• Title/Summary/Keyword: 기내배양

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Shoot Regeneration from Leaf Explants of Digitalis purpurea L. (디기탈리스의 잎 절편으로부터 신초의 재분화)

  • Hwang, Sung-Jin;Lee, Hye-Jung;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.397-401
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    • 2003
  • Digitalis purpurea L. is a medicinal herb and have been used to congestive heart failure, mycocardial infarction, edema, angina etc. A protocol has been developed for in vitro propagation of adventitious shoot buds directly from leaf segments of D. purpurea Leaf explants of D. purpurea directly formed shoot buds when cultured on a MS medium supplemented with $2\;mg/l$ BA and $0.1\;mg/l$ IAA for 5 weeks. Adventitious shoots were multiplied by subculturing on the $B_5$ medium and shoot elongation was developed by subculturing on the WPM medium. Root formation from the shoot regenerated was achieved on MS basal medium containing 1 mg/ IBA.

Effect of MS Medium Strength and Nitrogen Concentration on Bulblet Formation and Growth of Lilium In Vitro (MS 배지 내 무기물 농도 및 질소 농도가 나리의 자구 형성과 비대에 미치는 영향)

  • 염미란;박현춘;양찬석;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.341-346
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    • 2001
  • To investigate the effects of MS medium strength and nitrogen concentration on bulblet formation from bulblet scale segment culture and bulblet growth from bulblet of Lilium oriental hybrid 'Casa Blanca', asiatic hybrid 'Mona', and longiflorum hybrid 'Hinomoto', 0.5∼2.0 strength of MS salts and 30∼120 mM nitrogen concentrations of MS medium were examined in vitro. The number of bulblets from bulb scale segment was favored in the strength of 0.5∼1.0 strength of MS salts or 30 mM total nitrogen concentration of MS medium in three cultivars. But the growth of bulblets formed in vitro was promoted in the 2 strength of MS medium or hight concentration of total nitrogen of MS medium up to 120 mM in three experimented cultivars.

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An Effective Method of Diagnosis of Potato Leafroll Virus by RT-PCR (RT-PCR 방법을 이용한 효과적인 감자 잎말림 바이러스의 검정)

  • Jeon, Jae-Heung;Joung, Young-Hee;Choi, Kyung-Hwa;Kim, Hyun-Soon;Oh, Hyun-Woo;Park, Se-Won;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.12 no.3
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    • pp.358-362
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    • 1996
  • 감자 잎말림 바이러스를 검정하기 위하여 ELISA 및 전자현미경에 의해 바이러스 감염이 확인된 기내 배양중인 감장의 줄기로부터 RT-PCR 분석을 수행하였다. 분리된 총 RNA들로부터 바이러스 cDNA를 합성하고 감자 잎말림 바이러스 외피단백질의 일부인 465bp를 특이하게 증폭하도록 고안한 두 primer를 사용하여 PCR 반응을 하였다. 증폭된 465pb의 DNA 절편의 염기서열을 분석한 결과 역시 감자 잎말림 바이러스임을 확인하였다. 바이러스 검정에 있어서 EL-ISA 방법과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법이 ELISA 방법보다 감자 잎말림 바이러스검정에 있어서보다 정확한 방법인 것으로 사료된다.

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In vitro Propagation of Trichosanthes kirilowii Max. (조직배양에 의한 하늘타리의 증식)

  • Hwang, Sung-Jin;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.1
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    • pp.15-20
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    • 2001
  • Adventitious shoot buds, without an intervening callus phase, were induced from stem explants of Trichosanthes kirilowii on MS medium supplemented with 2 mg/L BA. The culture medium, carbon source as well as plant growth regulators were found to influence further shoot multiplication and eventual shoot growth. A maximum number of shoots was obtained on a MS medium containing 0.5 mg/L BA and 3% sucrose. Shoot produced in vitro were rooted on half-strength phytagelled 1/2MS medium prior to transfer to green house condition.

