• 제목/요약/키워드: bottle culture

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Effects of Fruiting Productivity of Grifola frondosa Using Bottle Cultivation according to Different Substrate Composition (잎새버섯에서 배지조성이 병재배 자실체 생산성에 미치는 영향)

  • Kim, Jeong-Han;Jeon, Dae-Hoon;Kang, Young-Ju;Jeoung, Yun-Kyeoung;Lee, Yun-Hae;Chi, Jeong-Hyun
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.150-154
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    • 2016
  • To determine a favorable substrate formulation for Grifola frondosa, physicochemical conditions, culture properties, and yields according to various substrate formulations were investigated. Based on these analyses, T4 (80:5:15 ratio of oak sawdust to dried bean-curd refuse to corn husk) resulted in a shorter cultivation period and higher yields (weight of fresh mushrooms harvested at maturity) than those of other treatments. The physicochemical properties of T4 were pH 5.4, 2.4% crude fat contents, 54 C/N ratio, 74.3% porosity, and 0.26 g/cm3 bulk density. These results emphasize the importance of optimal substrate development on the production efficiency of G. frondosa mushrooms and have implications for commercial applications.

Effects of Excising In Vitro-Formed Roots on Acclimatization of Micropropagated Cassava Plantlets (카사바의 미세증식에서 기내 발생 부정근의 절단이 순화에 미치는 영향)

  • Yoon, Sil;Cho, Duck-Yee;Soh, Woong Young
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.103-108
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    • 2001
  • The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

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Effect of Lactobacillus mucosae on In vitro Rumen Fermentation Characteristics of Dried Brewers Grain, Methane Production and Bacterial Diversity

  • Soriano, Alvin P.;Mamuad, Lovelia L.;Kim, Seon-Ho;Choi, Yeon Jae;Jeong, Chang Dae;Bae, Gui Seck;Chang, Moon Baek;Lee, Sang Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.11
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    • pp.1562-1570
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    • 2014
  • The effects of Lactobacillus mucosae (L. mucosae), a potential direct fed microbial previously isolated from the rumen of Korean native goat, on the rumen fermentation profile of brewers grain were evaluated. Fermentation was conducted in serum bottles each containing 1% dry matter (DM) of the test substrate and either no L. mucosae (control), 1% 24 h broth culture of L. mucosae (T1), or 1% inoculation with the cell-free culture supernatant (T2). Each serum bottle was filled anaerobically with 100 mL of buffered rumen fluid and sealed prior to incubation for 0, 6, 12, 24, and 48 h from which fermentation parameters were monitored and the microbial diversity was evaluated. The results revealed that T1 had higher total gas production (65.00 mL) than the control (61.33 mL) and T2 (62.00 mL) (p<0.05) at 48 h. Consequently, T1 had significantly lower pH values (p<0.05) than the other groups at 48 h. Ammonia nitrogen ($NH_3$-N), individual and total volatile fatty acids (VFA) concentration and acetate:propionate ratio were higher in T1 and T2 than the control, but T1 and T2 were comparable for these parameters. Total methane ($CH_4$) production and carbon dioxide ($CO_2$) were highest in T1. The percent DM and organic matter digestibilities were comparable between all groups at all times of incubation. The total bacterial population was significantly higher in T1 (p<0.05) at 24 h, but then decreased to levels comparable to the control and T2 at 48 h. The denaturing gradient gel electrophoresis profile of the total bacterial 16s rRNA showed higher similarity between T1 and T2 at 24 h and between the control and T1 at 48 h. Overall, these results suggest that addition of L. mucosae and cell-free supernatant during the in vitro fermentation of dried brewers grain increases the VFA production, but has no effect on digestibility. The addition of L. mucosae can also increase the total bacterial population, but has no significant effect on the total microbial diversity. However, inoculation of the bacterium may increase $CH_4$ and $CO_2$ in vitro.

Culture conditions of liquid spawn and the growth characteristics of Pleurotus ostreatus (느타리버섯의 액체종균 배양 조건과 생육 특성)

  • Lee, Soo-Jung;Kim, Hun-Hwan;Kim, Seon-Ho;Kim, In-Soo;Sung, Nak-Ju
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.162-170
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    • 2018
  • To improve the productivity of Pleurotus ostreatus, different conditions of liquid spawn culture were tested. The optimum culture conditions were potato dextrose broth, incubation temperature of $22^{\circ}C$, and pH 6. Fifteen different media ('A' to 'O') containing 0.3% soybean meal (SM) were prepared by varying sugar and glucose contents. The cultures were propagated in SM media for 14 days, at $22^{\circ}C$, and pH 6. Their absorbances were higher after 14 days of incubation in the media containing both sugar and glucose. In particular, the absorbance of media containing 5 to 20% of glucose alone ('C', 'D', 'E', and 'F') tended to increase in the incubation period. Dry cell weight was lower in media containing less than 20% sugar or glucose alone than in media containing 30% sugar ('A') or 30% glucose ('B') alone. In sawdust media, in 900 mL-bottle, the optimum inoculation volume of liquid spawn was 15 to 20 mL. The texture of the mushroom cultivated with the liquid spawn was superior to that cultivated in the solid spawn.

