• Title/Summary/Keyword: 탄저균

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Microbial Forensics: Comparison of MLVA Results According to NGS Methods, and Forensic DNA Analysis Using MLVA (미생물법의학: 차세대염기서열분석 방법에 따른 MLVA 결과 비교 및 이를 활용한 DNA 감식)

  • Hyeongseok Yun;Seungho Lee;Seunghyun Lim;Daesang Lee;Sehun Gu;Jungeun Kim;Juhwan Jeong;Seongjoo Kim;Gyeunghaeng Hur;Donghyun Song
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.4
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    • pp.507-515
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    • 2024
  • Microbial forensics is a scientific discipline for analyzing evidence related to biological crimes by identifying the origin of microorganisms. Multiple locus variable number tandem repeat analysis(MLVA) is one of the microbiological analysis methods used to specify subtypes within a species based on the number of tandem repeat in the genome, and advances in next generation sequencing(NGS) technology have enabled in silico anlysis of full-length whole genome sequences. In this paper, we analyzed unknown samples provided by Robert Koch Institute(RKI) through The United Nations Secretary-General's Mechanism(UNSGM)'s external quality assessment exercise(EQAE) project, which we officially participated in 2023. We confirmed that the 3 unknown samples were B. anthracis through nucleic acid isolation and genetic sequence analysis studies. MLVA results on 32 loci of B. anthracis were analysed by using genome sequences obtained from NGS(NextSeq and MinION) and Sanger sequencing. The MLVA typing using short-reads based NGS platform(NextSeq) showed a high probability of causing assembly error when a size of the tandem repeats was grater than 200 bp, while long-reads based NGS platform(MinION) showed higher accuracy than NextSeq, although insertion and deletion was observed. We also showed hybrid assembly can correct most indel error caused by MinION. Based on the MLVA results, genetic identification was performed compared to the 2,975 published MLVA databases of B. anthracis, and MLVA results of 10 strains were identical with 3 unkonwn samples. As a result of whole genome alignment of the 10 strains and 3 unknown samples, all samples were identified as B. anthracis strain A4564 which is associated with injectional anthrax isolates in heroin users.

In vitro Test of Mycelial Growth Inhibition of 5 Fungi Pathogenic to Strawberries by Ultraviolet-C (UV-C) Irradiation (자외선(UV-C) 조사에 의한 딸기병원균의 균사생장억제)

  • Kim, Seon Ae;Ahn, Soon-Young;Oh, Wook;Yun, Hae Keun
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.634-637
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    • 2012
  • In strawberry production, among others, the high incidence of diseases by pathogenic fungi resulting in the reduction of fruit yield and quality requires the development of eco-friendly management systems rather than chemical sprays to control them. The diameter of colonies grown in media at $25^{\circ}C$ for 5 days was measured to evaluate the in vitro inhibition of mycelial growth of 5 pathogenic fungi by irradiation with ultraviolet (UV-C, 264 nm). The mycelial growth of 5 pathogenic fungi was inhibited in potato dextrose agar (PDA) by the irradiation of UV-C for 1 hour a day, and was dramatically inhibited by the irradiation of UV-C for 9-12 h a day. The irradiation of UV-C for 9-12 h a day inhibited completely the growth of the late blight pathogen, Phytophthora cactorum. The irradiation distance of 40 to 50 cm was effective for the inhibition of mycelial growth of fungi. The mycelial growth of fungi without pre-incubation was inhibited strongly by UV-C irradiation compared to fungi pre-incubated for 2 days without light. The mycelia growth of Colletotrichum gloeosprioides and Fusarium oxysporum was inhibited strongly by UV-C irradiation in vegetable 8 juice agar compared to PDA.

Isolation and characterization of antifungal violacein producing bacterium Collimonas sp. DEC-B5 (항진균활성 violacein 색소를 생산하는 Collimonas sp. DEC-B5 균주의 분리 및 특성)

  • Lee, Ye-Rim;Mitchell, Robert J.;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.212-219
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    • 2016
  • 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.

Antifungfal Activity Against Plant Pathogenic Fungi on Insect Enterobacteriaceae (식물병원성 곰팡이에 대한 곤충장내세균의 항균활성)

