• Title/Summary/Keyword: Mycelial mass

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Importance of Strain Improvement and Control of Fungal cells Morphology for Enhanced Production of Protein-bound Polysaccharides(β-D-glucan) in Suspended Cultures of Phellinus linteus Mycelia (Phellinus linteus의 균사체 액상배양에서 단백다당체(β-D-glucan)의 생산성 향상을 위한 균주 개량과 배양형태 조절의 중요성)

  • Shin, Woo-Shik;Kwon, Yong Jung;Jeong, Yong-Seob;Chun, Gie-Taek
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.220-229
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    • 2009
  • Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

Biological control of Gray Mold Rot of Perilla Caused by Botrytis cinerea II. Formulation of Antagonistic Bacteria and Its Control Effect (들깨 잿빛곰팡이병의 생물학적 방제 II. 미생물농약의 제조 및 그 방제효과)

  • Moon, Byung-Ju;Kim, Choul-Soung;Song, Ju-Hee;Kim, Ju-Hee;Lee, Jae-Pil;Park, Hyean-Cheal;Shin, Dong-Bum
    • Research in Plant Disease
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    • v.8 no.3
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    • pp.184-188
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    • 2002
  • An antagonistic bacteria, Bacillus licheniformis Nl strain which effectively inhibited mycelial growth of gray mold rot pathogen, Botrytis cinerea was isolated from the rhizosphere of perilla crop. Powder soy formulation by B. lichentfomis Nl strain as a biocontrol agent was developed far the first time and estimated its control effect on perilla leaves in this study. First of all, far the mass production of antifungal metabolites of B. lichentfomis Nl strain in flask liquid culture, the most effective carbon and nitrogen source were selected as glucose and tryp-tone, respectively, For the formulation, vegetative biomass of B. licheniformis Nl strain from 5-day-old liquid culture in nutrient broth added glucose and tryptone was mixed with soy flour, rice flour glucose, FeSo$_4$~7$H_2O$, and MnCl$_2$. 4$H_2O$, and dried and pulverized. In plastic house test, powder soy formulation effectually controlled gray mold rot as the control value of 93.1 %, was more effective than chemical fungicide, benomyl showing the control value of 86.1%. Thus, development of powder soy formulation of B. lichentfomis Nl will aid large-scale application of biological control in field trials.

Development of artificial cultivation conditions on Tricholoma gigantium (왕송이버섯(Tricholoma gigantium)의 인공재배를 위한 환경조건 구명)

  • Jang, Kab-Yeul;Park, Jeong-Sik;Cheong, Jong-Chun;Kong, Won-Sik;Yoo, Young-Bok;Jhune, Chang-Sung;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.5 no.3_4
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    • pp.98-102
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    • 2007
  • Tricholoma gigantium, an edible mushroom, belongs to Tricholomataceae of Tricholoma and is distributed at Jeju-Do in Korea. It is also well-known as the medicinal mushroom in Taiwan. The cultivation method using the compost was developed in Korea in 1995. To develop a mass cultivation method and a superior strain, four strains were collected and tested. To establish the optimal cultivation conditions, various examinations were accomplished. Bag cultivation was more effective than box cultivation and the optimal relative humidity was more than 80%. Although the mycelial growth was tested in the substrate supplemented with different additives, such as rice bran and wheat bran, there's no significant difference between them. It suggested that the environmental conditions were more important than the substrate additives for cultivation.

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Characterization of Bacillus amyloliquefaciens DA12 Showing Potent Antifungal Activity against Mycotoxigenic Fusarium Species

  • Lee, Theresa;Park, Dami;Kim, Kihyun;Lim, Seong Mi;Yu, Nan Hee;Kim, Sosoo;Kim, Hwang-Yong;Jung, Kyu Seok;Jang, Ja Yeong;Park, Jong-Chul;Ham, Hyeonheui;Lee, Soohyung;Hong, Sung Kee;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.499-507
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    • 2017
  • In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of $31.3{\mu}g/ml$ and 100% at a concentration of $250{\mu}g/ml$. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.

