• Title/Summary/Keyword: Broth media

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Development of an Efficient Method of Screening for Watermelon Plants Resistant to Fusarium oxysporum f. sp. niveum (수박 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Jo, Eun Ju;Lee, Ji Hyun;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.409-419
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    • 2015
  • This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor $1{\alpha}$) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of $1.0{\times}10^5-1.0{\times}10^6conidia{\cdot}mL^{-1}$ for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at $25^{\circ}C$ for 3 weeks.

Phytotoxicity of Endophytic Fungi and Characterization of a Phytotoxin Isolated from Gliocladium catenulatum from Pinus densiflora (식물내생곰팡이의 제초활성 검정 및 소나무에서 분리한 Gliocladium catenulatum이 생산하는 제초활성 물질의 특성 규명)

  • Choi, Gyung-Ja;Park, Joong-Hyeop;Kim, Heung-Tae;Lee, Seon-Woo;Choi, Jung-Sup;Hong, Kyung-Sik;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.8-15
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    • 2004
  • This study was conducted to discover new phytotoxins which may be used as lead molecules for the development of new herbicides. A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 8 locations in Korea. Their herbicidal activities were screened in vivo by herbicidal and duckweed bioassays after they were cultured in potato dextrose broth and rice solid media. Both fermentation broth and solid culture extract of Gliocladium catenulatum F0006 isolated from red pine (Pinus densiflora) showed 70% herbicidal activity only against cocklebur (Xanthium strumarium) out of the 10 weeds tested. Solid culture extract of F0034 isolated from arrowroot (Pueraria thunbergiana) exhibited 20 to 100% herbicidal activities against all of the weeds. Especially, shattercane (Sorghum bicolor), barnyardgrass (Echinochloa crus-galli), large crabgrass (Digitaria sanguinalis), and fall pauicum (Panicum dichtomiflorum) were sensitive to the solid culture extract of F0034. In addition, solid culture extract of F0043 isolated from red pine displayed 20% to 70% herbicidal activities only against 5 grass species, but not against 5 broad-leaf plant species. On the other hand, as the results of duckweed assay, 8 fermentation broths showed 100% growth inhibitory activity at concentrations less than 5.0% of culture supernatants and 12 solid cultures had a potent inhibitory activity against duckweed growth. A toxic metabolite was purified from the solid cultures of G. catenulatum F0006 by repeated column chromatography and bioassay. It caused a phytotoxic syndrome only on cocklebur out of the 10 weeds tested; it completely killed cocklebur seedlings at $500\;{\mu}g/ml$ and showed 85% herbicidal activity against cocklebur at $100\;{\mu}g/ml$. The molecular weight of the toxic metabolite is 238 daltons and its structure determination is underway.

The Growth Characteristics of Pleurotus eryngii (큰느타리버섯의 자실체 생육특성)

  • Ryu, Jae-San;Kim, Min-Keun;Kwon, Jin-Hyeuk;Cho, Sook-Hyun;Kim, Nak-Ku;Rho, Chi-Wong;Lee, Chun-Hee;Ro, Hyeon-Su;Lee, Hyun-Sook
    • The Korean Journal of Mycology
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    • v.35 no.1
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    • pp.47-53
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    • 2007
  • In this study, we investigated the properties of incubation and growing of Pleurotus eryngii in addition to the mycological properties to use them as basic data for breeding. The speed of mycelial growth on the MCM was faster than on the PDA. The biomass in the PDB broth culture was higher than in the MCM and YMG broth culture. KNR2515 and KNR2516 required 19 days for growth of mycelia on commercial sawdust media. KNR2503 required 6.5 days and 15.3 days for pin-heading and harvesting, respectively. In morphological properties by the mushroom, the heights of KNR2312 and KNR2322 were 122.7 and 121.0 mm, respectively. The thickness of KNR2322 and KNR2513 were 39.8 mm and 31.3 mm, respectively. The weight of KNR2524's fruiting body was 36.3 g, which is good as wild strain. The quality of fruiting body of KNR2503 was 4.0 in comparison to the score 7 of commercially cultivated strains. KNR2512 had the darkest color of pileus with L value 43.6. The slow growing strains, KNR2511, KNR2513, and KNR2512 had the bright pileus with L value 80. In morphological characteristics, KNR2511, KNR2513, and KNR2515 had white lamellar and plane pileus. The three strains are supposed to be the same group and KNR2516 and KNR2518 appeared to be related to the group. The commercially cultivated strains had convex pileus, KNR2502, KNR2503, KNR2504, KNR2521, and KNR2525 had infundibuliform, and the other strains had plane pileus. Several strains were valuable for breeding, JNR2503 for growth rate, KNR2512 for pileus color, and KNR2312, KNR2322, KNR2503, and KNR2513 for the quality.

