• Title/Summary/Keyword: mycelial cultivation

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Kinetics of Cell Growth and Cyclosporin A Production by Tolypocladium inflatum when Scaling Up from Shake Flask to Bioreactor

  • El Enshasy, H.;Fattah, Y. Abdel;Atta, A.;Anwar, M.;Omar, H.;Magd, S. Abou El;Zahra, R. Abou
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.128-134
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    • 2008
  • The kinetics of cell growth and Cyclosporin A (Cyc A) production by Tolypocladium inflatum were studied in shake flasks and bioreactors under controlled and uncontrolled pH conditions. In the case of the shake flask, the production time was extended to 226 h and the maximal antibiotic concentration was 76 mg/l. When scaling up the cultivation process to a bioreactor level, the production time was reduced to only 70h with a significant increase in both the cell growth and the antibiotic production. The maximal dry cell weights in the case of the controlled pH and uncontrolled pH cultures in the bioreactor were 22.4g/l and 14.2g/l, respectively. The corresponding maximal dry cell weight values did not exceed 7.25g/l with the shake flask cultures. The maximal values for Cyc A production were 144.72 and 131.4 mg/l for the controlled and uncontrolled pH cultures, respectively. It is also worth noting that a significant reduction was observed in both the dry cell mass and the antibiotic concentration after the Cyc A production phase, whereas the highest rate of antibiotic degradation was observed in the stirred tank bioreactor with an uncontrolled pH. Morphological characterization of the micromorphological cell growth (mycelial/pellet forms) was also performed during cultivation in the bioreactor.

Secretion of the iron containing superoxide dismutase of Streptomyces subrutilus P5 (Streptomyces subrutilus P5가 생산하는 철 함유 superoxide dismutase의 분비)

  • Park, Jae-seung;Kim, Jae-heon
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.108-114
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    • 2015
  • We tried to analyze the growth time for secretion of the iron containing superoxide dismutase by comparing the intra-and extracellular enzyme activity from Streptomyces subrutilus P5 and analyze possible genetic information for this enzyme secretion. The mycelial dry weights and glucose concentrations in culture filtrates were determined during growth. Glucose was consumed rapidly during logarithmic growth phase and almost exhausted at 24 h of cultivation. While the intracellular activity of iron containing superoxide dismutase was first appeared at three hours, the extracellular activity of this enzyme appeared from 7.5 h of cultivation, early logarithmic growth phase. This early presence of the superoxide dismutase might not be the result of cell lysis but active secretion pathway. There was no information for signal peptide responsible for the enzyme secretion in sodF. However, we found a type three secretion box in the promoter region of sodF that has been known for the genes of type III secreted proteins in other bacteria. This is the first report on the possible existence of type III secretion in Streptomyces.

Growth characteristics of oyster mushroom in bottle cultivation with addition of cottonseed meal (느타리버섯 재배에서 면실박의 첨가비율에 따른 생육 특성)

  • Lee, Chan-Jung;Lee, Eun-Ji;Park, Hye-Sung;Kong, Won-Sik
    • Journal of Mushroom
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    • v.17 no.3
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    • pp.162-166
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    • 2019
  • This study was carried out to investigate the optimum level of cottonseed meal in bottle cultivation of oyster mushrooms. Mycelial growth was slightly slower than that of the control at a cottonseed meal level ${\leq}10%$. Pileus diameter and thickness were highest at 14%, and stipe diameter was highest at 12% cotton seed meal. The hardness of pileus and stipe was greatest at 16% cotton seed meal. L-values tended to be lower at low levels of cotton seed meal, but the a- and b-values showed no significant difference among the treatments. The yield of fruiting body was 147.2 g/850 ml in the control and slightly higher at 147.6 g/850 ml using 12% cotton seed meal.

