• Title/Summary/Keyword: mycelial liquid culture

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Biocontrol Activity of Bacillus amyloliquefaciens CNU114001 against Fungal Plant Diseases

  • Ji, Seung Hyun;Paul, Narayan Chandra;Deng, Jian Xin;Kim, Young Sook;Yun, Bong-Sik;Yu, Seung Hun
    • Mycobiology
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    • v.41 no.4
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    • pp.234-242
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    • 2013
  • A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition.

The Cultural Characteristics of Paecilomyces tenuipes (눈꽃동충하초의 배양적 특성)

  • Kang, Bok-Hee;Lee, Sang-Han;Hur, Sang-Sun;Shin, Yong-Kyu;Lee, Dong-Sun;Chang, Hung-Bae;Song, Bong-Jun;Lee, Jin-Man
    • Food Science and Preservation
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    • v.17 no.3
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    • pp.365-369
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    • 2010
  • This study was conducted to investigate the cultural characteristics of Paecilomyces tenuipes PJ-1 separated from the nature. We investigated the effect of media, pH and temperature in growth of Paecilomyces tenuipes PJ-1 on solid culture media and the effect of inoculum size, glass beads in liquid culture condition. Paecilomyces tenuipes showed the most favorable growth on PDA among 5 different media. The optimum growth pH and temperature were at pH 6.0 and $25^{\circ}C$ on solid culture. Mycelial growth of P. tenuipes decreased rapidly above $35^{\circ}C$ and under $15^{\circ}C$. In liquid culture, the optimum inoculum size was 10.0% and the SED and PMV value were increased with adding glass bead and glass bead size in the range of glass bead 0~50 ea and size 3, 5 mm.

Optimization of Submerged Cultivation of Hericium erinaceum (Hericium erinaceum 액체배양의 최적화)

  • Jung, Jae-Hyun;Lee, Keun-Eok;Lee, Shin-Young
    • KSBB Journal
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    • v.21 no.2
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    • pp.96-102
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    • 2006
  • Recently, it has been known that Hericium erinaceum is a one of the very useful functional materials with great attention in mushroom processing industry. In present study, a liquid culture which was not studied systematically until now, was conducted as a method of cultivation for H. erinaceum, and also examined the characteristics of the liquid culture and conditions of process optimization. A good basal medium was selected through the cultivation of 16 species mushroom media and the optimum condition for medium and cultivation were chosen by response surface method. From these results, the optimum condition of medium for mushroom was 3% glucose, 0.2% yeast extract/peptone(1:1) and 0.1% $KH_2PO_4/MgSO_4$(1:1) and also the optimal culture condition was obtained at inoculum of 13.42%, temperature of $22.3^{\circ}C$ and pH of 5.7. The mycelial dry weight of 9 g/I was obtained under these conditions and this amount was about 1.7 times higher than that which were cultivated in basal medium for 8 days.

Selection of KYC 3270, a Cellulolytic Myxobacteria of Sorangium cellulosum, against Several Phytopathogens and a Potential Biocontrol Agent against Gray Mold in Stored Fruit

  • Kim, Sung-Taek;Yun, Sung-Chul
    • The Plant Pathology Journal
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    • v.27 no.3
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    • pp.257-265
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    • 2011
  • During 2002-2008 in Korea, 455 extracts from myxobacteria consisting of 318 cellulolytic and 137 bacteriolytic myxobacteria were isolated, which were then screened for antifungal activity against the phytopathogens Botrytis cinerea, Colletotrichum acutatum, Penicillium sp., Pyricularia grisea, and Phytophthora capsici. 204 isolates had antifungal activity, causing both a clear zone due to blocked spore germination and inhibition of mycelial growth; most (199) were from cellulolytic (Sorangium cellulosum) and only five were from bacteriolytic myxobacteria. B. cinerea, the best controlled among the five tested pathogens, had a unique group of antifungal isolates of myxobacterial extracts compared to the other pathogens' groups. Among seventy-nine bioactive myxobacteria, four isolates, KYC 3130, KYC 3247, KYC 3248 and KYC 3270, were selected and all were cellulolytic. Liquid culture filtrates of these four myxobacteria were applied to tomato, cherry tomato, strawberry, and kiwi fruits 5 h before inoculation with gray mold conidia; then the treated fruits were placed in an airtight container and the experiment was repeated six to eight times. Incidence (%) of gray mold on fruit of the infected control treatment was 84-98%, whereas it was only 5-21% after the KYC 3270 treatment. After KYC 3270 treatment of the four fruits, mold control was 79-95%, which was highest among the filtrates and statistically the same as treatment with fludioxonil, a registered chemical against gray mold of stored fruits.

