• Title/Summary/Keyword: root isolates

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Characterization of Vancomycin Resistant Enterococci and Drug Ligand Interaction between vanA of E. faecalis with the Bio-Compounds from Aegles marmelos

  • Jayavarsha V;Smiline Girija A.S;Shoba Gunasekaran;Vijayashree Priyadharsini J
    • Journal of Pharmacopuncture
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    • v.26 no.3
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    • pp.247-256
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    • 2023
  • Objectives: Enterococcus faecalis is a gram positive diplococci, highly versatile and a normal commensal of the gut microbiome. Resistance to vancomycin is a serious issue in various health-care setting exhibited by vancomycin resistant Enterococci (VRE) due to the alteration in the peptidoglycan synthesis pathway. This study is thus aimed to detect the VRE from the patients with root caries from the clinical isolates of E. faecalis and to evaluate the in-silico interactions between vanA and the Aegles marmelos bio-compounds. Methods: E. faecalis was phenotypically characterized from 20 root caries samples and the frequency of vanA and vanB genes was detected by polymerase chain reaction (PCR). Further crude methanolic extracts from the dried leaves of A. marmelos was assessed for its antimicrobial activity. This is followed by the selection of five A. marmelos bio-compounds for the computational approach towards the drug ligand interactions. Results: 12 strains (60%) of E. faecalis was identified from the root caries samples and vanA was detected from two strains (16%). Both the stains showed the presence of vanA and none of the strains possessed vanB. Crude extract of A. marmelos showed promising antibacterial activity against the VRE strains. In-silico analysis of the A. marmelos biocompounds revealed Imperatonin as the best compound with high docking energy (-8.11) and hydrogen bonds with < 140 TPSA (Topological polar surface area) and zero violations. Conclusion: The present study records the VRE strains among the root caries with imperatorin from A. marmelos as a promising drug candidate. However the study requires further experimentation and validation.

Crown Gall of Chrosanthemum Caused by Agrobacterium rubi and A. tumefaciens (Agrobacterium rubi와 A. tumefaciens에 의한 국화 뿌리혹병)

  • Lee, Young-Kee;Park, Kyung-Seok;Hwang, Hye-Kyung;Hwang, Tae-Ho;Kim, Jin-Young;Lee, Jae-Kook;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.12 no.3
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    • pp.197-204
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    • 2006
  • Crown gall on lower stem and root of chrysanthemum(Dendranthema grandiflorum Kitamura) was observed at Hwasung and Gumi in 2001 and 2004, respectively. Tumors were semi-round with rough surface texture of dark brown color. Nine isolates inducing gall formation on lower stem of chrysanthemum among twenty isolates from the tumor tissues were characterized. Their colonies were convex, glistening, circular with an entire edge and whitish or tannish cream in color on potato dextrose agar supplemented with 0.5% $CaCO_3$. The virulent isolates were rod-shaped with peritrichous flagellae, gram negative, aerobic and growing on D1M agar. Among nine virulent isolates, one isolate was identified as Agrobacterium rubi and eight isolates were A. tumefaciens based on biochemical and physiological characteristics, fatty acid profiles and substrate utilization patterns. A. tumefaciens had strong pathogenicity and broad host range compared with A. rubi. This is the first report on crown gall of chrysanthemum in Korea. To our knowledge, crown gall of chrysanthemum caused by A. rubi is first report in this study worldwide.

Selection of Ectomycorrhizal Isolates of Tricholoma matsutake and T. magnivelare for Inoculation on Seedlings of Pinus densiflora In Vitro (소나무 유묘에서 송이 외생균근 형성 균주의 선발)