Characteristics of Bed-log of Shiitake Damaged by Bjerkandera adusta and Antagonism between These Two Fungi (줄버섯 피해 표고골목의 특성 및 표고균과의 대치배양)

  • Bak, Won-Chull;Lee, Bong-Hun;Park, Young-Ae;Kim, Hyun-Seok
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.44-47
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    • 2011
  • A harmful fungus occurred seriously in bed-log of shiitake(Lentinula edodes) in Jangheung-Gun, Korea. The fungus was identified as Bjerkandera adusta by its morphology and ITS(Internal Transcribed Spacer) analysis. The fungus was reported as causal agent of stem-rot of Populus euramericana in Korea, but not reported in bed-log of shiitake until this notification. Thus, studies were made to investigate inside condition of bed-log of shiitake damaged by B. adusta, physiological characteristics of B. adusta and antagonism between these two fungi. First of all, B. adusta is white-rotting fungus like shiitake and wood-rotting condition is similar to that of shiitake. But, there are a lot of small spots in damaged wood tissue under bark which are not seen in case of shiitake. Optimal temperature for mycelial growth of B. adusta is ca. $30^{\circ}C$ while that of shiitake is ca. $25^{\circ}C$. When confrontation cultures were made between these two fungi under $15^{\circ}C$, $20^{\circ}C$, $25^{\circ}C$ and $30^{\circ}C$, B. adusta has antagonistic ability against shiitake in all the temperatures. From the results of experiments, if the bed-logs of shiitake are exposed to high temperature, there should be mass propagation of B. adusta, and shiitake mycelia will be seriously injured by the fungus. Therefore, to prevent the damage by B. adusta, it is needed to grow the mycelia of shiitake fast in the bed-log, and to avoid exposure of the bed-log to high temperature in summer.

Characterization of a new commercial strain 'Gumbit' in Pleurotus cornucopiae var. citrinopileatus (노랑느타리 품종 '금빛'의 특성)

  • Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Kim, In-Yeup;Oh, Se-Jong;Jhune, Chang-Sung
    • Journal of Mushroom
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    • v.4 no.3
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    • pp.83-87
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    • 2006
  • A new commercial strain "Gumbit" of golden oyster mushroom was developed by hyphal anastomosis. It was improved with hybridization between monokaryotic strain derived from ASI 2295 and ASI 2703. The optimum temperature of mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $19{\sim}24^{\circ}C$, respectively. The pileus was golden to brilliant yellow color. Commercial strain "Gumbit" was not as prolific as the more commonly cultivated Pleurotus ostreatus in the conversion of substrate mass to mushrooms. However, cultivator can save money for mushroom growing on high-temperature season as like summer in Korea. Since picking individual mushrooms is tedious and often damages the fragile fruiting bodies compared with other species of oyster mushrooms.

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Pink Mold Rot on Apple (Malus pumila var. dulcissima Koidz.) Caused by Trichothecium roseum (Pers.) Link ex Gray in Korea (Trichothecium roseum에 의한 사과 분홍빛열매썩음병 발생)

  • Kwon, Jin-Hyeuk;Kim, Min-Jeong;Shim, Chang-Ki;Jee, Hyeong-Jin;Lee, Sang-Dae
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.429-433
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    • 2014
  • In 2012, a pink mold rot was observed on apple (Malus pumila var. dulcissima Koidz.) at the Wholesale Market for Agricultural Products, Jinju, Korea. The first symptom of pink mold rot on apple fruit is a water-soaked appearance of the affected tissue and surface fruit formed pink mold rot, then became brown and produced a mass of powdery pink conidia spores. Colony was fast growing colonies, pinksh, zonate in diurnal rhythm, powdery from conidia. Optimum temperature for mycelial growth was $25^{\circ}C$. Conidia were hyaline, smooth, two-celled, thick-walled conidia with truncate bases, ellipsoidal to pyriform, and characteristically held together zig-zag chains and $12-26{\times}8-12{\mu}m$ in size. Conidiophore was erect, colorless, unbranched, and $4-5{\mu}m$ wide. On the basis of mycological characteristics, pathogenicity test, and molecular identification with the ITS region, the causal fungus was identified as Trichothecium roseum (Pers.) Link ex Gray.

