. Both strains have the exactly the same 16S rRNA gene sequences, and yet their biological control activity were not identical each other. In vitro analysis of antagonistic activity of two isolates against several plant pathogenic fungi indicated that both produced diffusible and volatile antifungal compounds of unknown identities. Treatment of the bacterial culture of P. fluorescens pc78 and its culture filtrate exhibited a strong biological control activity against rice sheath blight in vivo among six plant diseases tested. More effective disease control activity was obtained from treatment of bacterial culture than that of culture filtrate. Therefore, in addition to antifungal compound and siderophore production, other traits such as biofilm formation and swarming motility on plant surface may contribute to the biological control activity of P.fluorescens pc78 and mc75.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.393-399
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• 2004

Useful genes can be Screened from various environments by construction of metagenomic DNA libraries. In this study, water samples were collected from several lakes in mid Korea, and analyzed by T-RFLP to examine diversities of the microbial communities. The crude DNAs r were extracted by the SDS-based freezing-thawing method, and then further purified using an $UltraClean^{TM}$ kit (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoR I, BamH I, and Sac II in Escherichia coli DH 10B using the pBACe3.6 vector. About 44.0 Mb of metagenomic libraries were obtained with average inserts 13-15 kb in size. The bphC genes responsible for degradation of aromatic hydrocarbons via mets-cleavage were identified from the metagenomic libraries by colony hybridization using the bphC specific sequence as a probe. The 2,3-dihydroxybiphenyl (2, 3-DHBP) dioxygenase gene (bphC ), capable of degradation of 2,3-DHBP, was cloned and its nucleotide Sequences analyzed. The genes consisted of 966 and 897 base pairs with an ATG initiation codon and a TGA termination codon. The activity of the 2,3-DHBP dioxygenase was highly expressed to 2,3-DHBP and Showed a broad substrate range to 2,3-DHBP, catechol, 3-methylcatechol and 4-methylcatechol. These results in-dicated that the bphC gene identified from the metagenomes derived from lake water might be useful in the development of a potent strain for degradation of aromatic pollutants.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.311-315
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• 2012

A novel ${\beta}$-proteobacterium, designated BXN5-$27^T$, was isolated from soil of a ginseng field of Baekdu Mountain in China, and was characterized using a polyphasic approach. The strain was Gram-staining-negative, aerobic, motile, non-spore-forming, and rod shaped. Strain BXN5-$27^T$ exhibited ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to compound Rd. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belonged to the family Comamonadaceae; it was most closely related to Ramlibacter henchirensis $TMB834^T$ and Ramlibacter tataouinensis$TTB310^T$ (96.4% and 96.3% similarity, respectively). The G+C content of the genomic DNA was 68.1%. The major menaquinone was Q-8. The major fatty acids were $C_{16:0}$, summed feature 4 (comprising $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2OH), and $C_{17:0}$ cyclo. Genomic and chemotaxonomic data supported the affiliation of strain BXN5-$27^T$ to the genus Ramlibacter. However, physiological and biochemical tests differentiated it phenotypically from the other established species of Ramlibacter. Therefore, the isolate represents a novel species, for which the name Ramlibacter ginsenosidimutans sp. nov. is proposed, with the type strain being BXN5-$27^T$ (=DSM $23480^T$ = LMG $24525^T$ = KCTC $22276^T$).

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.86-95
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• 2015

To study the halobacterial diversity at the rhizospheric soil of coastal plant native to Dokdo islands, several host plant were selected and its rhizospheric soil was sampled. Soil sample was diluted serially and pure isolation was done by sub-culture using marine agar media. 26 halophilic strains cultivable at the marine medium containig concentration of 9.0% sodium chloride were selected among total 161 isolates. Their partial 16S rRNA gene sequences extracted from genomic DNA were analyzed and partially identified. Furthermore, to identify their genetic relationship, phylogenetic tree was deduced. Total 26 strains were belongs to Firmicutes (30.8%), Gamma proteobacteria (53.8%), Bacteroidetes (7.7%), Alpha proteobacteria (7.7%), and Actinobacteria (7.7%). These results showed the specific difference from previous researches which has been reported the microbial flora of soil or sea water around the Dokdo islands. Furthermore, 4 among 26 halophilic strains grew at above 12.0% NaCl concentrated marine broth, and 2 strains Idiomarina abyssalis LM4H23 and Halomonas huangheensis AS4H13 grew at 15.0% concentration. These halophilic strains thought to overcoming the severe stress like high salt concentration or variation derived from Dokdo-specific climate and might have unknown, specific relationship with their host coastal plant native to Dokdo islands.