Characteristics of mycelial growth and fruit body production in two strains of the genus Agrocybe ASI19003(A. cylindracea) and ASI19016(A. chaxingu) (볏짚버섯속 ASI1 9003(버들송이)과 ASI1 9016(차신고)의 배양 및 재배적 특성)

  • Cheong, Jong-Chun;Seok, Dong-Kwon;Kim, Seung-Hwan;Jhune, Chang-Sung;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.13-19
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    • 2008
  • This experiment was carried out to examine physiological and cultural characteristics of two strains ASI 19003, 'Poplar field-cap mushroom' Agrocybe cylindracea, and ASI 19016, 'Chaxingo' A. chaxingu, at the bottle cultivation which have very similar morphological characteristics in genus Agrocybe. There was significant difference between the physiological and cultural characteristics of ASI19003 and ASI19016. The optimal temperature for the hyphal growth was $28^{\circ}C$ in the strain ASI19003 and $30^{\circ}C$ in ASI19016. The optimal pH was not different in two strains and these strains grew well at pH 5.5~7.0. But the optimal pH in the submerged culture was 5.5 in ASI19003 and 5.0 in ASI19016. Especially, hyphal growth of the strain ASI19016 was very poor at pH 6.0~7.3. The optimal carbon source for the growth was lactose in the strain ASI19003 and fructose in ASI19016, and nitrate sources were asparagine, alanine, and glycine in the strain ASI19003, and ammonium tartrate, asparagine, glycine, and alanine in ASI19016, respectively. The periods of incubation and fruiting body formation in the bottle cultivation during the spring were 27 and 13 days in the strain ASI19003, 29 and 17 days in ASI19016. The yields of fruit body were 114 g per bottle (850 $m{\ell}$ volume) in the strain ASI19003 and 100 g in ASI19016. In the summer, the periods of hyphal incubation and fruiting body formation were 29 and 11 days in the strain ASI19003, 30 and 12 days in ASI19016. The color of the cap in the ASI19003 strain according to temperature increase during the fruit body development become more pale, but the strain ASI19016 kept dark color relative to ASI19003. The fruiting body formation of the strain ASI19016 was faster than that of ASI19003. Accordingly, the cultivation of A. cylindracea ASI19003 during the spring, fall and winter, and A. chaxingu ASI19016 during the summer can keep high quality and stable supply all year round of these mushrooms.

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Acquisition of Thermotolerance in Transgenic Orchardgrass Plants with DgHSP17.2 Gene

  • Kim, Ki-Yong;Jang, Yo-Soon;Cha, Joon-Yung;Son, Daeyoung;Choi, Gi Jun;Seo, Sung;Lee, Sang Jin
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.5
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    • pp.657-662
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    • 2008
  • To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, the recombinant DgHSP17.2 gene was introduced into orchardgrass plants using the Agrobacterium-mediated transformation method and expressed constitutively under the control of the CaMV 35S promoter. The results of genomic DNA PCR and Southern analysis showed a DNA band and hybridization signal on agarose gel and X-ray film in transgenic orchardgrass plants harboring the recombinant DgHSP17.2 gene, but a DNA band and hybridization signal were not observed in the wild type and empty vector control plants. The same result was also obtained in RT-PCR and Southern blot analysis, and these transgenic orchardgrass plants did not show any morphological aberration both in the culture bottle and soil mixture. When leaf discs cut from transgenic orchardgrass plants with recombinant DgHsp17.2 gene were exposed to lethal temperature (heat treatment at $60^{\circ}C$ for 50 min), 60-80% of the leaf discs showed only damage symptoms, but non-transgenic leaf discs showed a lethal condition. These results indicate that the DgHsp17.2 gene may act as a protector from heat stress in plants.

Cultural and Morphological characteristics of fruit body of genus Agrocybe on Artificial Cultivation in Summer (볏짚버섯속(Agrocybe) 보존균주의 여름철 인공재배시 재배 및 형태적 특성)

  • Cheong, Jong-Chun;Seok, Dong-Kwon;Kim, Seung-Hwan;Jhune, Chang-Sung;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.7 no.1
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    • pp.1-8
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    • 2009
  • Thirty strains of genus Agrocybe which had been kept in National Institute of Horticultural and Herbal Science, were tested for the formation of fruiting body. Nineteen strains of collected genus Agrocybe were formed fruiting body at bottle culture of sawdust medium. Five strains (including ASI 19003) and thirteen strains (including ASI 19007) could be grouped as A. cylindracea and A. chaxingu. Although ASI 19008 showed the formation of fruiting body, morphological characteristics were significantly different from the two groups. Cultural period of ASI 19003 strain at sawdust substrates was 29 days and ASI 19007 train was 30 days in liquid spawn inoculation. The yield and quality of ASI 19003 strain was excellent in the spring, fall, and winter, whereas ASI 19007 strain was excellent in the summer. Accordingly, these cultivar might be contribute to farmers' income by stable year-round production if using the season-oriented strain.

