• Title/Summary/Keyword: Liquid culture inoculum

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Selective colonization and removal of senescent flowers of zucchini squash by Trichoderma hrzianum YC459, a biocontrol agent for gray mold, Botrytis cinerea

  • Kim, Geun-Gon;Chung, Young-Ryun
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.90.2-91
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    • 2003
  • In commercial greenhouses, senescent flower petals or flowers of vegetables such as tomato, strawberry, hot pepper and zucchini squash were blighted to be removed from fruits within five days after spraying of Trichoderma harzianum YC459 (TORY), a biocontrol agent for the gray mold rot of vegetables caused by B. cinerea The mechanism for selective colonization of senescent floral tissues by T. harzianum YC459 was elucidated using fresh and senescent (Hays and 14days after flowering, respectively) floral tissues of zucchini squash (Cucurbita moschata Duchesne). The spores of T. hrzianum YC459 were produced more on agar and liquid culture media supplemented with 5% dry powder of senescent floral tissues than fresh tissues during 15days. Mycelial growth was also much better in the media with senescent tissues than with fresh tissues. Enzyme activities of amylase, polygalacturonase and cellulase in the liquid media which might be involved in the colonization of tissues by T. harzianum YC459 were compared. The activities of three enzymes were much higher in the media with senescent floral tissues than with fresh floral tissues reaching to the maximum during 9 to 12days of incubation. Based on the results, the removal of senescent floral tissues, a possible inoculum source of the pathogen, may be another mechanism for biocontrol of gray mold rot of vegetables by T. harzianum YC459.

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Production and Rooting of Shoots in Bioreactor Culture of Scrophularia buergeriana Miquel (생물반응기에서 현삼의 신초 형성과 발근)

  • Hahn, Suk-Hoon;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.2
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    • pp.117-122
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    • 2000
  • This study was carried out to know the factors affecting on shoot formation and rooting for stable and routine production of plantlets in bioreactor culture of Scrophularia buergeriana. Multiple shoots were formed effectively when explants were transplanted on the MS media with decreased concentration of $NH_4NO_3$ as 413mg/ l . Three hundred stem explants (0.8-1.0cm) was appeared as proper inoculation size in bioreactor culture. IBA (0.05mg/L) was more effective for rooting of the shoots in liquid as well as solid media. Six weeks long culture of explants in bioreactor gave better shoot shape for rooting on solid half-strength MS media.

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Favorable Condition for Mycelial Growth of Tricholoma matsutake (송이균 배양을 위한 균사생장 조건)

  • Kim, In-Yeup;Jung, Gwang-Reul;Han, Sang-Kuk;Cha, Joo-Young;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.33 no.1
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    • pp.22-29
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    • 2005
  • The main objectives of this research were to study the cultural and nutritional characteristics of Tricholoma matsutake and to establish its liquid culture system. The optimum growth of T. matsutake was observed in HA and TMM agar media. Similarly highest growth was observed in PDB and TMM liquid media. The optimal temperature for the mycelial growth was $25^{\circ}C$. The most suitable carbon source was dextrin among 12 different carbon sources tested. Yeast extract and peptone were best nitrogen sources among 17 different sources tested. The optimum mineral salts were $Fe_{2}(SO_{4})_{3}{\cdot}H_{2}O$ and KCl among 9 different sources tested. Shaking culture gave higher mycelial growth compared to stationary culture. Similarly, optimum medium amount for shaking culture was 100 ml per 250 ml flask. The highest mycelial growth was obtained when $5{\sim}7$ mycelial discs were inoculated in 100 ml of medium and incubated for $8{\sim}9$ weeks, respectively. The highest proportion of mycelial growth was observed at 40 : 1 ratio of medium to inoculum volume in 8 l air-lift fermenter.

Influence of Growth Conditions for the Production of Bacteriocin, Glycinecin, Produced by Xanthmonas campestris pv. glycines 8ra (콩 불마름병균의 생장 조건이 박테리오신인 glycinecin의 생성에 미치는 영향)

  • Woo Jung;Sunggi Heu;Cho, Yong-Sup
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.376-381
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    • 1998
  • Xanthomonas campestris pv. glycines 8ra causes bacterial pustule disease on susceptible soybean leaves and produces a bacteriocin, named glycinecin, against related bacteria such as Xanthomonas campestris pv. vesicatoria. The antimicrobial activity of the glycinecin was effective to most tested Xanthomonas species. X. c. pv. glycines 8ra was able to produce the glycinecin in liquid media as well as solid media. Maximal productivity of glycinecin was obtained at 3$0^{\circ}C$ in the early stationary phase of growth of the X. c. pv. glycines 8ra. The production of glycinecin was not dependent on the initial inoculum level but on cell density. Glycinecin was very sensitive to proteolytic enzymes such as trypsin and proteinase K but resistant to DNase and RNase. The culture supernatant of X. c. pv. glycines 8ra retained some of its antimicrobial activity after 15 min at 6$0^{\circ}C$. It is stable at wide range of pH. The glycinecin showed the bactericidal activity after the adsorption of the glycinecin to the sensitive bacterial cell.

