• Title/Summary/Keyword: Liquid medium

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Cryopreservation of in Vitro Grown Shoot Tips of Sweet Potato (Ipomoea batatas L.) by the Encapsulation-Vitrification Method

  • Yi, JungYoon;Lee, GiAn;Lee, YoungYi;Gwag, JaeGyun;Son, EunHo;Park, HongJae
    • Korean Journal of Plant Resources
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    • v.29 no.6
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    • pp.635-641
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    • 2016
  • Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulation-vitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at $25^{\circ}C$. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

Cryopreservation of Embryogenic Callus in Sweetpotato cv. 'Yulmi' (고구마품종 '율미' 배발생 캘러스의 초저온 동결보존)

  • Park, Jong-Suk;Kim, Suk-Weon;In, Dong-Su;Eun, Jong-Seon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.109-113
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    • 2003
  • Cryopreservation of embryogenic callus derived from apical meristem culture was attempted by slow prefreezing method (two-step method) with various cryoprotectants in sweetpotato cv. 'Yulmi' Precultured embryogenic calli on medium containing 10 mg/L ABA prior to slow prefreezing in liquid nitrogen indicated higher survival rate than 1.0 mg/L ABA preteatment. The cryoprotectant comprising 1.28 M DMSO in 0.4 M sucrose solution gave the best survival (over 46%) of sweetpotato cells exposed to liquid nitrogen as determined by TTC reduction and FDA staining method. Cryopreserved calli cultured on MS medium with 1.0 mg/L 2,4-D were grown for 4 weeks in the dark and induced embryos after another 4 weeks. They were subcultured on MS medium supplemented with 0.1 mg/L 2,4-D+0.1 mg/L kinetin for 2 weeks and regenerated into normal plantlets in MS basal medium.

Occurrence of Phytophthora Rot of Strawberry Caused by Phytophthora nicotianae var. nicotianae (Phytophthora nicotianae var. nocotianae에 의한 딸기 역병의 발생)

  • 송주희;노성환;하주희;정연화;문병주
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.445-451
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    • 1998
  • A severe Phytophthora rot of strawberry caused by a species of Phytophthora has been widely occurred at major cultivation areas of Kimhae on August in 1997. Incidence of the disease was obtained in the range of 69.2~83.6% in surveyed 4 fields and showed an average of 75.2%. A species of Phytophthora was mostly isolated from the crown of infected strawberry plants and all the isolates were identified as P. nicotianae var. nicotianae (=P. parasitica). The fungus showed strong pathogenicity on strawberry by inoculation test. As a result of the leaf inoculation using mycelial disks of the fungus, both leaves and petioles were darkly browned, and were finally blighted. As a result of the root inoculation of zoospore suspension, both roots and crowns were rotten with dark brown. Although the fungus produced sporangia either on V-8 juice agar medium or liquid medium, the sporangia observed on the liquid medium appeared to be broadly turbinate and noncaducous. Moreover the fungus cultured on the liquid medium often produced sporangia having two papilla. The number of zoospores in sporangia was found to be ranged from 3 or 4 to as many as 20 or 25. In addition, the released zoospore from the sporangium became the cystospore during the prolonged culture of the fungus. The sporangia were measured as av. 49$\times$35 ${\mu}{\textrm}{m}$ with l/b ratio of 1.43. All isolates from crowns were heterothallic and A1 mating type since oospores were abundantly formed on clarified V-8 juice agar by dual culture with P. capsici A2 mating type. Aplerotic oospores were sized 24-26 ${\mu}{\textrm}{m}$. Antheridia were always amphigynous and recoreded an average of 12$\times$10 ${\mu}{\textrm}{m}$. Hyphal swlling were easily observed, and terminal or intercalary chlamydospores were abundantly formed on V-8 juice agar as well as in C/Z solution and sized av. 28.2 ${\mu}{\textrm}{m}$. This is the first report of Phytophthora rot of strawberry in Korea.