EFFECT OF UNREACTED RESIN MONOMES ON THE ATIVITY OF CARIOGENIC BACTERIA (미반응 레진단량체가 우식유발성 세균의 활성에 미치는 영향)

  • Park, Seung-Kyu;Kim, Hwa-Sook;You, So-Young;Han, Jin-Ju;Kook, Joong-Ki;Lee, Nan-Young;Lee, Sang-Ho;Lee, Chang-Seop
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.4
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    • pp.684-695
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    • 2003
  • The aim of this study was to investigate the effect of composite resin components on proliferation and glucan synthesis by cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus. Light curing pit and fissure sealant was chosen for evluation. Specimens were eluted in deionized water for 10 minutes, 1, 12, and 24 hours. Extracts of specimens were diluted into 1/2, 1/4, and 1/8 with addition of BHI broth and BHI-YS. Bacteria were cultured in media included eluted components, and measured optical density($A_{600}$). The following results were obtained 1. 1/4 concentration of elutes for 10 minutes significantly inhibited the proliferation of S. mutans, whereas 1/2, 1/8 concentration of elutes stimulated it. Also, exacts, especially 1/2, 1/4 concentration, for 1 hours stimulated it. But exacts for 12, 24 hours had not effects on the proliferation of S. mutans. 2. 1/4 concentration of elutes for 10 minutes inhibited growth of S. sobrinus, whereas extracts for 1, 12, 24 hours had not effects on the proliferation of S. sobrinuss. 3. Extracts from composite resin stimulated total growth of S. mutans more than growth control group, where as inhibited it of S. sobrinus. 4. Extracts from composite resin, especially 1/4 concentration of it for 10 minutes increased the formation of water insoluble glucan of S. mutans. But elutes for 1, 12, 24 hours, and 1/8 concentration of it for 10 minutes inhibited it. 5. Except 1/4 concentration of elutes for 10 minutes, extracts decreased the formation of water insoluble glucan of S. sobrinus. 6. Total amount of formated glucan was 3-fold higher in S. mutans than in S. sobrinus.

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Biological activities of Fusarium isolates from soil and plants (토양 및 식물체로부터 분리한 Fusarium속 균주들의 생물활성)

  • Park, Joong-Hyeop;Choi, Gyung-Ja;Kim, Heung-Tae;Hong, Kyung-Sik;Song, Cheol;Kim, Jin-Seog;Kim, Jeong-Gyu;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.19-26
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    • 2000
  • In order to select potent bioactive isolates, 70 Fusarium isolates obtained from soil and 21 plant species were screened by antifungal, insecticidal, herbicidal, and duckweed bioassays after culturing in potato dextrose broth and rice solid media. Eight (11.4%) of the 70 liquid broth cultures showed disease-controlling activities more than 80% against at least one of the 6 plant diseases tested. Fusarium sp. FO-68 isolate exhibited the most potent antifungal activity; it controlled rice blast, wheat leaf rust, and barley powdery mildew with control values more than 95%. Out of 70 solid cultures, 21 (30.0%) controlled at least one plant disease more than 80% and F. equiseti FO-68 isolate showed disease-controlling activities more than 95% against 3 plant diseases such as rice blast, tomato late blight, and wheat leaf rust. As for tile insecticidal activities, 2 liquid and 1 solid cultures showed potent insecticidal activities against pest insects more than 80%, Liquid cultures of F. oxysporum FO-61 and Fusarium sp. FO-80 isolates exhibited insecticidal activities more than 80% against green peach aphid and diamondback moth, respectively. The solid culture of Fusarium sp. FO-510 isolate had 80% insecticidal activity against green peach aphid. However, none of liquid and solid cultures of the 70 Fusarium isolates showed potent herbicidal activities against 10 upland weeds. As the results of duckweed assay, 3 liquid cultures showed 70% growth inhibitory activity at concentrations less than 1.25% of culture supernatants and 9 solid cultures had a potent inhibitory activity against duckweed growth. On the other hand, there was a significant correlation between antifungal activities and herbicidal activities against duckweed of both liquid and solid cultures of tile 70 Fusarium isolates.