  • Oh, San Na;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.19 no.1
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    • pp.71-79
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    • 2015
  • In order to investigating the effects of antifungal activity of intestinal bacteria obtained from insect, it was identified these bacteria isolated from the gut. In this result, total 49 isolates of intestinal bacteria were identified from 10 kinds of insect species. It was that 4 isolates including Cedecea sp. from Nesidiocoris tenuis, 3 isolates including Enterobacter sp. from Odontotaenius disjunctus, 4 isolates including Acinetobacter sp. from Reticulitermes speratus, 4 isolates including Clavibacter sp. from Riptortus clavatus, 11 isolates including Bacillus sp. from Lema decempunctata, 3 isolates including Enterococcus sp. from Henosepilachna vigintioctopunctata 2 isolates including Staphylococccus sp. from Harmonia axyridis, 5 isolates including Enterobacter asburiae from Popillia mutans, 7 isolates including Aeromonas sp. from Hydrophilus acuminatus, and 7 isolates including Brucella sp. from Anomala octiescostata. In order to investigating antifungal activity against plant-pathogenic fungi, Altanaria solani, Colletotrichum gloeosporioides, Botrytis cinerea, Fusarium oxysporum, Phytophthora capsici, Rhizoctonia solani and Selerotinia sclerotiorum were dual cultured with each 49 gut enterobacteriaceae. As these results showed that many isolates have the antifungal activities including 26 isolates against A. solani, 6 isolates against B. cinerea, 13 isolates against C. gloeosporioides, 11 isolates against F. oxysporum, 17 isolates P. capsici, 2 isolates against R. solani and 2 isolates against S. sclerotiorum. Pseudomonas aeruginosa was showed strong antifungal activity against all of tested plant pathogens. It might be taken a potential for application against plant-pathogenic fungi with useful control agent.

Seedling Growth of Red Pepper and Antagonism on Alternaria alternata Causing Black Rot by Bacillus subtilis J-24 isolated from Red Pepper Rhizosphere (고추 근권에서 분리한 Bacillus subtilis J-24의 검은 곰팡이병원균 Alternaria alternata에 대한 길항력 및 고추의 초기 생육에 미치는 영향)

  • Joo, Gil-Jae;Kim, Hak-Yoon;Hur, Sang-Sun;Woo, Churl-Joo;Rhee, In-Koo
    • Current Research on Agriculture and Life Sciences
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    • v.17
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    • pp.7-13
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    • 1999
  • Thirty bacteria were isolated from the red pepper rhizosphere. The isolates were screened for antagonism to Alternaria alteranta causing red pepper black rot. Antagonistic bacterium No. J-24 was selected among the isolated bacteria and was identified as Bacillus subtilis based on morphological and physiological characteristics and MIDI system. B. subtilis J-24 showed antifungal activities against A. alternata(inhibition percentage, 99%), Botrytis cinerea, Phytophthora capsici, Pythium ultimum, Colletotrichum gloeosporioides, Stemphylium botryosum. The growth of red pepper seedling was promoted as compared to control when the microbial inoculants was mixed in bed soil. In the mixed microbial inoculants bed soil, the leaf area of red pepper was increased of 15 percent, the hypocotyl weight 12 percent, the root length 12 percent, total dry weight 13 percent as compared to those grown in the general bed soil.

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Identification of Glomerella cingulata from Dracaena sanderiana (Dracaena sanderiana로부터 Glomerella cingulata의 동정)

  • Seo, Il-Gyo;Shim, Chang-Ki;Kim, Dong-Gil;Bae, Dong-Won;Kim, Hee-Kyu
    • Research in Plant Disease
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    • v.8 no.4
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    • pp.224-227
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    • 2002
  • From the necrotic stem of aquaculture Dracaena for living room decoration, we observed conidia on acervuli with occasional setae and perithecia in vivo. Morphological characters were; acervuli, dark brown and 300-500 ${\mu}{\textrm}{m}$ in diameter; conidia, hyaline, aseptate, cylindrical to slightly ellipsoid with rounded apex and narrow truncate base on phialidic conidiophores, 12.5-17.5 ${\mu}{\textrm}{m}$; setae, variable in length 1-4 septate, slightly swollen at the base and tapered to the apex, 200 ${\mu}{\textrm}{m}$; appressoria, dark brown ovate to obovate irregular 5-12.5 ${\mu}{\textrm}{m}$, perithecia on decayed stem, globose, dark brown and black 85-300 ${\mu}{\textrm}{m}$; asci 8 spored, clavate to cylindrical; 50-62.5$\times$8-10 ${\mu}{\textrm}{m}$; ascospores, oval to fusiform, sometimes slightly curved, aseptate, hyaline. Above characteristics were also confirmed in vitro. Colletotrichum state of this fungus was also compared with C. lindemuthianum and C. musae in terms of morphology of conidia and setae and growth on PDA etc. This fungus was identified as Glomerella cingulata(Stonem.) Spaulding & Schrenk(Colletotrichum gloeosporioides(Penz. Sacc.).

