• Korean Journal of Plant Tissue Culture
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• v.28 no.5
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• pp.239-242
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• 2001

The effects of post-cultural addition of liquid medium addition to stimulate in vitro bulblet growth in Lilium oriental hybrid 'Casa Blanca' were investigated. The sections of bulblets with swollen basal plate (7 mm$\times$12 mm) were cultured on medium containing 60 g/L sucrose and 1 g/L activated charcoal for two months in dark, and then, liquid medium was added into the same vessels. The addition of liquid medium stimulated the growth of bulblets remarkably, compared to no addition of liquid medium. The liquid medium supplemented with 120 g/L sucrose and double strength of MS salts were the most effective on the growth of in vitro bulblets.

• Journal of Life Science
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• v.20 no.9
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• pp.1402-1408
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• 2010

The effects of rice hull (RH) powder on the anticarcinogenic activity of submerged-liquid cultures of Agaricus blazei Murill (AB) were assessed for mouse ascites cancers induced by mouse Sarcoma S-180 (S-180) cancer cells. Optimal growth of AB mycelia in the basal liquid culture medium, containing soybean meal, was achieved by culturing at $25^{\circ}C$ for 5 days, when evaluated by $\beta$-glucan content, Brix, and mycelial weight, relative to other culture conditions. Hot-water extract (HWE) of the submergedliquid culture of AB mycelia grown at $25^{\circ}C$ for 5 days exhibited a stronger anticarcinogenic activity, relative to HWE from other culture conditions. No such effects were obtained from AB mycelial cultures by alternative temperature-controlling cultures. Both cytotoxicity for S-180 cells and anticarcinogenic potentials for mouse ascites cancer of the HWE from AB mycelia grown in the basal medium containing 1% RH powder for 5 days at $25^{\circ}C$ were significantly (p<0.05) enhanced, relative to HWE from the AB mycelia culture of the basal medium without RH powder. These results indicate that HWE of submerged-liquid culture of AB mycelia, incubated in media containing 1% RH powder at $25^{\circ}C$ for 5 days, enhanced anticarcinogenic activity against S-180 cell-induced mouse ascites cancer, and suggest that RH powder is an excellent ingredient for the improvement of the anticarcinogenic potentials of the submerged-liquid culture of mushroom mycelia.

• Applied Biological Chemistry
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• v.19 no.4
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• pp.219-226
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• 1976

To meet the need of protein feed and fine more efficient ways of returning waste to resources, we have carried out the study of the production of yeast for foods and feeds from the corn starch cake. The present study includes the method for acid-hydrolysis, the selection of yeast capable of utilizing hydrolyzate of the corn starch cake, and culture condition of Candida tropicalis under the liquid culture and the semisolid culture. Obtained results were as follows. 1. Hydrochloric acid was more excellent on the hydrolysis of the corn starch cake than sulfuric acid, and the yield of sugar was maximum, 57.2%, when the corn starch cake was hydrolyzed with 1.0% of hydrochloric acid at 2.0kg/cm for 30 minutes. 2. As the acid solution content was increased, more sugar was liberatedfrom the mixture, until the acid solution-substrate ratio reached 10:1. Beyond this point, no further increase was observed. To prepare the cultural medium of semisolid fermentation, a acid solution to substrate ratio of 3:1 appeared to be optimum. 3. Out of 6 yeast strains, Candida tropicalis had excellent growth on the hydrolyzate of the corn starch cake, and optimum temperature and initial pH were $30^{\circ}C$ and 6.0 respectively. 4. Optimum liquid medium of Candida tropicalis is ures 0.3%, potassium phosphate monobasic 0.15g and magnesium sulfate 0.04g in 100ml of the hydrolyzate of the corn starch cake, while optimum semisolid medium is ammonium chloride 0.4g, potassium phosphate monobasic 0.1%, magnesium sulfate 0.04%. 5. Candida tropicalis could assimilate the sugar in the hydrolyzate up to more than 88.75%, and a yield of dry yeast reached 19.13% to the corn starch cake under the liquid culture. 6. Compared to the that of the untreated corn starch cake, the cellulose content of the semisolid fermented cake decreased by 3.76% to 14.7%, whereas dry yeast contents increased by 13.89%.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.367-374
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• 2000

The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

• KSBB Journal
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• v.13 no.2
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• pp.155-161
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• 1998