  • Ka, Kang-Hyeon;Park, Hyun;Hur, Tae-Chul;Bak, Won-Chull
    • The Korean Journal of Mycology
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    • v.36 no.2
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    • pp.148-152
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    • 2008
  • We inoculated hypal suspension of Tricholoma matsutake and T. magnivelare were examined on Pinus densiflora seedlings grown in a granite soil substrate with 1/2 PDMP (12 g/l potato dextrose broth, 1.5 g/l malt extract, and 0.5 g/l peptone) medium. Four months after inoculation, the pine seedlings were examined for infection rate, matsutake aroma, and Hartig-net formation. The roots of pine seedling formed ectomycorrhizal roots in the 9 isolates from 12 isolates of T. matsutake and T. magnivelare. However, the seedlings showed different ectomycorrhizae forming rates among the 9 isolates. While matsutake aroma was confirmed from the ectomycorrhizal seedlings, the pine seedling contaminated by bacteria or fungi did not form matsutake ectomycorrhizae with sickening smell. Thus, the aroma was chosen as a good way for the verification of mycorrhizal infection. At the early stage, the mycorrhizal roots showed unramified and branched types without root hair. They also showed thin mantle layers, Hartig-nets, and turned into black color at later stage. Among the examined strains, that of Yecheon isolated in 1995 showed the best infection rate, which indicated that we need to pay attention to the selection of isolates for better result.

Diversity and Pathogenic Characteristics of Fusarium Species isolated from Wilted Soybeans in Korea (콩 시들음병에 관여하는 Fusarium균의 다양성 및 병원학적 특성)

  • Choi, Hyo-Won;Kim, Seungnoh;Hong, Sung Kee
    • The Korean Journal of Mycology
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    • v.48 no.3
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    • pp.297-312
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    • 2020
  • Wilted soybean plants were collected from soybeans cultivation fields in Korea from 2014 to 2016. Fusarium spp., Colletotrichum spp., Rhizoctonia spp., Macrophomina sp., Phytophthora spp., and Calonectria ilicicola were obtained from the infected samples. Out of these, Fusarium spp. were the dominant species (79.1%). In total, 53 isolates were identified as F. solani species complex, F. oxysporum species complex, F. graminearum species complex, and F. fujikuroi species complex based on mycological characteristics. Sequence typing analysis was conducted using translation elongation factor 1 alpha (TEF) to confirm the identification of isolates. All isolates were identified as F. solani, F. oxysporum, F. commune, F. asiaticum, and F. fujikuroi based on phylogenetic analysis of TEF sequences. Pathogenicity of 44 isolates was tested on three cultivars of soybean using the root dip inoculation method. Out of 5 Fusarium species, only F. asiaticum could not cause the symptoms or be weak. Ten isolates were selected based on pathogenic characters and species identification to investigate the host range and screen soybean cultivars for resistance. Fusarium solani, F. oxysporum, and F. commune were aggressive only to soybean, and F. fujikuroi was aggressive to kidney bean, yellow cowpea, black cowpea, adzuki bean as well as soybean. All 13 Korean soybean cultivars were susceptible to F. commune and F. fujikuroi. Out of 13 cultivars, cv. Janggi, cv. Poongsannamul, and cv. Socheongja were resistant to Fusarium wilt, while cv. Hwanggeumol and Chamol were susceptible to Fusarium wilt.

Conversion of Ginsenoside Rb1 and Taxonomical Characterization of Stenotrophomonas sp. 4KR4 from Ginseng Rhizosphere Soil (인삼 근권 토양에서 분리한 Stenotrophomonas sp. 4KR4의 Ginsenoside Rb1 전환능 및 분류학적 특성)

  • Jeon, In-Hwa;Cho, Geon-Yeong;Han, Song-Ih;Yoo, Sun Kyun;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.369-376
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    • 2013
  • We isolated the ${\beta}$-glucosidase producing bacteria (BGB) in ginseng root system (rhizosphere soil, rhizoplane, inside of root). Phylogenetic analysis of the 28 BGB based on the 16S rRNA gene sequences, BGB from rhizosphere soil belong to genus Stenotrophomonas (3 strains), Bacillus (1 strain), and Pseudoxanthomonas (1 strain). BGB isolates from rhizoplane were Stenotrophomonas (16 strains), Streptomyces (1 strain) and Microbacterium (1 strain). BGB from inside of root were categorized into Stenotrophomonas (3 strains) and Lysobacter (2 strains). Especially, Stenotrophomonas comprised the largest portion (approximately 90%) of total isolates and Stenotrophomonas was a dominant group of the ${\beta}$-glucosidase producing bacteria. We selected strain 4KR4, which had high ${\beta}$-glucosidase activity (108.17 unit), could transform ginsenoside Rb1 into Rd, Rg3, and Rh2 ginsenosides. In determining its relationship on the basis of 16S rRNA sequence, 4KR4 strain was most closely related to Stenotrophomonas rhizophila e-$p10^T$ (AJ293463) (99.62%). Therefore, on the basis of these polyphasic taxonomic evidence, the ginsenoside Rb1 converting bacteria 4KR4 was identified as Stenotrophomonas sp. 4KR4 (=KACC 17635).