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Ultrastructure of the Adventitious Root Meristem and Callus Induced by Tissue Culture of Tobacco(Nicotiana tabacum)Leaves (담배잎의 기내 배양에서 유기된 부정근 분열조직 및 캘러스 세포의 미세구조)

  • 차현철;박호일
    • Journal of the Korean Society of Tobacco Science
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    • v.17 no.1
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    • pp.33-40
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    • 1995
  • Structures of the adventitious root meristem induced from callus culture of tobaco (Nicotiana tabacum cv. NC 82) leaves were investigated by light and transmission electron microscopy. Structural differences between in vitro root and callus cells were also examined by the microscopy. The submicroscopic features of the in vitro root cells were as follows. Intercellular spaces were not developed and nuclei with two nucleoli were observed occasionally. Plasmodesmate were found in groups or sing1y on transverse and longitudinal walls. Amyloplast solely filled with starch grains, with one to five electron - dense bands, was surrounded by single membrane. in the callus cells, vacuolization of central part in the cytoplasm having mitochondria with swollen cristae and starch grains like those of in vitro root cells was a distinct feature. Vesicles which were found between cell wall and plasma membrane may be arisen by a process of protoplasmic invagination. By comparing of ultrastructures between the cells of callus and in vitro roots we found that the distinct differences lied on thickened cell walls and hypertrophed vacuoles in the former, and less thickened cell walls and several small vacuoles in the later.

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Influence of Growth Regulators on Adventitious Shoot Regeneration from Tissues of Malus domestica cv. 'Gala' in vitro (기내 사과나무 조직의 신초 재분화에 미치는 생장조절물질의 영향)

  • 전지혜;예병우;양미희;박재복
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.125-128
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    • 1997
  • The most effective cytokinin source for adventitious shoot regeneration of in vitro grown leaves from 'Gala' apple was BA with the concentration of 4.0 mg/L, while auxin source was IAA, IBA and NAA with the concentration of 0.1 mg/L, respectively. As the result of combinational treatment of BA and NAA, 6.0 or 8.0 mg/L BA with 0.5 mg/L NAA was effective for adventitious shoot regeneration from leaf tissues of 'Gala', 0.1 mg/L NAA + 8.0 mg/L BA and 1.0mg/L NAA + 8.0 mg/L BA for internode, and 0.1 mg/L NAA + 4.0 mg/L BA for petiole.

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A Study on acclimatization culture of the tissue cultured seedings in Zantedschia spp. (유색칼라 기내배양묘의 순화재배에 관한 연구)

  • Park, N.B.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.5 no.1
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    • pp.20-27
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    • 2003
  • This experiment was carried out to develop an acclimatization system of tissue cultured seedling in Zantedeschia spp. There was not different in percentage of leaf emergence by the media compost of acclimatization but tuber was enlarged in sand(50%)+peatmoss(50%) treatment after 6 months growth. The optimum time for the acclimatizantion of Zantedeschia spp. was from the 20th of March to 20th of April. During this time, rate of acclimatization reached over 90 percentage. However on the 20th of august, acclimatization was low to 64-70.4 percentage. Size of tuber was enlarged in planting on the 20th of March. The weight of tuber was better in 10×10cm density of planting than others.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Photomixotrophic Growth of Solanum tuberosum L. in vitro with Addition and Omission of Organic Materials at Thee Initial Sucrose Levels in the Medium (세 수준의 자당이 첨가된 배지에서 유기물의 첨가 유무에 따른 Solanum tuberosum L.의 기내 광혼합영양생장)