Effect of Potassium Phosphonate on the Control of Phytophthora Root Rot of Lettuce in Hydroponics

  • Jee, Hyeong-Jin;Cho, Weon-Dae;Kim, Choong-Hoe
    • The Plant Pathology Journal
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    • v.18 no.3
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    • pp.142-146
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    • 2002
  • The effect of potassium phosphonate ($KH_2PO_3 or $K_2 HPO_3$) on the control of Phytophthora root rot of lettuce was evaluated in a liquid hydroponic culture. Phosphonate 100 ppm strongly inhibited mycelial growth of Phytophthora species in vitro but did not affect normal growth of lettuce in a greenhouse test. Application of the chemical before infection showed over 94% control value, while it was less than 35% when applied after infection. In a field trial, phosphonate 100 ppm, which was directly supplemented into the nutrient solution, satisfactorily controlled the disease as it did not develop until 28 days after transplanting and remained at less than 2% infection rate at the end of cultivation. Meanwhile, in the control plot, the disease initiated at 7 days after transplanting and developed rapidly reaching over 70% infection rate at 28 days. Population density of the causal pathogen, R drechsleri, in a heavily infested farm was 22.0-25.0 cfu/100 ml of nutrient solution. However, it decreased to 1.3-2.0 cfu/100 ml at 7 days after treatment with phosphonate 200 ppm.

Biocontrol with Myxococcus sp. KYC 1126 Against Anthracnose in Hot Pepper

  • Kim, Sung-Taek;Yun, Sung-Chul
    • The Plant Pathology Journal
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    • v.27 no.2
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    • pp.156-163
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    • 2011
  • Antifungal activity of three Myxococcus spp., KYC 1126, 1136, and 2001, was tested in vitro against three phytopathogenic fungi (Botrytis cinerea, Colletotrichum acutatum, and Pyricularia grisea). Spore germination and mycelial growth of the three pathogenic fungi were completely inhibited by bioactive substances from a myxobacterium KYC 1126. In addition, the activity of KYC 1126 was fungicidal, but liquid culture filtrate of KYC 1126 did not affect protoplast reversion in C. acutatum. A bioassay of KYC 1126 filtrate against anthracnose in hot pepper was conducted in the greenhouse and field at 2009 and 2010. The incidence of anthracnose in control seedlings was 74%, but was reduced to 29% after KYC 1126 treatment. The control value with KYC 1126 was 60% while that with the fungicide dithianon was 42%. In the greenhouse, disease incidence with KYC 1126 was consistentely 10-35% lower than with fungicide as a positive control. The control value with KYC 1126 was 13.4% and 41.0%, whereas that with the fungicide was 52.3% and 63% in 2009 and 2010, respectively. Although anti-anthracnose activity of KYC 1126 was not maintained for long time in the field, the bacteriolytic myxobacterium KYC 1126 could be a prospective biocontrol agent.

Selection and a 3-Year Field Trial of Sorangium cellulosum KYC 3262 Against Anthracnose in Hot Pepper

  • Yun, Sung-Chul
    • The Plant Pathology Journal
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    • v.30 no.3
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    • pp.279-287
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    • 2014
  • KYC 3262 was selected as a biocontrol agent against anthracnose on hot pepper from 813 extracts of myxobacterial isolates. Dual culture with Colletotrichum acutatum and 813 myxobacterial extracts was conducted, and 19 extracts were selected that inhibited germination and mycelial growth of C. acutatum. All selections were Sorangium cellulosum, which are cellulolytic myxobacteria from soil. With the infection bioassay on detached fruits in airtight containers, KYC 3262, KYC 3512, KYC 3279, and KYC 3584 were selected. The listed four myxobacteria were cultured in CSG/1 liquid media, and harvested filtrates were sprayed on the infected fruits. KYC 3262 was selected from the studies of attached fruit in a greenhouse study. KYC 3262 filtrate was applied for 3 years (from 2011 to 2013) in a field study in Asan, Republic of Korea. Control values of the KYC 3262 in the field were 31%, 89%, and 82% in 2011, 2012, and 2013, whereas values of the fungicide spray treatment were 19%, 97%, and 91%, respectively. Yields (kg/20 plants) of the KYC 3262 were 2.66 kg and 18.6 kg in 2011 and 2013, respectively, and those of the fungicide treatment were 2.0 kg and 20.2 kg, in 2011 and 2013, respectively.