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Production of Amylases from Herpetosiphon geysericola (Herpetosiphon geysericola 균주의 Amylase 생성)

  • Jun, Yeong-Soo;Seu, Jung-Hwn
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.188-191
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    • 1985
  • A thermophilic and cellulolytic bacterium, Herpetosiphon geysericola CUM 317 isolated from the compost, produced ${\alpha}-amylase,\;{\beta}-amylase$, and glucoamylase. Mutual relationships on the production of the three amylases were studied by changing the cultivation conditions. ${\alpha}-Amylase$ and glucoamylase were produced highly after 40 hrs on wheat bran medium at $50^{\circ}C$ and after 30 hrs on liquid medium at $40^{\circ}C$, though ${\beta}-amylase$ was produced best at 10 hrs of initial cultivation phase. The production of the amylases was generally repressed by the addition of carbon sources in liquid medium containing polypeptone. ${\alpha}-Amylase$ production was enhanced relatively by the addition of cupric sulfate in the liquid medium, ${\beta}-amylase$ was enhanced by cadmium sulfate, and glucoamylase was enhanced by calcium chloride.

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A Successful Regeneration from Shoot Tips of Chrysanthemum morifolium (Ramat.) following Cryopreservation by Droplet-vitrification

  • Yi, Jung-Yoon;Balaraju, Kotnala;Baek, Hyung-Jin;Yoon, Mun-Seop;Kim, Haeng-Hoon;Lee, Young-Yi
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.675-683
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    • 2018
  • This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of Chrysanthemum morifolium (Ramat.) cvs. 'Borami' and 'Yes morning'. The shoot tips of Chrysanthemum were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7 M). Precultured explants were treated with loading solution (LS, C6) containing glycerol 20% and sucrose 20% for 30 min and exposed to dehydration solution (B5) containing 40% of glycerol and 40% of sucrose for 60 min at $25^{\circ}C$, and then transferred onto droplets containing $2.5{\mu}l$ PVS3 on sterilized aluminum foils ($4cm{\times}0.5cm$) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposing of shoot tips to MS + 0.3M sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at $25^{\circ}C$ in both the cultivars. The viability of cooled samples, followed by culturing on $NH_4NO_3$-free MS medium for first 5 days was increased to two-fold (80.7%) regrowth rate over those cultured on normal MS medium or MS medium containing plant growth regulators. This result shows droplet-vitrification would be a promising method for cryobanking chrysanthemum germplasm.

Anticancer and Antioxidant Activities of Coriolus versicolor Culture Extracts Cultivated in the Citrus Extracts. (감귤 농축액에서 배양한 운지버섯 배양추출물의 항산화 및 항암활성)

  • 이세진;문성훈;김택;김진용;서정식;김대선;김율리아;김영준;박용일
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.362-367
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    • 2003
  • Coriolus versicolor was grown in a defined synthetic liquid medium and citrus extracts, and the culture extracts were examined for antioxidant activity, nitrite scavenging activity, and in vitro anticancer activity against HeLa, PC-3, HepG2, and A-549 cells. Whereas the culture extracts obtained from the synthetic medium and the un-inoculated citrus extract showed 60 and 22% of the 1,1-diphenyl-2-picrylhydrazyl radical scavenger activity, the culture extracts obtained from the citrus extracts medium exhibited antioxidant activity up to 89%. The nitrite scavenging activity of the culture extracts obtained from the citrus extracts medium and the synthetic liquid medium, and the un-inoculated citrus extract at pH 1.2 were up to 67, 55, and 34%, respectively. The culture extract obtained from the synthetic liquid medium inhibited the growth of HeLa, PC-3, HepG2, and A-549 up to 66, 23, 18, 10% at 48 h of incubation, respectively; however, the culture extract obtained from the citrus extracts medium inhibited the growth of HeLa, PC-3, HepG2, and A-549 up to 75, 82, 55, and 82%, respectively. As a negative control, the un-inoculated citrus extract was examined in the same way and inhibited the growth of HeLa, PC-3, and HepG2 cells 20, 6, and 15% at 48 h incubation, respectively; the inhibition of A-549 cell growth was negligible. These results clearly showed that the fermentation of C. versicolor in the citrus extracts rather than in the defined synthetic medium significantly enhanced the anticancer activity, antioxidant activity, and nitrite scavenging activity.