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Establishment of Optimal Production Conditions of Transglucosidase Produced by Aspergillus niger (Aspergillus niger가 생산하는 transglucosidase의 최적 생산 조건 확립)

  • Lee, Jun-Yeob;Gang, Seongho;Kim, Jong-Sik;Chung, Chungwook
    • Journal of Life Science
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    • v.28 no.8
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    • pp.969-976
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    • 2018
  • In this study, transglucosidase (TG), an enzyme produced by Aspergillus niger, synthesized isomaltooligosaccharide from ${\alpha}-(1{\rightarrow}4)$ linked substrates. The highest TG-producing A. niger KCTC6913 was selected from six kinds of species, and optimized TG producing conditions were established. Five different carbon sources (potato starch, sweet potato starch, corn starch, wheat starch, and dextrin) and three different nitrogen sources (yeast extract, malt extract, and beef extract) were tested to establish the carbon and nitrogen sources favorable for TG production. Measurements of TG activity after an initial culture at pH 5.0 for 15 days revealed that potato starch and yeast extract, which are basic culture media, resulted in the highest TG activity. In addition, A. niger KCTC6913 increased TG production under aerobic conditions and a controlled carbon/nitrogen ratio. In conclusion, to evaluate TG activity in the established optimal medium, it is confirmed that the basal and potato dextrose broth medium were used as a control, and the highest TG production was measured, which was highlighted in the established optimal medium.

Leaf Blight of Perilla Caused by Alternaria alternata (Alternaria alternata에 의한 들깨 잎마름병)

  • 차외진;김철승;송주희;김현주;이영병;문병주
    • Journal of Life Science
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    • v.12 no.6
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    • pp.708-714
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    • 2002
  • A new leaf blight was found on the perilla leaves at the major perilla-cultivating areas such as Kangdong in Busan and Miryang in Kyungnam province. Symptoms of the disease initially appeared on the edge of perilla leaves showing black necrosis and drying, and the infected leaves were finally fell down. The SD1 isolate showing strong pathogenicity and forming abundant conidial spores on the diseased lesions was isolated. Among the tested media, mycelial growth was abundant on PDA (Potato Dextrose Agar) medium at $25^{\circ}C$ under dark condition, but conidial formation was greater on V8A (V-8 juice A8ar) medium than that on PDA medium. Optimal temperatures for mycelial growth and conidial formation on PDA medium were respectively $25^{\circ}C$ and 3$0^{\circ}C$. The rate of conidial germination and the elongation of germ tube were more effective in 10% tomato juice than those in PDB (Potato Dextrose Broth) and sterile water. In 10% tomato juice, the rate of conidial germination and the length of germ tube were 100% after incubation for 24k and 535.2${\mu}{\textrm}{m}$ after incubation for 36hr, respectively. According to the result of pathogenicity, it revealed that conidial suspention with 10% tomato juice was the most effective for pathogenicity test showing as 100% of disease incidence, and the symptoms caused by artificial inoculum were same as those of naturally infected perilla. In this study, the SD1 isolate according to the results of morphological characteristics, the incubation characteristics and pathogenicity was firstly identified A. alternata, and named as leaf blight of perilla.