• Molecules and Cells
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• v.26 no.6
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• pp.536-547
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• 2008

Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.41-41
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• 2014

Pinewood nematode (PWN, Bursaphelenchus xylophilus) is a serious pathogenic worm that quickly dry pine trees to death. Recently, PWN has been devastating huge amounts of conifer trees in Korea. As a first step to explore the association and ecological roles of fungi in PWN life cycle in Korea, in this study we first isolated and indentified fungi from PWN-infested Korean pine and Japanese black pine wood sampled in Jinju, Sacheon, Pocheon, Chuncheon, Gwangju, and Hoengseong in Korea. A total of 144 fungal isolates were obtained from Japanese black pine wood and 264 fungal isolates from Korean pine wood. Their morphology and nucleotide sequences of the ITS rDNA and ♌-tubulin gene were examined for species identification. Ophiostoma ips, Botrytis anthophila, Penicillium sp., Hypocrea lixii, Trichoderma atroviride, O. galeiforme, Fusarium proliferatum were identified from Japanese black pine wood. Leptographium koreanum, L. pini-densiflorae, Ophiostoma ips, Penicillium raistrick, Trichoderma sp. were isolated from Korean pine wood. O. ips and L. koreanum were the major species on the two different PWN-infected pine tree. The cultivation of PWN on fungal mat of the identified species did some enhance PWN reproduction. The ambrosia beetle, Platypus koryoensis, is a serious pest of oak trees in Korea. In this study we investigated filamentous fungi present in the body of the beetle. Fourteen genera of filamentous fungi belonging to Ascomycota and Basidiomycota were isolated. All the obtained genera were isolated in the mitosporic state. The identified fungi were classified in 11 distinct orders including the Ascomycota (Eurotiales, Hypocreales, Microascales, Ophiostomatales, Pleosporales, and Sordiales) and Basidiomycota (Agaricales, Corticiales, Polyporales, and Russulales Xylariales). Within Ascomycota, 13 species were found. Meanwhile five species were found within Basidiomycota. The results showed the presence of diverse fungi in P. koryoensis. Among the isolated fungi, some were able to produce wood degrading enzymes. Further fungal isolation was performed with P. koryoensis infested Quercus mongolica trees sampled at Kumdan mountain in Hanam-Si, Gyeonggi province from June of 2009 to June of 2010. Penicillin spp. and Trichoderma spp. were the major species of mold fungi group. Pichia guilliermondii was the major species of mold yeast group. Raffaelea quercus-mongolicae was also isolated, but its isolation frequency was not high. Other species identified were Ambrosiella xylebori, Fusarium solani, Cryphonectria nitschke, Chaetomium globosum, and Gliocladium viride, Candida kashinagacola, C. maritima, C. vanderkliftii, Saccharomycopsis crataegensis.

• Journal of Microbiology
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• v.37 no.3
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• pp.128-135
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• 1999

Among oil-degrading microorganisms isolated from oil-polluted industrial areas, one yeast strain showed high degradation activity of aliphatic hydrocarbons. From the analyses of 18S rRNA sequences, fatty acid, coenzyme Q system, G+C content of DNA, and biochemical characteristics, the strain was identified as Yarrowia lipolytica 180. Y. lipolytica 180 degraded 94% of aliphatic hydrocarbons in minimal salts medium containing 0.2% (v/v) of Arabian light crude oil within 3 days at 25$^{\circ}C$. Optimal growth conditions for temperature, pH, NaCl concentration, and crude oil concentration were 30$^{\circ}C$, pH 5-7, 1%, and 2% (v/v), respectively. Y. lipolytica 180 reduced surface tension when cultured on hydrocarbon substrates (1%, v/v), and the measured values of the surface tension were in the range of 51 to 57 dynes/cm. Both the cell free culture broth and cell debris of Y. lipolytica 180 were capable of emulsifying 2% (v/v) crude oil by itself. They were also capable of degrading crude oil (2%). The strain showed a cell surface hydrophobicity higher than 90%, which did not require hydrocarbon substrates for its induction. These results suggest that Y. lipolytica has high oil-degrading activity through its high emulsifying activity and cell hydrophobicity, and further indicate that the cell surface is responsible for the metabolism of aliphatic hydrocarbons.

