• Title/Summary/Keyword: in vitro cultivation

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Conditions of In Vitro Spore Germination and Prothallium Culture for Sporophyte propagation of Polystichum braunii (Spenn.) Fée (좀나도히초미(Polystichum braunii (Spenn.) Fée) 포자체 증식을 위한 기내 포자 발아와 전엽체 배양 조건)

  • Kwon, Hyuk Joon;Han, Ji Hyun;Lee, Cheol Hee;Kim, Soo-Young
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.454-461
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    • 2017
  • This study was conducted to investigate the optimal conditions for spore germination, prothallus propagation, sporophyte formation, and seedling growth in Polystichum braunii (Spenn.) $F{\acute{e}}e$. The rate of spore germination and early prothalium development was high in Knop (41.2%), which had low mineral content. The optimal medium for prothallus propagation and sexual organ formation was 2MS medium (2% sucrose). Among the various mixtures of cultivation soil (bedding soil, peat moss, perlite, and decomposed granite), a mixture of bedding soil and decomposed granite at a ratio of 2:1 (v:v) had a positive effect on sporophyte formation (276.3 ea/$7.5m^2$). The most efficient conditions for promoting the growth of sporophytes were pots filled with only bedding soil.

Effect of TDZ (Thidiazuron) on Shoot Proliferation of Peace Poplar (Peace 포플러(Populus koreana X P. trichocarpa)의 줄기형성에 미치는 Thidiazuron 효과)

  • Kang, Ho-Duck;Moon, Heung-Kyu;Park, In-Sun;Lee, Min-Soon
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.49-53
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    • 2004
  • Shoot formation was investigated from in vitro cultivation of exotic hybrid poplar (Populus koreana ${\times}$ P. trichocarpa) with a specific stomatal character occurring both upper and lower surface of leaves. Two different explants (stem and leaf segment) of Peace poplar were cultured on half strength Murashige and Skoog (MS) basal medium supplemented with the various concentrations of thidiazuron as a plant growth regulator. Most adventitious shoots were produced from excised ends of stem or mid-veins of leaf segments. The highest average numbers of shoots were 7.1 and 5.3 with the treatments of 0.02mg/L TDZ in both explants of stem and leaf segment. The highest shooting rates were achieved to 83.3% and 47.6% with the concentrations of 0.01mg/L and 0.02mg/L TDZ by axillary bud and leaf cultures, respectively.

High Frequency Plant Regeneration from Leaf, Petiole and Internode Explants of Codonopsis lanceolata Benth.

  • Ghimire, Bimal Kumar;Shin, Chul-Min;Li, Cheng Hao;Kim, Na-Young;Chung, III-Min;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.73-81
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    • 2007
  • An efficient regeneration system was developed using leaf, petiole, and internode explants. Highly embryogenic callus was obtained following cultivation on MS basal nutrient supplemented with 2 $mg/{\ell}$ 2,4-D. Globular, heart, torpedo and cotyledon shaped somatic embryo were produced from the surface of embryogenic callus. Direct shoot regeneration without intermediate callus formation has been achieved on MS medium supplemented NAA and BAP. The percentage of response varies with different concentration of auxin and cytokinin treated individually or in combination. The best shoot regeneration response (54.28%) and number of shoot per explant (12.67) were achieved on the medium supplemented with 0.1 $mg/{\ell}$ NAA and 1 $mg/{\ell}$ BAP. The regenerated shoot transformed into young plant when cultured into elongation and root induction medium. More than 90% of in vitro propagated plants could survive when transferred to the greenhouse for acclimation. This optimized regeneration system can be used for rapid shoot proliferation and genetic transformation.