Environmental Factors on the Development of Root Rot on Ginseng Caused by Cylindrocarpon destructans (인삼 뿌리썩음병 발병에 미치는 환경 요인)

  • Lee, Jung Sup;Han, Kyung Sook;Lee, Seong Chan;Soh, Jae Woo;Kim, Doo Wook
    • Research in Plant Disease
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    • v.20 no.2
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    • pp.87-94
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    • 2014
  • The fungus Cylindrocarpon destructans is the cause of root rot in many ginseng production areas in Korea. A total of 57 isolates of C. destructans were recovered from diseased roots in a survey of ginseng-growing fields from 2011-2012. Among these isolates, 37% were classified as highly virulent (causing lesions on unwounded mature roots) and 61% were weakly virulent(causing lesions only on previously wounded roots). Radial growth of highly and weakly virulent isolates on potato dextrose agar was highest at $20^{\circ}C$ and there was no growth at $35^{\circ}C$. Mycelial mass production was significantly (P = 0.05) lower at pH 7.0 compared with pH 5.0. To study the effects of pH (5.0 and 7.0) and wounding on disease development, ginseng roots were grown hydroponically in nutrient solution. Lesions were significantly larger (P < 0.01) at pH 5.0 compared with pH 7.0 and wounding enhanced disease by a highly virulent isolate at both pHs. In artificially infested soil, 2-yearold ginseng roots were most susceptible to Cylindrocarpon root rot among all root ages tested (1 to 4 years) when evaluated using a combined scale of disease incidence and severity. Root rot severity was significantly (P<0.05) enhanced by increasing the inoculum density from $3.5{\times}10^2cfu/g$ of soil to $2.0{\times}10^3cfu/g$ of soil.

Soft Rot on Citrus unshiu Caused by Rhizopus oryzae in Korea (Rhizopus oryzae에 의한 감귤 무름병)

  • Kwon, Jin-Hyeuk;Kim, Jin-Woo;Hyun, Jae-Wook;Lee, Yong-Hwan;Shim, Hong-Sik
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.78-81
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    • 2011
  • Soft rot caused by Rhizopus oryzae occurred on unshiu orange (Citrus unshiu Marc.) sampled from commercial markets in Jinju, Korea, 2010. The first symptom of soft rot on orange is a water-soaked appearance of the affected tissue. The infected parts later disintegrated into a mushy mass of disorganized cells followed by rapid softening of the diseased tissue. The lesion on orange was rapidly softened and rotted, then became brown or dark brown. Optimum temperature for mycelial growth of the causal fungus on potato dextrose agar was $30^{\circ}C$ and growth was still apparent at $37^{\circ}C$. Sporangiophores were $6{\sim}20\;{\mu}m$ in diameter. Sporangia were globose and $40{\sim}200\;{\mu}m$ in size. The color of sporangia was brownish-grey to blackish-grey at maturity. Sporangiospores were sub-globose, brownish- black streaked and $4{\sim}10\;{\mu}m$ in size. Columella were globose to sub-globose and $85{\sim}120\;{\mu}m$ in size. On the basis of mycological characteristics, pathogenicity test, and the ITS sequence analysis, the causal fungus was identified as Rhizopus oryzae. To our knowledge, this is the first report of soft rot caused by R. oryzae on unshiu orange in Korea.

Soft Rot on Peach Caused by Rhizopus oryzae in Korea (Rhizopus oryzae에 의한 복숭아 무름병)

  • Kwon, Jin-Hyeuk;Kang, Dong-Wan;Ha, Jeong-Seok;Kim, Jin-Woo;Kwak, Youn-Sig
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.65-68
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    • 2012
  • In July and August 2011, a disease suspected to be Rhizopus soft rot was observed on peach (Prunus persica var. vulgaris) at the Wholesale Market for Agricultural Products, Jinju, Korea. The first symptom of soft rot on peach is a water-soaked appearance of the affected tissue. The infected parts later disintegrated into a mushy mass of disorganized cells followed by rapid softening of the diseased tissue. The lesion on peach was rapidly softened and rotted, then became brown or dark brown. Optimum temperature for mycelial growth of the causal fungus on PDA was $30^{\circ}C$and growth was still apparent at $37^{\circ}C$Sporangiophores were 6~20 ${\mu}m$ in diameter. Sporangia were globose and 35~200 ${\mu}m$ in size. The color of sporangia was brownish-grey to blackish-grey at maturity. Sporangiospores were sub-globose, brownish- black streaked and 5~10 ${\mu}m$ in size. Columella were globose to sub-globose and 85~120 ${\mu}m$ in size. On the basis of mycological characteristics, pathogenicity test, and molecular identification, the causal fungus was identified as Rhizopus oryzae Went & Prinsen Geerligs. To our knowledge, this is the first report of soft rot caused by R. oryzae on peach in Korea.