Effects of Streptomyces griseofuscus 200401 on Growth of Pepper Plants and Phytophthora Blight by Phytophthora capsici (토양 분리 방선균 Streptomyces griseofuscus 200401의 고추 생육 촉진과 역병 발생 억제 효과)

  • Lim Tae-Heon;Kwon Soon-Youl;Kim Jin-Ho
    • Research in Plant Disease
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    • v.12 no.1
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    • pp.46-50
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    • 2006
  • The microorganisms with the antifungal activity against Phytophthora capsici and Colletotrichum acutatum and the plant growth-promoting activity were screened from a forest and natural fields of Gajang-Dong, Sangju-city. One of the isolates, Streptomyces griseofuscus 200401, was selected as a good plant growth-promoting strain in this study. In greenhouse test, the number of leaf, fresh weight, and dry weight of pepper plants, that were grown with treatment of culture suspension or powder containing S. griseofuscus 200401, were higher than those without the bacterial cells. Cultivation of S. griseofuscus 200401 strain for 7 days in a nutrient rich medium produced ammonium chloride up to 0.13 ${\mu}g/ml$ in the culture solution of S. griseofuscus. Treatment of the selected strain significantly reduced the severity of the late blight of pepper plants to show the equivalent disease control activity to chemical fungicide. This study suggests that S. griseofuscus 200401 strain could be a potential biological agent with the biocontrol activity and the plant growth-promoting activity.

Effect of Sodium Hypochlorite Treatment on Incidence of Seed-borne Fungi in Several Crop Seeds (Sodium Hypochlorite 처리가 몇가지 작물의 종자소독에 미치는 효과)

  • Ku, Ja Hyeong;Yu, Seung Hun;Lee, Hyang Burm
    • Korean Journal of Agricultural Science
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    • v.20 no.1
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    • pp.18-24
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    • 1993
  • This experiment was focused on determining the potential of sodium hypochlorite (NaOCl) as fungicide against seed-borne fungi. Effects of NaOCl to control seed-borne fungi were compared to the those of Benlate T in several crop seeds. 1. The effect of NaOCl on disinfection of sesame seeds without imparing germination was highest in the range of 1-2% solution for 10 min. Also, a 30 min immersion of rice seed in 1-2% solution reduced incedence of seed-borne fungi. 2. Alternaria spp. in seeds of radish and chinese cabbage and Colletoricum spp. in pepper were significantly reduced by a 10 min immersion of seeds in 1% NaOCl. 3. The effective control range of NaOCl for seed-borne fungi was much wider than that of Benlate T in sesame seeds. No clear difference between chemicals was found in rice seeds. However, germination of seeds were impaired at 1-2% NaOCl immersion for more than 1 hour.

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Enhancement of Antifungal Activity of Anthracnose in Pepper by Nanopaticles of Thiamine Di-lauryl Sulfate (비타민 B1 유도체(Thiamine Di-lauryl Sulfate:TDS)의 나노입자화를 통한 고추탄저병균의 항진균 활성 증진)

  • Seo, Yong-Chang;Cho, Jeong-Sub;Jeong, Hae-Yoon;Yim, Tae-Bin;Cho, Kyoung-Sook;Lee, Tae-Woo;Jeong, Myoung-Hoon;Lee, Gang-Hyeong;Kim, Sung-Il;Yoon, Won-Byung;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.3
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    • pp.198-204
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    • 2011
  • This study was performed to enhance antifungal activity of anthracnose in chili pepper by nanopaticles of thiamine di-lauryl sulfate (TDS) through high pressure homogenization process. Yield of TDS was 79.14% by reaction of thiamine hydrochloride and sodium lauryl sulfate. TDS nanopaticle solution was manufactured through high pressure homogenization process. The turbidity of nanoparticles solution was increased with increasing the concentration of TDS, and nanoparticles solution of 100 ppm was showed the highest turbidity with absorbance of 3.212. The size of nanoparticles solution was measured as average 258.6 nm by DLS. Nanoparticles solution of 100 ppm showed growth inhibition activity with higher than about 80% compared to the control group against Colletotrichum gloeosporioides. Finally, nanoparticles solution was increased effectively the penetration of the TDS nanopaticles on attached cell membrane of hyphae and started to destruct the cells under microscope observation. Consequently, we suggested that the TDS nanoparticle solution by high pressure homogenization process might be suitable biochemical pesticides for improving the antifungal activities against anthracnose in pepper.

Cultural characteristics of Bacillus velezensis HKB-1 in the water extract of the composted spent mushroom substrate of Lentinula edodes and biological control of Phytophthora blight disease of pepper (표고버섯 수확후배지 퇴비 추출물에서 Bacillus velezensis HKB-1의 배양적 특징 및 고추역병의 생물학적 방제)

  • Kim, Ja-Yoon;Seo, Hyun-Ji;Kang, Dae-Sun;Kang, Hee-Wan
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.272-278
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    • 2021
  • Bacillus velezensis HKB-1 was isolated from the composted spent mushroom substrate of Lentinula edodes (LeCSMS) and inhibited mycelial growth of phytopathogenic fungal species, Phythhopthora capsici, Collectotrichum coccodes and Fusarium oxysporium by more than 70%. B. velezensis HKB-1 showed bacterial growth rate 10 to 100 times higher than that of other commercial bacterial media in water extract of LeCSMS supplemented with 1% molasses. The LeCSMS medium was effective in promoting the growth of pepper and controlling Phytophthora blight disease of pepper.