• Title/Summary/Keyword: in vitro bud culture

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In vitro seed germination and callus formation on flower bud of Korean mistletoe ( Viscum album L. var. cololatum [Kom.] Ohwi) (겨우살이 종자 발아 및 화아 배양에 의한 캘러스 형성)

  • Kim, Suk-Weon;Ko, Suk-Min;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.47-53
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    • 2008
  • Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.

Studies on the Differentiation of Chondrogenic Cells in Developing Chick Embryo I. Cellular Aggregation and Chondrogenesis (발생계배 연골세포의 분화기구에 대한 연구 I. 세포응집과 분화와의 관계)

  • 박대규;손종경;유정아;유병제;강신성
    • The Korean Journal of Zoology
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    • v.33 no.3
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    • pp.310-321
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    • 1990
  • To establish the in vitro culture system and quantitation for chondrogenesis, and to investigate the relationship between cell aggregation and chondrogenesis, chick limb bud mesenchymal cells of Hamburger-Hamilton stage 23/24 were micromass cultured in various cell densities. The chondrogenesis was assayed based on checking the alcian blue-stained nodule numbers, the amount of alcian blue extraded, the change in cell numbers, the rate of [35 S] sulfate incorporation and expression of type II collagen. Mesenchymal cells plated with an initial density of high (1 x 107 cells/ml)- and intermediates (5. $\times$ 106 cells/ml)-density were differentiated into cartilage. On the other hand, the cells of low density (2 x 106 cells/mi, 5 $\times$ 105 cells/ml) of stage 23/24 cells and the stage 18/19 cells in three kinds of cell density did not differentiate into cartilage even though the cells formed an aggregated core at the center of cultured mass. From these results and others obtained in this study, it can be stated that the stage 23/24 mesenchymal cells are likely to pass over the aggregation step and have the potentiality to differentiate into chondrocytes. Thus chondrogenesis in vitro can be observed when mesenchymal cells are plated over the threshold density of 5 $\times$ 106 cells/ml. Hyaluronidase (HAase) activity was relatively constant throughout the culture, suggesting that the role of HAase may not be important for the cells of stage 23/24.

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Rapid Micropropagation by Axillary Buds Cultures of Smilax china

  • Song, Hyun-Jin;Sim, Seon-Jeong;Jeong, Mi-Jin;Heo, Chang-Mi;Kim, Hak-Gon;Jeong, Gwon-Yong;Heo, Su-Yeoung;Choi, Yong-Weon;Park, Geun-Hye;Yang, Jae-Kyung;Moon, Hyun-Shik;Choi, Myung-Suk
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.39-44
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    • 2010
  • An efficient method for the rapid propagation of Smilax china from axillary buds was established. Plants with thick leafage were selected from Korea native S. china population. Axillary buds of S. china collected from selected plant and were cultured in various culture media (2MS, MS, 1/2MS, WPM, B5 and SH medium). Shoot was induced from axillary bud on MS basal medium after 4 weeks of culture. 1/2MS medium showed a higher growth rate than those of the others, while the lowest shoot growth was obtained in 2MS medium. Among the sucrose concentrations, 5% sucrose was the optimum level for shoots growth from axillay buds. Among cytokinins, $0.5mgL^{-1}$ 6-benzylaminopurine (BAP) treatment showed the best performance on shoot multiplication, yielding average shoot multiplication forming about 2.4. Rooting was induced directly near the base of the shoot on 1/2MS medium containing with three-auxins ${\alpha}-napthalene$ acetic acid (NAA), indole acetic acid (IAA) and ${\beta}-indolebutyric$ acid (IBA) (0.5 and $1.0mgL^{-1}$). The $1.0mgL^{-1}$ IBA treatments induced earliest rooting with maximum of root number and root growth. These rooted plantlets were successfully transferred to pots for 4 weeks hardening process, and were transferred to soil with above 90% survival rate.

In vitro mass propagation and acclimatization of Haworthia truncata (하월시아 옥선(Haworthia truncata)의 기내 대량 증식 및 순화 조건 구명)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.127-135
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    • 2019
  • The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA $1{\sim}2mgL^{-1}$ treatment, and NAA $1mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment. The flower stalk showed 75% callus formation rate, at NAA $2mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA $0.1mgL^{-1}$ treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA $1mgL^{-1}$ + BA $0.1mgL^{-1}$ treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

Microtuberization and Morphological Development by Culture Condition In Vitro Node Culture of Potato (감자절간 기내배양에서 소괴경의 형성과 형태적 발달)

  • Hwang, Hye-Yeon;Lee, Young-Bok
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.331-338
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    • 2007
  • One-node stem pieces ca. 1 cm in length containing a axillary bud and a fully expanded leaf were obtained from it in vitro plants of potato (Solanum tuberosum L.). Leaves were removed and the nodes were cultured on the MS medium to investigate the effects of temperature, day length, sucrose, and CCC in microtuber formation and development. The fresh weight of microtubers after 80 days increased significantly at 8% sucrose and $20^{\circ}C$ compared with $28^{\circ}C$. The tuberization and development were reduced at $28^{\circ}C$ except short-day treatment of 8 hours at 8% sucrose. The fresh weight and diameter were increased on the culture medium added CCC 500 mg/L. The potato tuberization was promoted under short daylength, and it showed great effect by treatment with the CCC. Though the tuberization was promoted at low temperature of $20^{\circ}C$ in a histologic change of an axillary bud part cell of a potato, the cells were able to observe the swelling growth. Swelling growth of tissue was stimulated in the darkness and was more remarkable by addition of CCC. In particular, in the visual ratio of cell division for each position in the tissue, the cortex part showed larger ratio of cell expansion than that of the pith part. The effect of CCC was identified at 8% sucrose in the darkness. The effect of CCC was not showed in sucrose 3% under long daylength of 16 hours. As a result, the fact of a substance with AGPase important for starch composition was certified by the result with the inclose of AGPase activity on high concentration of sucrose, CCC, and dark treatment by which tuber formation and development are promoted.