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Bioconversion of Linoleic Acid to Conjugated Linoleic Acid by Bifidobacterium breve

  • Song Yoon-Seok;Kang Seong-Woo;Oh Deok-kun;Rho Yong-Taik;Hong Suk-In;Kim Seung-Wook
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.357-361
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    • 2005
  • The bioconversion of linoleic acid (LA) to conjugated linoleic acid (CLA) was investigated to examine LA-adaptation of Bifidobacterium breve KCTC 3461 to additions of 1 to 5 mg/mL of LA overtime. To induce LA-adaptation, B. breve KCTC 3461 was treated with LA, according to three schemes. For LA-adapted B. breve the maximum concentration of CLA, $300\~350{\mu}g/mL$, was obtained in cys-MRS medium containing 1 mg/mL of LA. The CLA production significantly increased with increasing LA concentration, from 1 to 4 mg/mL, but the conversion of LA to CLA gradually decreased. The CLA production capability of B. breve, and its tolerance, improved significantly with LA-adaptation. The addition of LA (1 mg/mL) into the culture broth after 24 h of cultivation in a 100-mL media bottle was most effective at promoting CLA production. In a 2.5-L stirred-tank bioreactor, the observed conversion and productivity of $56.6\%\;and\;35.4{\mu}gmL^{-1}h^{-1}$, respectively, by LA-adapted B. breve were approximately 6.6 and 9.8 times higher than those of LA-unadapted B. breve.

Identification of bacterial contaminants in porcine semen and its removal (돼지 정액내의 오염 세균의 동정 및 오염된 세균의 제거)

  • Park, Choon-Keun;Hong, Ki-Hun;Lee, Yong-Seung;Hahn, Tae-Wook;Son, Su-Jung
    • Korean Journal of Veterinary Service
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    • v.31 no.4
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    • pp.547-554
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    • 2008
  • Bacteriospermia is a frequent finding in fresh boar semen and can result in detrimental effects on semen quality and longevity. The objectives of this study was to evaluate types of bacterial contaminants in porcine fresh semen and the reducing effect of antibiotic and density gradient with percoll on the bacterial contaminants. Fresh semen was collected by gloved-hand method into a pre-warmed($37^{\circ}C$) thermostable bottle, and was inoculated onto blood agar and MacConkey agar, respectively. After incubated for 48 hour, 7.5% $CO_2$ at $37^{\circ}C$, bacterial colonies were selected and identified by Gram staining, oxidase test, catalase test and finally identified using API kits and Vitek system. Aerobic culture yielded a variety of bacteria from different genera. The most prevalent contaminant of fresh semen were Leclecia adecarboxylata, Acineobacter banmanni, Staphylococcus epidermidis, Staphylococcus cohni spp urealyticus, Proteus mirabilis. Most of identified bacteria were Gram(-) and non-pathogenic bacteria. It seems that bacterial contaminants in fresh semen were seem originated from multiple sources at the stud/farm, and were from animal and non-animal origins. Gentamicin treatment did not eliminate the bacterial contaminants completely but 3 step-density gradient with percoll completely removed the bacterial contaminants in fresh semen. Therefore, future study is necessary to prove that density gradient method with percoll can eliminate bacteria in fresh semen without significantly affecting sperm viability or function.

Somatic embryogenesis and plant regeneration in elite genotypes of Picea koraiensis

  • Li, Cheng-Hao;Liu, Bao-Guang;Kim, Tae-Dong;Moon, Heung-Kyu;Choi, Yong-Eui
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.259-265
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    • 2008
  • Picea koraiensis, called Korean spruce, is an evergreen tree and found mostly in northeast Asia. In this study, plant regeneration via somatic embryogenesis from open-pollinated immature zygotic embryos of nine geno-types of elite trees was established. Immature zygotic embryos were cultured onto RJW medium modified from 505 medium with $21.48{\mu}M$ NAA, $2.22{\mu}M$ BA, and $2.32{\mu}M$ KT. The average frequency for all nine genotypes was 74.2%. Embryogenic calluses of the nine genotypes of elite trees were subcultured on RJW basal medium containing $8.06{\mu}M$ NAA, $1.11{\mu}M$ BA, and $1.16{\mu}M$ kinetin. The calluses of three lines, $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$, were actively proliferated but others were not. Somatic embryogenesis was induced from the embryogenic callus in genotypes of $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$ on RJW medium with ABA and $60g\;l^{-1}$ sucrose. Cotyledonary somatic embryos were subjected to a drying process. The drying of embryos by uncapping the culture bottle for 5 days on a clean bench resulted in a high frequency of germination of somatic embryos (87% in RJW medium). However, plantlet conversion from germinated embryos was greatly reduced and the optimal medium for plant conversion was 1/2 WPM or 1/2 BMI medium. In conclusion, we have, for the first time, established a plant regeneration system via somatic embryogenesis in the Korean spruce, which can be applied for rapid micropropagation of elite trees.