• The Plant Pathology Journal
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• v.28 no.3
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• pp.270-281
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• 2012

The bacteria B1-9 that was isolated from the rhizosphere of the green onion could promote growth of pepper, cucumber, tomato, and melon plants. In particular, pepper yield after B1-9 treatment on the seedling was increased about 3 times higher than that of control plants in a field experiment. Partial 16S rDNA sequences revealed that B1-9 belongs to the genus Pantoea ananatis. Pathogenecity tests showed non-pathogenic on kimchi cabbage, carrot, and onion. The functional characterization study demonstrated B1-9's ability to function in phosphate solubilization, sulfur oxidation, nitrogen fixation, and indole-3-acetic acid production. To trace colonization patterns of B1-9 in pepper plant tissues, we used $DRAQ5^{TM}$ fluorescent dye, which stains the DNAs of bacteria and plant cells. A large number of B1-9 cells were found on the surfaces of roots and stems as well as in guard cells. Furthermore, several colonized B1-9 cells resided in inner cortical plant cells. Treatment of rhizosphere regions with strain B1-9 can result in efficient colonization of plants and promote plant growth from the seedling to mature plant stage. In summary, strain B1-9 can be successfully applied in the pepper plantation because of its high colonization capacity in plant tissues, as well as properties that promote efficient plant growth.

• Parasites, Hosts and Diseases
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• v.58 no.1
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• pp.15-25
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• 2020

The third stage larvae (L3) of Anisakis typica were detected in 2 species of threadfin bream, Nemipterus hexodon and N. japonicus, from the Gulf of Thailand, and were morphologically and molecularly characterized. Total 100 threadfin breams, 50 Nemipterus hexodon and 50 N. japonicus, were examined with naked eyes after the opening of abdominal cavity with scissors. Almost all infected larvae remained alive and active even the fish were transported for 1-2 days. Anisakid larvae were exclusively distributed in the body cavity and rarely in the liver. The prevalence of A. typica L3 were 68.0% and 60.0% in N. hexodon and N. japonicus and their infection intensities were 3.5 and 4.2 per fish infected each. Morphological and morphometric analysis were performed by viewing specimens under both a light microscope and a scanning electron microscope. Interestingly, the protruded mucron of Anisakis typica under SEM showed a distinct cylindrical shape that differed from the cone shape of A. simplex. The protruded mucron could be used to identify A. typica L3 larvae in the future. A comparison of the ITS1-5.8S-ITS2 rDNA nucleotide sequences of these species revealed high blast scores with A. typica. Conclusively, it was confirmed that A. typica L3 are prevalent in threadfin breams from the Gulf of Thailand, and their morphological and molecular characters are something different from those of other anisakid larvae, including A. simplex and A. pegreffii.

• The Korean Journal of Mycology
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• v.48 no.3
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• pp.337-343
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• 2020

During the extensive forays for discovering the biodiversity of phytopathogenic fungi in Korea, powdery mildew-affected purple coneflowers (Echinacea purpurea) were frequently found. Since 2004, nine samples have been deposited in the Korea University Herbarium. The initial symptoms include white, evanescent mycelia and irregular patches on mature leaves. Mild symptoms in the form of discolored lesions are also noticed on the stem and petal of the affected plants. Based on the morphological characteristics of the anamorph and the phylogenetic analysis of the internal transcribed spacers (ITS) and large subunit (LSU) rDNA sequences, the causal fungus was identified as Podosphaera xanthii. This is the first report that identifies the causal agent of powdery mildew disease affecting purple coneflower in Korea.