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Condition of mycelial culture and inoculum volume of spawn on cultivation of Agrocybe cylindracea (버들송이의 균사배양조건 및 최적 접종량 설정)

  • Lee, Kee-Kwon;Yu, Young-Jin;Choi, Kyu-Hwan;Jeong, Jong-Seong
    • Journal of Mushroom
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    • v.10 no.1
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    • pp.21-28
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    • 2012
  • Studies were made to optimize the media composition and cultural condition for mycelial growth of Agrocybe cylindracea. Sawdust spawn of media composition for optimal growth was found to be pine sawdust combination of 30% wheat bran and poplar sawdust combination of 20% corn bran were the best of the optimal combination. The optimal concentration of white sugar was 1.0~1.5%. The nitrogen sources was found to be yeast extract and soybean powder. Also, optimal concentration were $0.7g/{\ell}$ and $0.1g/{\ell}$, respectively. The mineral sources of optimal medium compositions were $MgSO_4{\cdot}7H_2O\;0.3g/{\ell}$, $KH_2PO_4\;0.5g/{\ell}$ and $K_2HPO\;1.2g/{\ell}$. Optimal amount of inoculum for cultivation of A. cylindracea were $20{\sim}25g/850m{\ell}$ and $25m{\ell}/850m{\ell}$ in the sawdust spawn and liquid spawn, respectively.

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.

Large-scale Culture of Plant Cell and Tissue by Bioreactor System

  • Son, Sung-Ho;Park, Sung-Mee;Park, Seung -Yun;Kwon, Oh-Woung;Lee, Yun-Hee;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.1-7
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    • 1999
  • Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

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Investigation of the Condition of Fruiting Body Formation by Cordyceps scarabaeicola (풍뎅이동충하초(Cordyceps scarabaeicola)의 자실체 형성 조건)

  • Lee, Jae-Keun;Sung, Jae-Mo;Park, Young-Joon
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.11-17
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    • 2002
  • This experiment was carried out to study formation of fruitbody with Cordyceps scarabaeicola (EFCC C-252) isolate. This isolate was the one of best fruitbody formation on brown rice 60 g plus 30 g silkworm pupa media among EFCC C-251, EFCC C-252, EFCC C-1092 from EFCC (Entomopathogenic Fungal Culture Collection) of Kangwon National University. Fruiting body was formed only isolate EFCC C-252 among tested isolates on the medium of brown rice (60 g) and silkworm pupae (30 g). The optimal temperature and light for fruiting body formation were $25^{\circ}C$ and fluorescent light (300 lux). The maximal fruiting body formation was observed at 70 g of brown rice and 80 g of silkworm pupa medium which was treated separately. Fruiting body was formed maximally by 2 days interval of irrigation.

Competitive Growth and Attachment of Listeria monocytogenes and Lactococcus lactis ssp. lactis ATCC 11454

  • Lee, Shin-Ho;Frank, Joseph-F.
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.73-77
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    • 1992
  • The effect of a nisin-producing Lactococcus lactis spp. lactis (L. lactis) on the growth and attachment of Listeria monocytogenes Scott A and Brie 1 on stainless steel and their growth in Brain Heart Infusion broth was determined. Viable cells of Listeria decreased rapidly after 9~12 hr of incubation at $21^{\circ}C$ and after 6~9 hr of incubation at $32^{\circ}C$ in the presence of L. lactis. The number of L. monocytogenes Scott A attached to stainless steel in pure culture was $2.5{\times}10^3/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}2.3{\times}10^3/\textrm{cm}^2{\;}at{\;}32^{\circ}C$ after 48 hr of incubation, but was only $10/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}1.1{\times}10/\textrm{cm}^2{\;}at{\;}32^{\circ}C$ in the presence of L. lactis. Results from L. monocytogenes strain Brie 1 were similar to those from strain Scott A. The population of L. monocytogenes Scott A which attached to stainless steel with previously adherent L. lactis was $1.8{\times}10^2/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}8.2{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$, whereas the population attached to sterile stainless steel was $1.2{\times}10^3/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}2.1{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$. For L. monocytogenes Brie 1, the attached population of the control was $1.6{\times}10^4/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}3.2{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$, and on stainless steel with adherent L. lactis, it was $1.1{\times}10/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}6.9{\times}10/\textrm{cm}^2{\;}at{\;}32^{\circ}C$. Surface adherent L. lactis was less inhibitory to attachment of L. monocytogenes on stainless steel than a liquid culture inoculum. Listeria attached to stainless steel survived dry storage for 20 days both in the presence and absence of adherent lactococci.

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Effect of Medium Components and Culture Methods on Prothallus Propagation of Pteridium aquilinum var. latiusculum (Desv.) Underw. ex Hell. (고사리 전엽체의 증식에 미치는 배지구성물질과 배양방법의 영향)

  • Shin, So-Lim;Lee, Moo-Yeul;Choi, Jae-Sun;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.22 no.4
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    • pp.337-342
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    • 2009
  • Present studies were conducted to evaluate the effects of medium strength(MS and Hyponex), carbon sources and their concentrations, agar concentrations, and inoculation amounts on prothallus propagation of Pterdium aquilinum var. latiusculum(Desv.) Underw. ex Hell in vitro. The optimum MS medium strength for prothallus propagation was 2MS concentration. Phosphate source was most effective for prothallus growth of P. aquilinum var. latisculum. The addition of 1% sucrose or glucose to MS medium promoted prothallus multiplication. Growth of prothallus was not affected by agar concentration. Propagation of homogenized prothallus was vigorous even in liquid medium. Chopped gametophytes(100 and 200 mg) were inoculated on 250 ml ${\Delta}$flask with 100 mL of 2MS concentration medium and suspension culture was done at 100 rpm for 22 days. After 20 days, prothallus multiplication slowed down, so 100 mg of chopped prothalli is recommended for initial inoculation, since initial amount of inoculum did not affect subsequent prothallus multiplication. Consequently after 20 days of suspension culture, prothallus should be subcultured or transplanted outside of growing vessels.