Morphological Characteristics of Pseudosclerotia of Grifola umbellata in In Vitro

  • Choi, Kyung-Dal;Lee, Kyung-Tae;Hur, Hyun;Hong, In-Pyo;Shim, Jae-Ouk;Lee, Youn-Su;Lee, Tae-Soo;Lee, Sang-Sun;Lee, Min-Woong
    • Mycobiology
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    • v.32 no.1
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    • pp.1-5
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    • 2004
  • The present study was carried out to investigate morphological characteristics of pseudosclerotia of Grifola umbellata formed by artificial cultures. Isolate G. umbellata DUM GUS-01 was obtained from sclerotium cultivated in field. The fungal isolate was cultured on PDYM broth, PDYMA(potato dextrose yeast malt agar) and oak sawdust media at $20^{\circ}C$ under the dark condition. G. umbellata DUM GUS-01 showed a volumetric increment of fungal lumps rather than mycelial growth. Particularly, G. umbellata DUM GUS-01 produced a large amount of melanin pigments in all culture treatments. The color of the fungal mass has been changed into grey gradually, and then formed melanized rind-like structure on its superficial part. The fungal structures which were covered with melanized rind-like layer were named as pseudosclerotia of G. umbellata. The pseudosclerotia of G. umbellata DUM GUS-01 formed a new white mycelial mass, which was swollen out of the melanized rind structure for its volumetric increment. When the pseudosclerotia were sectioned, their structure was discriminated from two structures such as a melanized rind-like structure layer formed by aggregation of aged mycelia and a white mycelial mass with high density. As results of scanning electron microscopic examination, the pseudosclerotia of G. umbellata DUM GUS-01 which were formed in in vitro conditions were similar to the sclerotia of G. umbellata cultivated in natural conditions except for the crystals formed in medula layer of natural sclerotia. Although size, solidity of rind structure and mycelial compactness of pseudosclerotia were more poor than those of natural sclerotia, the morphological structure and growth pattern of pseudosclerotia were very similar to those of natural sclerotia. Therefore, it is probable to induce pseudosclerotia to sclerotia of G. umbellata in in vitro conditions. Consequently, it seems that the induced pseudosclerotia can be used as inoculum sources to substitute natural sclerotia in field cultivation.

Characterization of a Chitinase Gene Exhibiting Antifungal Activity from a Biocontrol Bacterium Bacillus licheniformis N1

  • Lee, Kwang-Youll;Heo, Kwang-Ryool;Choi, Ki-Hyuck;Kong, Hyun-Gi;Nam, Jae-Sung;Yi, Young-Byung;Park, Seung-Hwan;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.344-351
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    • 2009
  • A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

Antimicrobial Efficacy of Fermented Dark Vinegar from Unpolished Rice (현미 발효 흑초의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.97-104
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    • 2015
  • Vinegar is a widely used acidic seasoning and can be manufactured using various methods and bases, including cereals, wheat, and fruits. Most studies on vinegar have been conducted to evaluate its antioxidant activity. In the present study, fermented dark vinegar (FDV) produced from unpolished rice was examined for its antimicrobial activity, biochemical content, including the amounts of sugar, total soluble sugar, organic acid, and free amino acids, and pH and physiological activity. The antimicrobial efficiency of FDV was assessed using the paper disc-agar diffusion method. FDV exhibited strong antimicrobial activity against the pathogenic bacteria and yeast strains that were tested. In fact, the activity of FDV was shown to be higher than that of the commercial antibiotics carbenicillin (50 µg/ml) and tetracycline (50 µg/ml) against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Yersinia enterocolitica, and Lodderomyces elongisporus. The antioxidant activity of FDV and ascorbic acid was evaluated. Using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, we found that FDV has the highest activity of the antioxidants. After spreading FDV onto tryptic soy broth and yeast extract-peptone-dextrose agar media, the microbial strains were isolated and characterized through physiological and biochemical analysis. Based on 16S ribosomal DNA sequence analysis, the isolated microorganisms exhibited a close similarity to Acetobacter papayae, Acetobacter pasteurianus, and Acetobacter peroxidans.