Control Efficacy of Bacillus velezensis AFB2-2 against Potato Late Blight Caused by Phytophthora infestans in Organic Potato Cultivation

  • Kim, Min Jeong;Shim, Chang Ki;Park, Jong-Ho
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.580-595
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    • 2021
  • Although late blight is an important disease in ecofriendly potato cultivation in Korea, it is highly dependent on the use of eco-friendly agricultural materials and the development of biological control technology is low. It is a necessary to develop an effective biocontrol agent to inactivate late blight in the field. AFB2-2 strain is a gram-positive with peritrichous flagella. It can utilize 20 types of carbon sources, like L-arabinose, and D-trehalose at 35℃. The optimal growth temperature of the strain is 37℃. It can survive at 20-50℃ in tryptic soy broth. The maximum salt concentration tolerated by AFB2-2 strain is 7.5% NaCl. AFB2-2 strain inhibited the mycelial growth of seven plant pathogens by an average inhibitory zone of 10.2 mm or more. Among the concentrations of AFB2-2, 107 cfu/ml showed the highest control value of 85.7% in the greenhouse. Among the three concentrations of AFB2-2, the disease incidence and severity of potato late blight at 107 cfu/ml was lowest at 0.07 and 6.7, respectively. The nucleotide sequences of AFB2-2 strain were searched in the NCBI GenBank; Bacillus siamensis strain KCTC 13613, Bacillus velezensis strain CR-502, and Bacillus amyloliquefaciens strain DSM7 were found to have a genetic similarity of 99.7%, 99.7%, and 99.5%, respectively. The AFB2-2 strain was found to harbor the biosynthetic genes for bacillomycin D, iturin, and surfactin. Obtained data recommended that the B. velezensis AFB2-2 strain could be considered as a promising biocontrol agent for P. infestans in the field.

Development of medium for Pleurotus eryngii cultivation using Kojongsi persimmon peels (감과피를 이용한 큰느타리버섯 배지 개발)

  • Kim, Chul Hwan;Kim, Hye Soo;Kim, Hong Chul;Kwon, Hyun Sook;Cheong, Jong-Chun;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.314-318
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    • 2015
  • This study was carried out to investigated the avaiability of dried Koojongsi persimmon peels (KPP) as a useful mushroom medium using Pleurotus eryngii ASI 2312. Mushroom cultivation medium used in this study was mixed with medium mixture, corn cob and sawdust (220:65:15, v/v). Dried KPP was replaced mushroom cultivation medium (control) with 5, 10, 15, 20, 30, 40, 50% dried KPP. The T-N content of dried KPP treatments decreased to increase replaced ratio of the dried KPP and C/N ratio was increased to increase replaced ratio of the dried KPP. But T-C content of dried KPP treatments was similar to untreated control. The average cultivating periods of mycelium on dried KPP treatments was delayed to increase replaced ratio of the dried KPP and cultivating periods was delayed over 30% dried KPP treatments. The length of stipe of dried KPP treatments was longer than that of the untreated control to increase replaced ratio of the dried KPP and thickness of stipes was tend to be thinner than that of the untreated control to increase replaced ratio of the dried KPP. The moisture, carbohydrate, crude protein and crude ash content of mycelial were similar to untreated control, but crude fatty acid was increased to increase replaced ratio of the dried KPP. The ${\beta}-glucan$ content of 10% and 15% treatments were higher than untreatment control. The results based on cultivation yield and ${\beta}-glucan$ content indicated that optimal mixture ratio dried KPP was 15%.

Effects of Using Cold Water on Mixing Sawdust Media for Flammulina velutipes (고온기 팽이버섯 병재배 배지제조시 저온수 이용 효과)