Experimental and epidemiological studies on the life cycle of Echinostoma hortense Asada, 1926(Trematoda: Echinostomatidae) (남한강류역(南漢江流域)의 호르텐스극구흡충(棘口吸蟲) 감염실태(感染實態)와 생활사(生活史)에 관(關)한 연구(硏究))

  • Ahn, Yung-Kyum;Ryang, Yong-Suk
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.121-136
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    • 1986
  • Recently there have been some reports on human infections of Echinostoma hortense in Korea. It was found that a few species of freshwater fishes were playing the role of the second intermediate host of E. hortense. However, molluscan intermediate host has not been identified yet in Korea. The present study aimed to establish the life cycle of E. hortense in laboratory. Experimental studies such as egg production from the rat, development of the eggs in vitro, exposure of miracidia to freshwater snails, shedding pattern of cercariae from infected snails, morphology of cercariae, cercarial infection to the second intermediate host and infection of metacercariae to the definitive hosts were done. In addition, epidemiological surveys on the infection status in inhabitants and house rats, and on the natural infection of larval echinostomes in the snails and fishes were carried out along the South Hangang-river. The results obtained were as follows: 1. The eggs deposited from adults in physiological saline were cultivated at room temperature($20{\sim}24^{\circ}C$). The miracidia were firstly observed on 8 days after cultivation, and 85.5% of the eggs contained the mature miracidia on 11 days after cultivation. More than 90% formed the miracidia when cultivated at temperature $22{\sim}27^{\circ}C$. Hatching of the miracidia began on 12 days after cultivation and continued for a week. The size of the miracidia was $103.0{{\times}}51.4{\mu}m$ in average. The motility of the miracidia were active up to 8 hours after shedding, but they were all dead within 10 hours after shedding. 2. A freshwater snail, Radix auricularia coreana was cultivated in aquaria. A hatched $F_1$ snails from the egg masses were exposed to 20 miracidia respectively. Escape of cercariae started on 15 days after infecton. Radix auricularia coreana was experimentally identified as the first intermediate host of E. hortense in Korea. 3. Cercarial shedding started on $15{\sim}20$ days after infection by snail, continued for about 10 days (8.8 days in average). Infected snails were dead within 32 days after the miracidial infection. About 1,335 cercariae($328{\sim}1,994$) per snail were shed in its life, and 119 cercariae in average per snail per day were shed. The cercariae were motile for more than 24 hours, and then squirming at the bottom until death. The body and tail sizes of cercariae were $356{\times}186{\mu}m$ and $510{\times}68{\mu}m$ in average, respectively. The rediae parasitized in the snail hosts were found mainly around the pericardial regions, and their size was $1,575{\times}258{\mu}m$ in average. The numbers of developing cercariae in a mature redia were 14 in average ($7{\sim}20$ in range). The numbers of rediae in a snail were 102 in average on 15 days after miracidial infection and 221 in average on 28 days. 4. Three uninfected Misgurnus anguillicaudatus, less than 6.5cm long were used in for the cercarial infection. They were all exposed with 755 cercariae, and examined at 5-day intervals starting from 10 days after infection. All the fishes were infected with metacercariae of E. hortense and a total of 275 was found infected(36.4%). The metacercariae were fed to rats and the adult worms were obtained on 15 days after infection. 5. The infected rats began to deposit the eggs on 11 days after infection. The number of eggs deposited per day per worm (EPD/worm) was $400{\sim}500$ on 3 weeks after infection and was increased to $1,000{\sim}1,500$ on 4 to 17 weeks, then decreased to 800 on 21 weeks after infection. 6. A total of 745 stool specimens collected from 576 male and 169 female residents of 8 different villages along South Hangang basin was examined. Out of 745 specimens, the eggs of Echinostoma sp. were found in 2 cases (0.3%). Of 34 house rats one showed egg-positive (2.9%). 7. Total 971 Radix auricularia coreana collected from 7 sampling stations were examined for shedding of cercariae. Three snails (0.3%) shed the cercariae of E. hortense. A total of 119 out of 542 freshwater fishes (22.0%) had the metacercariae of E. hortense. The fishes parasitized with the metacercariae were 4 out of 14 examined species. The infection rates of 4 species were 34.1% (106 out of 311) in Misgurnus anguillicaudatus, 30.4% (7 out of 23) in Misgurnus mizolepis, 4.3% (2 out of 46) in Moroco oxycephalus and 22.2% (4 out of 18) in Odontobutis obscura interrupta. In summarizing the above results, the first intermediate host of E. hortense was found as Radix auricularia coreana in Korea. Also, it took about 46 days for the shortest completion of a life cycle of E. hortense in summer; that is, 10 days for miracidial development in eggs, 15 days for cercarial development in the snail, about 10 days for metacercarial development in the second intermediate hosts, and 11 days for the maturation as the adults in the definitive hosts. The natural infection rates of E. hortense in the intermediate hosts were relatively high but those in the definitive hosts were low in the middle areas of South Hangang basin.