Determination of the Optical Functions of Various Liquids by Rotating Compensator Multichannel Spectroscopic Ellipsometry

  • Bang, Kyung-Yoon;Lee, Seung-Hyun;Oh, Hye-Keun;An, Il-Sin;Lee, Hai-Won
    • Bulletin of the Korean Chemical Society
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    • v.26 no.6
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    • pp.947-951
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    • 2005
  • Rotating compensator multichannel spectroscopic ellipsometry has been employed to determine the optical functions of various liquids used in chemistry. We attempted three different measurement configurations: (1) air-liquid interface, (2) prism-liquid interface, and (3) liquid-sample interface. In prism-liquid interface, we found that the prism surface had roughness and it should be considered in analysis for accurate results. In liquidsample interface, we had much higher reflection, better sensitivity, and less limitation compared to the other two configurations when crystalline silicon was used as reference sample. We discuss the merit of each configuration and present the optical functions of various liquids. Also we demonstrate Bruggeman effective medium theory to determine the optical properties of mixed liquid.

Changes of Nutrients in Media and Mycelia on Liquid Spawn Culture of Lentinula edodes (표고 액체종균 배양시 배지와 균사체의 양분변화)

  • Shim, Kyu-Kwang;Yoo, Young-Jin;Koo, Chang-Duck;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.42 no.2
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    • pp.144-149
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    • 2014
  • Lentinula edodes liquid spawn growth under explosive aeration (supplying air with tiny bubbles) and soybean meal addition to liquid culture medium were investigated in terms of mycelial growth and residual free sugar content. The two treatments were effective for homogeneous culturing of mycelial spawn and for separating colonies during multiplication after an exponential growth period without limiting sustaining nitrogen nutrients. The mycelial growth and carbon dioxide concentration were greatest on the 13th day since the inoculation. At 12th day, however, free sugars were almost depleted in the upper part of the liquid medium. Total nitrogen content within precipitated mycelia was the highest at the 13th day. Chitin and sucrose contents in the mycelia were the highest at the 18th day, but ergosterol content became highest at 22 days. These results suggest that Lentinula edodes liquid spawn is ready in 18 days after inoculation.

Studies of application of artificial ground freezing for a subsea tunnel under high water pressure - focused on case histories - (고수압 해저터널 건설을 위한 동결공법 적용성에 관한 연구 - 사례를 중심으로 -)

  • Son, Young-Jin;Lee, Kyu-Won;Ko, Tae Young
    • Journal of Korean Tunnelling and Underground Space Association
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    • v.16 no.5
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    • pp.431-443
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    • 2014
  • In this paper case studies of artificial ground freezing, which have not been applied in Korea, have been investigated for the water cut-off in a subsea tunnel under high water pressure and the most commonly used cooling mediums of brine and liquid nitrogen are examined. Since sea water with pressure has the lower freezing point than pure water, the lower temperature cooling medium is required in the application of subsea tunnel. Also, the cooling medium must have refrigeration safety and is able to reduce executing time. Brine freezing system can reuse cooling medium and is safer than liquid nitrogen freezing. But it takes more time to freeze ground and needs complex circulation plants. On the other hand, liquid nitrogen freezing system can't recycle cooling medium and may cause breathing problems or asphyxiation through oxygen deficiency. But, freezing with liquid nitrogen is fast and requires simple refrigeration equipment. Principal elements of design for ground freezing in subsea tunnel have been extracted and these elements are needed further research.

High Frequency Plant Regeneration in Embryogenic Cell Suspension Cultures of Cucumber (오이 배발생세포의 현탁배양을 통한 고빈도 식물체 재분화)

  • 정원중;우제욱;박효근;최관삼;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.289-291
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    • 1999
  • Hypocotyl explants from 7 days old seedlings of one $F_1$ hybrid cultivar and two pure lines of cucumber formed embryogenic calli at frequencies of up to 8% when cultured on Murashige and Skoog medium (MS) supplemented with 1 mg/L 2,4-D for 3 weeks. Embryogenic calli gave rise to somatic embryos. When slices of somatic embryos were cultured on the same medium for 4 weeks, they formed embryogenic calli. Embryogenic cell suspension cultures were established with embryogenic calli in MS liquid medium with 1 mg/L 2,4-D. Embryogenic potential of cell suspension cultures was maintained by subculturing every seven days. When the level of 2,4-D in the medium was lowered to 0.2 mg/L by diluting with liquid MS basal medium, embryogenic cell suspension cultures underwent development into numerous somatic embryos. When plated onto MS basal medium, over 95% of somatic embryos developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.

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