고점도 다당류를 생산하는 갈근 내생균의 분리 및 특성

황경숙,최성현,한송이,Whang, Kyung-Sook,Choi, Seung-Hyun,Han, Song-Ih 한국미생물학회 2007 미생물학회지 Vol.43 No.4

Fifty endophytic bacteria, which produced slime around the colonies, were isolated from Pueraria roots. In particular, HDN-14, TDG-3, and TNB-3 strains, which appeared to be high viscosity producers, were selected. These strains produced high levels of polysaccharides in Puerara root medium extract. The purified polysaccharide was digested with 1N HCI and analyzed by HPLC, with glucose ($45.6{\sim}63.1%$), maltose ($14.6{\sim}23.7%$), and fructose ($17.4{\sim}23.7%$) detected as constitutive sugars. When determined by the homology relationship of the 16S rDNA sequence with the relative taxa, the HDN-14 and TNB-3 strains were closely ($99.06{\sim}99.32%$) related to the Pseudomonas $koreensis^T$ and Pseudomonas $jessenii^T$, while TDG-3 were closely ($99.48{\sim}99.74%$) related to Pseudomonas $plecoglossicida^T$, Pseudomonas $mosselii^T$, and Pseudomonas $monteilii^T$. The major cellular Pseudomonas acids are $3OH-C_{10:0}$, $2OH-C_{12:0}$, $3OH-C_{12:0}$, and $3OH-C_{12:1}$, with these strains being further differentiated in species belonging to the genus Pseudomonas.

축산폐기물의 퇴비화 과정중 미생물상의 변동

黃敬淑(Kyung-Sook Whang),張基運(Ki-Woon Chang) 한국토양비료학회 1996 한국토양비료학회지 Vol.29 No.3

송이 자생군락 토양 내 난배양성 세균군집의 계통학적 특성

김윤지,황경숙,Kim, Yun-Ji,Whang, Kyung-Sook 한국미생물학회 2007 미생물학회지 Vol.43 No.3

The CFDA (6-carboxyfluorescein diacetate) direct viable count method and plate count (PC) method using conventional nutrient broth (NB) medium and $10^{-2}$ diluted NB (DNB) medium were applied to samples collected from Mt. Yongdoo In Andong, in an effect to determine the number of living bacteria pine mushroom forest soil. The number of living bacteria determined via plate count in NB medium comprised $5{\sim}8%$ of the CFDA direct viable count, and the bacteria in the DNB medium comprised $40{\sim}47%$. This result indicated that viable but nonculturable (VBNC) bacteria existed in the pine mushroom forest soil at a high percentage. The phylogenetic characteristics of the VBNC bacterial populations in the samples of pine mushroom (Tricholoma matsutake) forest soil were analyzed via the direct extraction of DNA and 16S rDNA-ARDRA. The 115 clones from pine mushroom forest soil were clustered into 31 different RFLP phylotypes by ARDRA. Based on the 16S rDNA sequences, the 31 ARDRA clusters were classified into 6 phylogenetic groups: ${\alpha}-,\;{\beta}-,\;{\gamma}-Proteobacteria$, Acidobacteria, Actinobacteria and Firmicutes. Among these bacterial populations, approximately 85% were classified as members of phylum Acidobacteria. The Acidobacteria phylum was shown to exist abundantly in the pine mushroom forest soil.

항진균활성 violacein 색소를 생산하는 Collimonas sp. DEC-B5 균주의 분리 및 특성

이예림,황경숙,Lee, Ye-Rim,Mitchell, Robert J.,Whang, Kyung-Sook 한국미생물학회 2016 미생물학회지 Vol.52 No.2

Forty-nine pigments were extracted from the collections of 106 pigment producing bacteria from the plant rhizosphere soil. Antibacterial activity test was performed in the subjects of the extracted pigments with plant pathogenic bacteria including Xanthomonas axonopodis and Xanthomonas campestris, and with plant pathogenic fungi including Botrytis cinerea, Colletotrichum acutatum, and Fusarium oxysporum. The yellow pigment by Chryseobacterium sp. RBR9 and the red pigment by of Methylobacterium sp. RI13 showed the antibacterial activities against Xanthomonas axonopodis and Xanthomonas campestris. The violet pigment by Collimonas sp. DEC-B5 showed the antibacterial activity as well as the antifungal activities against Botrytis cinerea and Fusarium oxysporum. Especially, the violet pigment inhibited the growth of Botrytis cinerea more than 65% at MIC $20{\mu}M$. Upon the HPLC analysis result for the isolation of pigment with antifungal activity, violacein (91.6%) and deoxyviolacein (8.4%) were isolated for the pigment by Collimonas sp. DEC-B5. The production amount of the pigment was increased more than 10 times higher when D-mannitol 1.5% and yeast extract 0.2% were added as the nitrogen source to SCB medium. This study suggests that produced violacein by Collimonas sp. DEC-B5 will be effective to control strawberry gray-mold rot fungi by its preventive activity.