Effect of Bacillus spp. Having Gelatin Decomposing Activity on Root-knot Nematode, Meloidogyne incognita on Pepper (젤라틴 분해력이 있는 Bacillus균의 고추 고구마뿌리혹선충에 미치는 영향)

  • Kim, Jin Han;Park, Chan Sun;Kim, Young-Ho;Kim, Yu Ri;Lee, Seung Woong;Rho, Mun Chual
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.375-379
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    • 2016
  • This study investigated the decomposing activities on gelatin and egg sac of root-knot nematode (Meloidogyne incognita). Ten Bacillus isolates (KRB-1~10) isolated from the soils of Mt. Naejang. Among them, KRB-5, 9, and 10 showed decomposing activities, and identified as Bacillus subtilis KRB-5, Bacillus amyloliquefaciens KRB-9, and KRB-10 by 16S rRNA sequence analysis, respectively. Under pot experiments using pepper, 100-fold diluted culture broth of three isolates reduced the number of egg sac on roots collected 60 days and 90 days and increased the growth of aerial part compared with the group only treated with Meloidogyne incognita. Especially, the group treated B. subtilis KRB-5 was superior in the growth of pepper. These results suggest that the B. subtilis KRB-5 can be used to control the root-knot nematode on pepper.

Effect of Carbon and Nitrogen Sources on the Mycelial Growth and Sporulation of Cylindrocarpon destructans Causing Root Rot of Panax ginseng (인삼 뿌리썩음병균 Cylindrocarpon destructans의 균사생육과 포자형성에 미치는 탄소원가 질소원의 영향)

  • 조대휘;유연현;오승환;이호자
    • Korean Journal Plant Pathology
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    • v.13 no.1
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    • pp.30-36
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    • 1997
  • The effects of carbon and nitrogen sources on the mycelial growth and sporulation of microconidia and chlamydospores of five isolates of Cylindrocarpon destructans (Zinssm.) Scholten causing root rot of Panax ginseng were studied. For the carbon sources, fructose, glucose, maltose, and sucrose in Czapek-Dox broth showed good mycelial growth of 178∼201 mg in dry weight compared with 64 mg of the control. The best carbon sources tested for conidial formation were sucrose and maltose with 2.75 and 3.03 log conidia/ml, respectively. For the nitrogen sources, aspartic acid, NaNO3, KNO3, arginine, threonine, and leucine increased mycelial growth of the fungi to 208∼231 mg in dry weight without significant difference (p=0.05) among them. Meanwhile the growth with cystine was poor (26.3 mg dry weight), and no conidium and chlamydospore were formed. Maximum microconidial formation was observed in the media with NaNO3 and KNO3 as 3.37 and 3.35 log conidia/ml, and for the chlamydospore formation the (NH4)2SO4-containing medium and the nitrogen-absent medium were the best as 3.40 and 3.57 log chlamydospores/ml, respectively. No conidium was found in the medium without nitrogen sources, in which chlamydospore formation increased 6 times more than in the nitrogen-amended medium. However, deletion of carbon source in the medium did not affect on the formation of conidia and chlamydospores of C. destructans.