The hairy roots of Crotalaria sessiliflora were induced from the tissue segments infected with Agrobacterium rhizogenes ATCC 15834. The induced hairy roots were subjected to paper electrophoresis fro the detection of opine-positive clones which were considered to have been transformed. Mannopine and agropine were presented in hairy root clones while mannopine was presented in two hairy root clones. Eight hairy root clones were selected and cultured in MS, B5 and WP media. Each of hairy root clones was showed a difference in branch pattern and growth rate. The best culture medium and culture conditions of hairy roots were in $\frac{1}{2}$MS(3% sucrose, pH 5.7) liquid medium at 25$^\circ C$, 70 rpm under dark, the growth rate in $\frac{1}{2}$MS liquid medium was increased with 210-fold more than that of inoculated hairy roots and with 2-fold more than that in MS liquid medium. Also, the adequate condition for hairy root growth was such that concentration of KH$_2PO$_4 was 1.25mM and the ratio of NH${_4}{^+}$ : NO${_3}{^-}$ was 1 to 3 in MS medium. The presence of pyrrolizidine alkaloids, monocrotaline, in the hairy roots was detected by TLC.

• KSBB Journal
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• v.18 no.6
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• pp.435-439
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• 2003

Plant cell culture can be divide into two classes non-organic culture and organic culture. Non-organic culture such as suspension culture has many researches, however organic culture about recombinant protein production has little researches. Recombinant protein produced through organ culture is quite stable and it can make proteins by itself without any grow regulators. Therefore organ culture is much easier than other methods. In this research, we used transformed tobacco seed. At first we germinated the seed then separated stems and leaves from the grown plant. And raised in liquid medium by in vitro vegetative reproduction. Continuing most suitable conditions, we compared the Quantities of recombinant protein from intra cellular with from extra cellular. And adding some permeabilizing agents (Pluronic F-68, Triton X-100, DMSO, PEG8000), we increased the productivity of the recombinant protein.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.135-140
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• 2001

A series of studies were carried out to establish micropropagation system, using airlift bioreactors (ebb $\varepsilon$ flood type, 5 L), of Lilium oriental hybrid 'Casa Blanca'. The bulblets with swollen basal plate were formed from bulb scales, then proliferated to bulblet clusters with swollen basal plate. Finally normal bulblets were formed from the sections. Bulblet formation and proliferation with swollen basal plate were not accomplished entirely in liquid culture of 5 L airlift bioreactors, but leafy bulb scales grew vigorously. Bulblet clusters with swollen basal plate were proliferated by periodic immersion culture. Bulblet proliferation was not affected by light, but scale leaves grew under light. MS medium containing 2.0 mg/L benzyl adenine (BA) and 0.3 mg/L indole acetic acid (IAA) was favorable to the bulblet proliferation with swollen basal plate. In liquid culture of 5 L bioreactors, bulblets from bulblet sections with swollen basal plate grew vigorously on MS medium with 70 g/L sucrose. It was effective for bulblet growth to replace the new medium after 8 weeks in culture during 16 weeks of cultural period. 15 g injection of bulblet sections as a cultural material was suitable for bulblet growth in 5 L bioreactors.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.37-37
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• 2015

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

• The Korean Journal of Mycology
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• v.42 no.2
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• pp.97-103
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• 2014

A total of 7039 strains of filamentous fungi are preserved in Korean Agricultural Culture Collection (KACC). The 4065 strains (58%) of them, which produce many spores in cultivation on proper media, are preserved with freeze-drying method. They are also preserved with liquid nitrogen and deep-freezer storage in order to minimize loss by death. Aspergillus, Penicillium, Lichtheimia, Mucor, Rhizopus, etc. which are common in surrounding environments, are included in this category. The others which do not produce spores, or produce few spores in vitro, are preserved with liquid nitrogen, deep-freezer and mineral oil storage. Phytophthora, Pythium, Cercospora, Septoria, Rhizoctonia, etc. are included in this category. The authors also introduced various fungal preservation methods and provided detailed preservation procedures that are used in KACC.

• Journal of Life Science
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• v.14 no.4
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• pp.560-566
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• 2004

Agrocybe aegerita is Hymenomycetes fungus belonging to the order Agaricales and family of Bolbitiaceae. Much less is known about liquid culture of Agrocybe aegerita. Thus, the present study was to investigate the liquid cultural characteristics of Agrocybe aegerita my-celium. The optimal medium for the mycelial growth and density was ME medium, optimal tem-perature and initial pH were 25$\pm$1$^{\circ}C$ and 5.5, respectively. And optimal culture time for mycelial growth was 12 days. The modified optimal medium compositions were dextrin 3% (w/v), yeast extract 2% (w/w), MgSO$_4$0.05% (w/v), and KH$_2$PO$_4$0.15% (w/v). Under optimal culture conditions, the mycelial growth of modified optimal medium was higher than that of ME medium.