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Isolation and cultivation of follicle constituting cells from human hair follicles

  • Shin, Youn-Ho;Seo, Young-Kwon;Lee, Doo-Hoon;Yoo, Bo-Young;Song, Kye-Yong;Seo, Seong-Jun;Whang, Sung-Joo;Kim, Young-Jin;Yang, Eun-Kyung;Park, Chang-Seo;Chang, Ih-Seop;Park, Jung-Keug
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.365-368
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    • 2003
  • Hair follicles develop as a result of epithelial-mesenchymal interactions between epidermal keratinocytes and dermal cells. Moreover hair follicles constitute multiple cells that influence hair follicle development and cyclic activity. We isolated some cells using explantation and enzymatic digestion method from human scalp hair follicles. So we could culture some follicular cells, such as outer root sheath (ORS) cells, dermal papilla (DP) cells, dermal sheath (DS) cells, matrix cells and melanocyte.

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Physiological Studies on the Formation of Hairy Root by the A. rhizogenes. III. Attachment of A. rhizogenes strain A4 to Carrot(Daucus carota L.) Cells (Agrobacterium rhizogense에 Hairy Root 형성에 대한 생리학적 연구. III. 당근 세포에의 A. rhizogenes의 부착)

  • Hwang, B.;Hwang, S. J.;Ann, J. C.;Jo, H. S.
    • KSBB Journal
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    • v.4 no.2
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    • pp.94-98
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    • 1989
  • In vitro attachment experiments of bacteria to surface of host plant cell were carried out using C14 labeled cells of A. rhizogenes strain A4 and carrot protoplasts isolated from suspension culture of cells. Protoplasts were cocultivated with A. rhizogenes at various times after their isolation. Attachment kinetics showed that adherence of bacteria to protoplasts attained a maximum level within 120mins of co-cultivation. Maximum attachment occured at pH 6.0 and 24-35$^{\circ}C$. Bacterial attachment was observed at botg carrot cells with and without primary cell wall. The inhibition of transformation on the carrot root discs by A. rhizogenes was observed when non-related strain and heat inactivated bacterial strain cells were pretreated.

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Isolation and Identification of Competitive Fungi on Medium for Black Wood Ear Mushroom in Korea and In Vitro Selection of Potential Biocontrol Agents (목이버섯 배지 오염 곰팡이균의 분리, 동정 및 생물학적 방제제 선발)

  • Seoyeon Kim;Miju Jo;Sunmin An;Jiyoon Park;Jiwon Park;Sungkook Hong;Jiwoo Kim;Juhoon Cha;Yujin Roh;Da Som Kim;Mi jin Jeon;Won-Jae Chi;Sook-Young Park
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.66-77
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    • 2024
  • Black wood ear mushroom (Auricularia auricula-judae) is one of the most economically important mushrooms in China, Japan, and Korea. The cultivation of wood ear mushrooms on artificial substrates is more efficient in terms of time and cost compared with their natural growth on trees. However, if the substrate cultivation is infected by fast-growing fungi, the relatively slow-growing ear mushroom will be outcompeted, leading to economic losses. In this study, we investigated the competitive fungal isolates from substrates infected with fast-growing fungi for the cultivation of ear mushrooms in Jangheung and Sunchon, Korea. We collected 54 isolates and identified them by sequencing their internal transcribed spacer region with morphological identification. Among the isolates, the dominant isolates were Trichoderma spp. (92.6%), Penicillium spp. (5.6%), and Talaromyces sp. (1.8%). To find an appropriate eco-friendly biocontrol agent, we used five Streptomyces spp. and Benomyl, as controls against Trichoderma spp. and Penicillium spp. Among the six Streptomyces spp., Streptomyces sp. JC203-3 effectively controlled the fungi Trichoderma spp. and Penicillium spp., which pose a significant problem for the substrates of black wood ear mushrooms. This result indicated that this Streptomyces sp. JC203-3 can be used as biocontrol agents to protect against Trichoderma and Penicillium spp.