  • Cheong, Jong-Chun;Jhune, Chang-Sung;Kim, Seung-Hwan;Won, Hang-Yeon;Kwon, Jae-Geon
    • Journal of Mushroom
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    • v.3 no.4
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    • pp.140-144
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    • 2005
  • This study was conducted to verify the cause of suppression symptom in mycelial growth during summer and to be able to establish a countermeasure. Cultivation of Flammulina velutipes was experimented with varying elapsed time of 0, 3, 6, 9 hours after mixing the sawdust media and two kinds of water temperature (normal water, $24^{\circ}C$ and cold water, $6^{\circ}C$) for mixing sawdust media. There were trends of increased media temperature from $24^{\circ}C$ to $31^{\circ}C$ and decreased pH from 6.5 to 5.2~5.6 with varying elapsed time from mixing the media to sterilization. Bacterial density also increased with bacterial density in Medium $24^{\circ}C$ being 1.9~4.1 times higher than that in Medium 6. Growth of F. velutipes was delayed with dual culture of bacteria isolated from sawdust media. Total nitrogen content of sawdust media was lowered by elapsed time from mixing the media to sterilization. The use of normal water($24^{\circ}C$) delayed mushroom growth by 1~2 days compared with that of cold water($6^{\circ}C$). Furthermore, mycelial growth of F. velutipes in the bottle cultivation ceased 9 hours after mixing the media. Primordia formation of F. velutipes was delayed by 1~3 days by elapsed time after mixing sawdust media, while fruit-body yield decreased by 7~12% 6 hours after mixing the media. Fruit-body yield was more stabilized with the use of cold water($6^{\circ}C$) than with that of normal water($24^{\circ}C$). Results showed that it is effective to use cold water as low as $6^{\circ}C$ in mixing media for F. velutipes cultivation, especially during summer when environmental temperature is high, high pressure sterilization after bottling work can prevent bacterial propagation in the media and can stabilize media ingredient.

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Characteristics of the gene resources and selected strains of Agrocybe cylindracea (버들송이버섯(Agrocybe cylindracea)유전자원 및 선발계통의 특성)

  • Heo, Byong-Soo;Yoo, Young-Jin;Seo, Sang-Young;Choi, Kyu-Hwan;Choi, Young Min;Kwon, Seog-Ju;Jang, Kab-Yeul
    • Journal of Mushroom
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    • v.17 no.2
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    • pp.52-63
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    • 2019
  • Agrocybe spp. belongs to the Agaricales order, Bolbitiaceae family, and Agrocybe genus. In Korea, so far, it has been cultivated through bottle cultivation; therefore, this study was conducted for the development of a new cultivar using the bag cultivation method for quantitative improvement. Thirty-three gene resources of Agrocybe spp. were collected and their quantity and characteristics of bag cultivation were examined. Next, 5 kinds of crossing parents were selected based on the cultivation period and shape of the fruit body. Seven strains were selected through 3 cross combinations. The 7 selected strains and the comparative cultivar 'Cham' were cultivated in a bag. As a result, the cultivation period was 49 days for 'JBAC15-1' and 50 days for 'JBAC15-6' which are 4 days and 3 days less than the cultivation period of the comparative cultivar 'Cham'(53 days), respectively. Cultivation periods of other strains except for 'JBAC15-1' and 'JBAC15-1' were longer than that of the comparative cultivar'Cham'. The best ratio of primordia formation among the selected strains was found to be that of 'JBAC15-1' with 96.1% followed by 'JBAC15-6' with 94.5%. These rates were 3.1% and 1.5% higher than the ratio of primordia formation of the comparative cultivar 'Cham', which is found to be 93.0%. The quantity was maximum in the 'JBAC15-1' cultivar with 176.8 g per bag followed by 'JBAC15-6' with 168.7 g per bag. The quantities were 10% and 5% more than the comparative cultivar 'Cham' with 160.7 g per bag. Based on these results, 'JBAC15-1' and 'JBAC15-6', which had shorter cultivation periods and more quantities than the comparative cultivar 'Cham' were finally selected. For the selected strains of 'JBAC15-1' and 'JBAC15-6', mycelial growth was observed to be optimal on PDA medium and the optimum temperature was $27.5^{\circ}C$. The optimum pH was pH 5 for 'JBAC15-1' and pH 6 for 'JBAC15-6'. The color of the pileus of the fruit body was dark brown in 'JBAC15-1' and 'Cham' and light brown in 'JBAC15-6'. The pileus was hemispherical in shape in both 'JBAC15-1' and 'Cham'. However, the colors of the stem were different - light brown in 'JBAC15-1', white in 'JBAC15-6', and ivory in the comparative cultivar 'Cham'.