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Occurrence of Eggplant Wilt Caused by Verticillium dahliae

  • Kim, Sung-Kee;Kim, Ki-Woo;Park, Eun-Woo;Hong, Soon-Sung
    • The Plant Pathology Journal
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    • v.16 no.3
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    • pp.156-161
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    • 2000
  • A wilt disease occurred on greenhouse-grown eggplants at Yeojoo, Korea in 1997. The wilted eggplants had leaves with gradual yellowing, interveinal necrosis, and marginal crinkling. Vascular tissues of diseased stems were discolored, turned black, and microsclerotia developed at the base of stems. The disease progressed from lower parts of the plants upward. Fungal isolates from discolored vascular tissues were initially whitish to cream color on potato-dextrose agar (PDA) plate, which later turned black due to the formation of microsclerotia. Conidiophores were erect, hyaline, verticillately branched, and had 3 or 4 phialides arising at each node. Phialides were hyaline, arranged in whorls, and measured as 17.5-32.5 x 2-3$\mu\textrm{m}$. Conidia were hyaline, ellipsoidal to sub-cylindrical, mainly one-celled, and measured as 5-8.8 x 2-4$\mu\textrm{m}$. Conidia were borne in small clusters at the tips of phialides. Microsclerotia formed on PDA plates, and consisted of globular cells that formed irregular masses of various shapes. Chlamydospores were absent. Based on these cultural and morphological characteristics, the fungus was identified as Verticillium dahliae Klebahn. Pathogenicity tests by root cutting, root dipping or soil drenching resulted in similar symptoms observed in the naturally infected eggplants. This is the first report on occurrence of Verticillium wilt of eggplant in Korea.

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Diversity and Saline Resistance of Endophytic Fungi Associated with Pinus thunbergii in Coastal Shelterbelts of Korea

  • Min, Young Ju;Park, Myung Soo;Fong, Jonathan J.;Quan, Ying;Jung, Sungcheol;Lim, Young Woon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.324-333
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    • 2014
  • The Black Pine, Pinus thunbergii, is widely distributed along the eastern coast of Korea and its importance as a shelterbelt was highlighted after tsunamis in Indonesia and Japan. The root endophytic diversity of P. thunbergii was investigated in three coastal regions; Goseong, Uljin, and Busan. Fungi were isolated from the root tips, and growth rates of pure cultures were measured and compared between PDA with and without 3% NaCl to determine their saline resistance. A total of 259 isolates were divided into 136 morphotypes, of which internal transcribed spacer region sequences identified 58 species. Representatives of each major fungi phylum were present: 44 Ascomycota, 8 Zygomycota, and 6 Basidiomycota. Eighteen species exhibited saline resistance, many of which were Penicillium and Trichoderma species. Shoreline habitats harbored higher saline-tolerant endophytic diversity compared with inland sites. This investigation indicates that endophytes of P. thunbergii living closer to the coast may have higher resistance to salinity and potentially have specific relationships with P. thunbergii.

Monoamine Oxidase Inhibitory Flavonoids from the Root Bark of Cudrania tricuspidata

  • Han, Xiang-Hua;Hwang, Ji-Hye;Hong, Seong-Su;Choe, Sang-Gil;Lee, Chul;Lee, Moon-Soon;Lee, Dong-Ho;Lee, Myung-Koo;Lee, Mi-Kyeong;Hwang, Bang-Yeon
    • Natural Product Sciences
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    • v.16 no.2
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    • pp.75-79
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    • 2010
  • Two new benzylated flavonoids, 5,7,4'-trihydroxy-6-p-hydroxybenzylflavanone (1) and 5,7,4'-trihydroxy-6,8-di-p-hydroxybenzylflavanone (2) together with six known flavonoids, kaempferol (3), artocarpesin (4), cycloartocarpesin (5), cudraflavone D (6), gericudranin E (7), and leachianone G (8) have been isolated from the root bark of Cudrania tricuspidata. The structures of 1 and 2 were characterized based on spectroscopic data including 1D- and 2D-NMR. All the isolates were evaluated for their inhibitory effects of monoamine oxidase (MAO). Among them, kaempferol (3), artocarpesin (4), and cudraflavone D (6) showed moderate inhibitory effects with $IC_{50}$ values of 82.3, 30.8, and $71.8\;{\mu}M$, respectively.