  • Jeong, Byoung-Ryong;Yang, Chan-Suk;Kim, Gyeong-Hee;Park, Young-Hoon;Kozai, Toyoki
    • Journal of Bio-Environment Control
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    • v.13 no.1
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    • pp.51-55
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    • 2004
  • The most commonly used inorganic nutrient compositions such as Murashige & Skoog medium have been optimized for heterotrophic growth. Therefore, they may not be optimal for photomixotrophic and photoautotrophic growth of plantlets. In photomixotrophic micropropagation, emdium sugar level is often lowered, while light and $CO_2$ levels in vessel are raised, and chlorophyllous explants are used to facilitate photosynthetic carbon acquisition. In a factorial experiment effect of addition (+) and omission(_) of organic materials (OM, 0.5 g ${\cdot}$ $m^{-3}$ each of thiamine, nicotinic acid and pyridoxine and 100 ${\cdot}$ $m^{-3}$ myo-inositiol) combined with three sucrose levels (0, 15, and 30 kg ${\cdot}$ $m^{-3}$) was tested on the growth of potato plantlets. Each of nodal cuttings with a leaf was cultured on 0.1${\times}$$10^{-4}m^{-3}$) MS agar ( 8 kg ${\cdot}$ $m^{-3}$) medium (pH 5.80 before autoclave) in glass test tubes (100 mm${\times}$25mm) capped with a sheet of transparent film with a 6 mm diameter gas permeable filter (5.1 air exchanges ${\cdot}$$h^{-1}$). Cultures were maintained in a room for 27 days at $23^{\circ}C$, 50% RH, 350-450${\mu}mol\;{\codt}\;mol^{-1}CO_2$, 16 h ${\cdot}$ $d^{-1}$ photoperiod at 13${\mu}mol\;{\codt}\;m\;{\codt}\;s^{-1}$ PPFD provided by white cool fluorescent lamps. Growth of potato plantlet in the +OM and -OM treatments were similar, while medium pH was 0.2 scale lower in the latter. Dry weight, % dry matter, and stem diameter enhanced, while shoot to root dry weight ratio, leaf area, chlorophyll concentration per gram dry weight, and medium pH decreased with increasing initial sucrose level. Interaction between OM and sucrose levels was observed in shoot length and medium pH. Results indicate that OM can be omitted from the medium without detrimental effect while addition of sucrose was beneficial for the photomixotrophic growth of potato plantlets under raised light and $CO_2$.

Development of In Vitro Seedling Screening Method for Selection of Resistant Rice Against Bakanae Disease (벼 키다리병 저항성 검정을 위한 기내 유묘 검정법 개발)

  • Lee, Yong-Hwan;Lee, Myeong-Ji;Choi, Hyo-Won;Kim, Sung-Taek;Park, Jin-Woo;Myung, Inn-Shik;Park, Kyung-Seok;Lee, Se-Weon
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.288-294
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    • 2011
  • Rice bakanae disease, caused by the fungus Fusarium fujikuroi, is one of the most important rice diseases and distributed widely in Asia. Resistance screening system in rice field had been established. However, the evaluation results of the system vary according to the environmental conditions when the test is conducted. To develop precise and rapid evaluation method of disease resistance of rice to bakanae disease, in vitro screening system was attempted in this study. The six cultivars namely, 'Nampyeongbyeo', 'Junambyeo', 'Chucheongbyeo', 'Samcheonbyeo', 'Odaebyeo' and 'Hwasinbyeo' were tested. They were planted onto MS agar medium (10 ml) in test tube ($450\;mm{\times}{\phi}30\;mm$) and incubated at $25^{\circ}C$ and $28^{\circ}C$ in growth chamber under 12 hr light condition. Symptoms of over growth appeared a few days after seeding and then seedling were withered 2-3 weeks after over growth. The disease symptoms such as leaf dryness on top of rice were appeared in the 'Nampyeongbyeo' from 28 days at the concentration of $10^5$ spores/ml culturing at $28^{\circ}C$ and then withered completely 35 days after seeding. Whereas the other varieties withered entirely 19-23 days after seeding. Using the in vitro seedling screening method, 72 rice varieties were investigated to select resistant cultivar. Finally, two resistant cultivars ('Nampyeongbyeo' and 'Inwolbyeo') and seven moderately resistant cultivars ('Hwadongbyeo', 'Seokjeongbyeo', 'Samgwangbyeo', 'Sampyeongbyeo', 'Nonghobyeo', 'Heukjinjubyeo' and, 'Joanbyeo') were selected. If in vitro seedling screening method was used for evaluation of bakanae disease resistance, it would be completed within 35 days after sowing of rice seed.