Inhibition Effects of Silver Nanoparticles against Powdery Mildews on Cucumber and Pumpkin

  • Lamsal, Kabir;Kim, Sang-Woo;Jung, Jin-Hee;Kim, Yun-Seok;Kim, Kyoung-Su;Lee, Youn-Su
    • Mycobiology
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    • v.39 no.1
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    • pp.26-32
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    • 2011
  • Powdery mildew is one of the most devastating diseases in cucurbits. Crop yield can decline as the disease severity increases. In this study, we evaluated the effect of silver nanoparticles against powdery mildew under different cultivation conditions in vitro and in vivo. Silver nanoparticles (WA-CV-WA13B) at various concentrations were applied before and after disease outbreak in plants to determine antifungal activities. In the field tests, the application of 100 ppm silver nanoparticles showed the highest inhibition rate for both before and after the outbreak of disease on cucumbers and pumpkins. Also, the application of 100 ppm silver nanoparticles showed maximum inhibition for the growth of fungal hyphae and conidial germination in in vivo tests. Scanning electron microscope results indicated that the silver nanoparticles caused detrimental effects on both mycelial growth and conidial germination.

Root Rot of Bottle Gourd Stock of Watermelon Caused by Monosporascus cannonballus in Korea (수박 대목용 참박에 발생한 Monosporascus cannonballus에 의한 검은점뿌리썩음병(黑点根腐病))

  • 박경석;남상현;김충회
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.175-180
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    • 1994
  • The fungal pathogen Monosporascus annonballus was first isolated in Korea from the rotted roots of bottle gourd stocks of collapsed watermelon plants in fields near Chochiwon, Choongnam province in July, 1993. Perithecia of M. cannonballus were dark brown to black, globose, 220~570 ${\mu}{\textrm}{m}$ in diam. and had many asci. Asci are hyaline, clavate to pyriform, and 50~120$\times$35~570 ${\mu}{\textrm}{m}$ in size. Ascospores were aseptate, dark brown to black, globose, 25~45$\times$30~50 ${\mu}{\textrm}{m}$ in diam, and borne singly in each ascus. The fungus grew in the temperature range of 4 to 34$^{\circ}C$, best at 3$0^{\circ}C$. The optimum pH for growth was 6.8. Mycelial growth rate of M. cannonballus was 25.5mm/day on PDA at 26$^{\circ}C$. Perithecia began to form after 20-day-growth on PDA and produced mature asci after 30 days or later. In the greenhouse inoculation tests, the fungus developed water-soaked lesions on roots of bottle gourd seedlings and was then reisolated from the lesions. Severed damages on watermelon plants by M. cannonballus are greatly concerned in Korea, since no stocks used for watermelon cultivation have reported to be resistant to the fungus.

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Screening of Botanicals Against Root Knot Disease Complex in Mulberry (Morus indica L.)

  • Naik V. Nishitha;Sharma D.D.;Govindaiah Govindaiah;Chowdary N.B.;Mala V.R.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.57-61
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    • 2007
  • For developing an integrated eco-friendly package against root knot disease complex of mulberry caused by the association of Meloidogyne incognita with Fusarium solani and F. oxysporum causing serious loss in terms of leaf yield and quality during cultivation, twenty botanical extracts at 5, 10 & 20% concentrations were screened under in vitro conditions. Among the extracts, Allium sativum followed by Lasownia inermis were found to be effective at 20% concentration against both the virulent fungi and nematode. Both the extracts reduced the mycelial growth of virulent fungi to an extent of 76-100%, inhibited the hatching of nematode eggs by 80-90% and 76-85% larval mortality over the control. The other extracts were found either moderately or poorly effective in reducing the growth of fungi, hatching of nematode eggs and enhancing the mortality of larvae. The two effective botanical extracts, which rated as strong inhibitors against both nematode and virulent fungi, can be utilized in developing an integrated ecofriendly technology for better management of root knot disease complex in mulberry.