무당벌레 소화기관으로부터 산성 단백질 분해효소를 생산하는 Staphylococcus sp. CB2-3의 분리 및 특성

김세종,황경숙,Kim, Se-Jong,Whang, Kyung-Sook 한국미생물학회 2011 미생물학회지 Vol.47 No.3

Six protein-degrading bacteria were isolated from digestive organ of Harmonia axyridis. These isolates were categorized as Staphylococcus sciuri subsp. sciuri (3 strains), Bacillus subtilis (1 strain), and Bacillus thuringiensis (2 strains) by 16S rRNA gene sequence analysis. The Staphylococcus sp. CB2-3 was selected as a protease-producing bacterium which showed the highest protease activity of 58.5 U/ml at the pH 5.0 medium. The optimal pH and temperature of protease activity were pH 5.0 and $40^{\circ}C$, respectively. This acid protease had a relatively high stability of 80% between $30-50^{\circ}C$ at broad temperature range. The opimal medium compositions of carbon, nitrogen and mineral source for cell growth and protease activity were investigated. When sorbitol (0.5%) was used as carbon source, enzyme activity was increased about 2 times than that of the basal medium. When skim milk (0.5%) was used as nitrogen source, activity was increased about 2.5 times than that of the control. Cell growth and enzyme activity were increased by mineral source such as KCl, $K_2HPO_4$, $FeSO_4$, but was completely inhibited by divalent ions such as $Co^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Cu^{2+}$.

대나무 산림토양으로부터 수집한 Streptomyces 속 방선균의 계통학적 다양성

이효진,황경숙,Lee, Hyo-Jin,Whang, Kyung-Sook 한국미생물학회 2010 미생물학회지 Vol.46 No.3

To investigate a quantitative evaluation of the actinobacteria, we have collected samples from various kinds of bamboo forest soil. Each different layers contained $2.7{\times}10^6-2.7{\times}10^8$ CFU/g of actinobacteria which was the highest in litter layers of Sasa boreali forest soil. We obtained 330 actinobacteria from different layers of bamboo forest soil; litter (100 strains), humus (70 strains), and rhizosphere soil (160 strains). Based on the colony morphology (aerial mycelium, substrate mycelium, and soluble pigment), isolates were divided into thirty-six groups and we selected 50 representative isolates. 16S rRNA gene sequence analysis showed Streptomyces was major actinobacteria (94%) and they were categorized as cluster I (2 strains), II (35 strains), III (6 strains), and IV (7 strains), respectively. The diversity index of 50 Streptomyces collected from the bamboo forest soil was calculated with the Shannon-Wiener method. Bamboo litter showed higher diversity index level of 3.33 than that of humus and rhizosphere soil. Also, antibiotic activities of our isolates were investigated against Botrytis cinerea, Xanthomonas campestris, Xanthomonas axonopodis pv. vesicatoria, and Bacillus cereus and found in 74, 16, 25, and 24 strains, respectively.

송이 자실체 기저부 토양으로부터 항균활성을 가지는 KM1-15 균주의 분리 및 분류학적 특성

김윤지,황경숙,Kim, Yun-Ji,Whang, Kyung-Sook 한국미생물학회 2008 미생물학회지 Vol.44 No.1

송이 자실체 기저부 토양으로부터 분리한 268균주를 대상으로 식물병원성 곰팡이 7균주(Alternaria panax, Botrytis cinerea, Colletotrichum gloeosprioides, Fusarium oxysporum, Phytopthora capsici, Pythium ultimum, Rizoctonia solani)에 대해 항균활성을 검토한 결과 인삼점무늬병원균(Alternaria panax)과 인삼탄저병원균(Colletotrichum gloeosprioides)에 강한 항균활성을 나타내는 KM1-15 균주가 선발되었다. KM1-15 균주의 16S rDNA 염기서열(1,311 bp)을 결정하여 계통학적 위치를 검토한 결과, Bacillus $koguryoae^T$ (AY904033)와 99.6%의 상동성을 나타내었다. API 50CHE Kit를 사용하여 당 분해능에 대해 Bacillus $koguryoae^T$ (AY904033)와 비교 검토한 결과 KM1-15 균주는 L-arabinose, cellobiose, inulin, D-turanose 양성반응을 나타내는 생리학적 특성이 확인되었다. 주요 지방산으로는 15:0 anteiso (35.78%), 17:0 anteiso (17.97%)를 함유하였으며, 13:0 iso 3-OH, 14:0 iso3-OH, 15:0 iso 3-OH, 그리고 17:0 iso 3-OH와 같은 다양한 분지형 지방산을 함유하는 특성을 나타내었다. 퀴논종으로는 MK-7 (100%)을 가지고, G+C함량은 43.7%를 나타내었다. 또한 protein과 starch에 대한 가수분해능이 탁월하여 KM1-15 균주는 병해충 방제제 및 토양개량제로 매우 유용하게 활용될 것으로 사료된다.