Isolation of Antimicrobial Substances from Hericium erinaceum

  • Kim, Dong-Myong;Pyun, Chul-Woo;Ko, Han-Gyu;Park, Won-Mok
    • Mycobiology
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    • v.28 no.1
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    • pp.33-38
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    • 2000
  • Mycelium of Hericium erinaceum isolate KU-1 was cultured in liquid medium (HL medium) and solid medium (Ko medium) at pH 4.0 in $28^{\circ}C$. 1.0% glucose or fructose was the most favorable carbon source, and 0.2% amonium acetate or $NaNO_3$ was an exellent nitrogen source for mycelial growth as well as production of antimicrobial substances. The mixture of saw dust 70% with rice bran 30% (SR medium) was the substrate for formation of sporophores. The active substrates in extracts from mycelium, culture filtrate and fruiting body were separated by TLC. The solvent for TLC was EtOAc: Chloroform: MeOH (10 : 5 : 10). Phenol-like substances appeared at Rf $0.5{\sim}0.9$, and fatty acid-like substances appeared at Rf $0.1{\sim}0.2$. The purified materials from the extracts showed antimicrobial effects to Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Aspergillus niger, Candida albicans and Microsporum gypseum. The S. aureus was the most inhibited. Minimal inhibitory concentration (MIC) of purified white powder and the Hercenone derivatives against S. aureus were $5.65\;{\mu}g/ml$ and $1.85\;{\mu}g/ml$, respectively.

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Establishment of artificial cultivation technique of Cordyceps militaris using a grain media (곡물배지를 이용한 번데기동충하초 재배기술)

  • Yu, Young-Jin;Seo, Kyoung-Won;Lee, Kong-Joon;Choi, Dong-Chil;Choi, Joung-Sik;Park, Joung-Sik;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.5 no.2
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    • pp.81-85
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    • 2007
  • This study was carried out to investigate the conditions of liquid culture and fruiting body formation of Cordyceps militaris using grain mediacontained the brown rice and black purple rice. Liquid mycelial growth and density of Cordyceps militaris were the highest in the medium of malt extract 0.7% and pH 5.0~5.5. The optimal condition of formation the fruiting bodies were shown to be temperature of $17^{\circ}C$, light intensity of 600 lux, $C0_2$ of 500ppm and humidity intensity of 95% produced highest amount of fruit bodies.

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Antimicrobial Activity of the Coriolus versicolor Liquid Culture Extracts Against Antibiotic Resistant Bacteria and Purification of Active Substance (구름버섯 균사체 배양 추출물의 복합내성 세균에 대한 항균활성 및 활성물질의 정제)

  • Lee, Jung-Sun;Kim, Taeg;Lee, Yoon-Hi;Jin, Cheng-Min;Kim, Hyun-Guell;Kim, Woo-Jung;Oh, Duek-Chul;Park, Yong-Il
    • The Korean Journal of Mycology
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    • v.34 no.2
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    • pp.92-97
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    • 2006
  • The liquid culture extract of Coriolus versicolor was prepared by directly boiling the whole culture broth 7 days after incubation in 12% citrus extract medium. After removal of mycelial debris through filtration, this extract was further extracted with equal volume of ethyl acetate (1 : 1, v/v). The ethyl acetate extracts showed significant antibacterial activities against Stapylococcus aureus CCARM3230 and Psudomonas aeruginosa CCARM2171, which are resistant to several antibiotics. The most active fraction was eluted from a silica gel column with a mixture of dichloromethane and methanol (9 : 1, v/v) and the purity of this active substance was confirmed by HPLC analysis. The results suggest that the purified active substance could be a good source for the development of a new antimicrobial agent, especially for the treatment of antibiotic resistant bacteria.