The Production of Anti-cancer Substances by in vitro Grown Cultures of Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Park, Kyung-Hwa;Kim, Yong-Hae;Yoon, Eui-Soo;Kang, Tae-Jin;Park, Kwang-Tae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 1999.10a
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    • pp.46-57
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    • 1999
  • Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

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Selection of Cultivars and Organic Solvents to Improve Fruit Set of Greenhouse Watermelon during Cold Period (저온기 시설수박 착과율 증진을 위한 품종 및 화분분리 유기용매 선발)

  • Lim, Chae-Shin
    • Journal of Bio-Environment Control
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    • v.19 no.3
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    • pp.147-152
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    • 2010
  • Poor fruit set during winter period is one of the biggest problem in plastic tunnel watermelon cultivation. Hand pollination is inevitable to maximize fruit set of the winter watermelon. Productivity and viability of pollen grain and organic solvents for pollen storage were investigated. All cultivars produced more than 10 mg/flower except for 'Kumchun' cultivar. Pollen amount per flower were 13.8 mg in 'Bok' and 12.1 mg in 'Speedkul'. Germination rate of pollen grains incubated at $30^{\circ}C$ right after soaking in pentane solvent were 76% in 'Kumchun' as the lowest and 92% in 'Apollokul' as the highest. The pollen of 'Bok' showed the highest germination rate by 75% after a 15-day storage in pentane. All cultivars showed their pollen germination rate below 25% after a 24-day storage. Among the cuitivars, speed of pollen tube growth in vitro were relatively lower in 'Kumchun' and 'Sambokkul' by below $50\;{\mu}m/hr$. Pollen tube of these cultivars tended to burst during its elongation on the medium. Pollen stored 24 hrs in organic solvents showed 45, 39, 34, 23, and 19% of germination in pentane, ethyl ether, n-hexane, ethyl acetate, and acetone, respectively. Compared with light condition, pollen viability was higher in darkness during pollen storage in organic solvents. Pollen grain was susceptible to the organic solvent. The viability of pollen grains seems to be influenced greatly by duration of soaking pollen in organic solvent and the polarity of solvents. Organic solvent damages surface of pollen grain and extent of damage was varied by the solvents.

Effect of MS Medium Strength on the Sprouting Rate and Growth Characteristics in Meristem Culture of Strawberry 'Seolhyang' ('설향' 딸기의 생장점 배양 시 MS 배지 농도에 따른 발아율 및 생육특성)

  • Kim, Hye Jin;Lee, Jong Nam;Lim, Hak Tae;Yeoung, Young Rok
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.100-104
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    • 2014
  • This study was conducted to determine the optimal MS medium strength to improve sprouting rate of apical meristem of strawberry 'Seolhyang' in vitro. Strawberry apical meristems at size (0.2 mm to 0.3 mm) with leaf primordials were cultured on the MS media with four strength levels, ($1/4{\times}$, $1/3{\times}$, $1/2{\times}$, and $1{\times}$) and the sprouting rate and growth characteristics were evaluated after eight weeks after cultivation. Shoot rate of 'Daewang' apical meristems was 93.6%whereas 'Seolhyang' apical meristems were sprouted with 31.6% on $1{\times}$ MS medium strength. Different sprouting rates were observed in 'Seolhyang' apical meristem with 31.6% in $1{\times}$ medium, 75.0% in $1/2{\times}$ medium, and 94.4% in $1/3{\times}$ medium. The sprouting rate was improved with the decrease of medium strength, but the shoot rate in $1/4{\times}$ medium decreased up to 54.5%. Shoot length was 0.9 cm in $1{\times}$ medium, 1.2 cm in $1/2{\times}$ medium, 1.6 cm in $1/3{\times}$ medium, and 1.9 cm in $1/4{\times}$ medium. Shoot length was longer as medium strength decreased and numbers of leaves and roots were not significant differences among the medium strengths. As a result, sprouting rate was highest and plant growth was best in $1/3{\times}$ MS medium compared to the others.