광합성(光合成) 세균(細菌) T-20주(株)의 동정(同定)

현문식,황경숙,신관철,Hyun, Moon Sik,Whang, Kyung Sook,Shin, Gwan Chull 충남대학교 농업과학연구소 1992 Korean Journal of Agricultural Science Vol.19 No.1

Phototrophic bacteria T-20 investigated in the present report was isolated from Thailand oil, Cells of the bacterium were rod-shaped. measured $0.6{\sim}0.7{\mu}m$ wide, and $2.3{\mu}m$ long. A color of cell suspension was red, and the long wavelength absorption maxima of the bacteriochlorophyll measured in 870nm. Substrates utilized as carbon sources: citrate, aspirate, glutamate, and fructose: unique for this strain is the liquefaction of gelatine, Ubiquinone Q-8 was also found. Rhodocyclus gelatinosus, which was reported in Bergey's Manual of Systematics Bacteriology, was corresponded to T-20.

DNA 직접추출법에 따른 산림토양 부식층 내 세균군집의 계통학적 다양성 비교

손희성,한송이,황경숙,Son, Hee-Seong,Han, Song-Ih,Whang, Kyung-Sook 한국미생물학회 2007 미생물학회지 Vol.43 No.3

The principal objective of this study was to analyze 16S rDNA-ARDRA of the humus forest soil via an improved manual method and an ISOIL kit on the basis of the UPGMA clustering of the 16S rDNA combined profile, 44 ARDRA clusters of 76 clones via the ISOIL kit method and 45 ARDRA clusters of 136 clones via the improved manual method. On the basis of the 16S rDNA sequences, 44 clones from the ARDRA clusters by the ISOIL kit were classified into 3 phyla : ${\alpha}-,\;{\beta}-,\;{\gamma}-,\;{\delta}-Proteobacteria$, Acidobacteria and Actinobacteria. Using the improved manual method, the specimens were classified into 6 phyla : the ${\alpha}-,\;{\beta}-,\;{\gamma}-,\;{\delta}-Proteobacteria$, Acidobacteria, Bacteroides, Verrucomicrobia, Planctomycetes and Gemmatomonadetes. As a result, the modified manual method indicated greater phylogenetic diversity than was detected by the ISOIL kit. Approximately 40 percent of the total clones were identified as ${\alpha}-Proteobacteria$ and 30 percent of the total clones were ${\gamma}-Proteobacteria$ and assigned to dominant phylogenetic groups using the ISOIL kit. Using the modified manual method, 41 percent of the total clones were identified as Acidobacteria and 28 percent of total clones were identified as ${\alpha}-proteobacteria$ and assigned to dominant phylogenetic groups.

고추 세균성 점무늬병원균(Xanthomonas axonopodis pv. vesicatoria)의 항균활성 Streptomyces sp. JR-24 균주의 분리 및 분류학적 특성

한송이,이효진,황경숙,Han, Song-Ih,Lee, Hyo-Jin,Whang, Kyung-Sook 한국미생물학회 2010 미생물학회지 Vol.46 No.4

Fifty Actinobacteria strains were isolated from rhizosphere soil of Sasa borealis. In the course of screening for antibacterial activity against bacterial leaf spot of pepper (Xanthomonas axonopodis pv. vesicatoria) of isolates, 12 isolates showed strong antibiotic activity. Basis on the 16S rRNA gene sequence, they were belonging to Streptomyces cluster II. Strain JR-24 exhibited strong antibiotic activity against X. axonopodis pv. vesicatoria, had a minimum inhibitory concentration of 10 ${\mu}l$/disc. The strain JR-24 was most closely related to Streptomyces galbus $DSM40089^T$ (98.1%), Streptomyces longwoodensis $LMG20096^T$ (98%) and Streptomyces capoamus $JCM4734^T$ (97.8%). When assayed with the API 20NE and 50 CHE kit, it is positive for utilization of L-arabinose, D-fructose, D-glucose, D-galactose and hydrolysis of gelatin, protein, starch. The strains contained iso-$C_{14:0}$ (25.93%), iso-$C_{15:0}$ (10.13%), anteiso-$C_{15:0}$ (19.29%) and iso-$C_{16:0}$ (20.35%) as major fatty acids and MK-9 (H4), MK-9 (H6), and MK-9 (H8) as the isoprenoid quinone. Strain JR-24 was suggested new species of genus Streptomyces by nearest neighbors of genotypic relationships and phenotypic characterization. This study was important to microbial resources investigation for environment-friendly agriculture.