Micropropagation of Juvenile and Mature Tree of Corylopsis coreana by Axillary Bud Culture (액아배양에 의한 유묘 및 성숙 히어리나무의 기내번식)

  • Moon, Heung-Kyu;Noh, Eun-Woon;Ha, Yoo-Mi;Shim, Kyung-Ku
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.117-121
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    • 2002
  • We have developed an in vitro micropropagation system via shoot formation from axillary buds using nodal segments of Corylopsis coreana. Explants from both juvenile tree (one-year-old greenhouse stock seedlings) and mature tree (ten-years-old tree in nursery) were compared with regard to propagation efficiency. Combined treatment of both BA and zeatin were effective on shoot proliferation since the best result was obtained on MS medium supplemented with 0.5∼3.0 mg/L zeatin and 0.2 mg/L BA. Generally, juvenile explants were better in both shoot proliferation and growth than mature explants. However, as the duration of in vitro culture was proceed to 6 months, explants from mature tree also produced three shoots per explant. Distinctive differences in rooting and adaptability to soil of shoots obtained from mother trees. Whereas shoots originated from juvenile explants rooted as high as 97%, those from adult explants showed 62% rooting. Similar result was also observed in soil acclimatization. The plantlets derived from juvenile plants survived 67%, while only 48% of those from adult trees survived. The results showed a possibility of the micropropagation of Corylopsis coreana through shoot formation from axillary buds. In addition, the advance of the research still remain to enhance the frequency of acclimatization of plantlets from mature trees for practical application.

In vitro Plantlet Regeneration of Loblolly Pine, Pitch Pine, and Their Hybrid -The Culture of Embryonic Tissues- (조직배양(組織培養)에 의한 테다, 리기다 및 교잡종(交雜種) 소나무의 식물체(植物體) 번식(繁殖) -배조직(胚組織)의 배양(培養)-)

  • Yi, Jae-Seon
    • Journal of Korean Society of Forest Science
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    • v.78 no.4
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    • pp.401-411
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    • 1989
  • The embryos of Pinus taeda, P. rigida, and P. taeda ${\times}$ rigida were cultured for adventitious shoot regeneration in vitro. Culture media were modified from Gresshoff and Doy (MGD), Murashige and Skoog (MMS), Lloyd and McCown (MLM), and Schenk and Hildebrandt (MSH). NAA was added to initiation media at a concentration of 0.1 or 0.01 mg/l. BAP was used at the concentrations of 0.1. 0.5, 1, 2, or 5mg/l. Each explant was induced for 3-4 weeks on solid medium. All explants were cultured up to 16 weeks. Illumination was about $1506{\pm}540lux$ at the level of the tissues in the growth room with a temperature of $25{\pm}2^{\circ}C$. A 16-hour photoperiod per 24 hours was used. Half-strength medium was used for all the subcultures. For shoot production by loblolly pine, MMS, MLM, or MSH is preferred with 5 mg/l BAP with either 0.1 or 0.01 mg/l NAA. For shoot production by pitch pine, MMS, MLM, or MSH is recommended with 2 or 5 mg/l BAP with 0.1 mg/l NAA. For shoot production by the hybrid pine, MMS or MLM is more effective with 1, 2 or 5 mg/l BAP with 0.1 mg/l NAA. There were no differences recognized among the species tried in the patterns of bud formation and shoot development. Different composition of media, in major and minor salts or possibly in vitamins, should be tested for the two developmental stages of adventitious shoots ; the induction of shoot buds and the elongation of them into shoots.

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Somatic Embryogenesis from Various Parts of Muscari comosum var. plumosum

  • Xudong He;Ko Jeong-Ae;Choi Jeong-Ran;Kim Hyung-Moo;Kim Myung-Jun;Choi So-Ra;Kim Young-Gon;Kim Dong-Hee;Kim Hyun-Soon
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.427-431
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    • 2006
  • In vitro high-frequency plant regeneration of Muscari comosum var. plumosum through somatic embryogenesis was obtained via two developmental pathways: direct embryos and multiple shoots regenerated from embryogenic callus. Flower bud with pedicel, receptacle, petal and ovary wall, floral stalk and leaf as explants were cultured in MS medium supplemented with various plant growth regulators. Embryos formed directly from pedicel, receptacle and floral stalk. Depending on explant sources, the optimal medium was MS medium supplemented with 0.2 mg/L IBA and 0.3 mg/L BA, 3.0 mg/L IBA and 3.0 mg/L BA, and MS-free medium for pedicel, receptacle, and floral stalk, respectively. Multiple shoots regenerated from embryogenic cal]i which was initiated from petal, ovary and leaf were observed in MS medium with different concentrations and combinations of hormone. The most suitable medium for each type of explant was 3.0 mg/L IBA and 3.0 mg/L BA(petal and ovary) and 5.0 mg/L IBA and 5.0 mg/L BA (leaf) Furthermore, the combination of 0.1 mg/L 2,4-D and 1.0 mg/L BA was also good for all sources of explants not only for direct embryo formation, but also, for embryogenic callus induction.

Effect of Plant Growth Regulators on Bulblet Formation and Plant Regeneration in Fritillaria thunbergii Miq. (패모조직배양에서 생장조절 물질이 자구형성 및 식물체 재생에 미치는 영향)

  • Park, Chul-Hyoung;Ryu, Jeom-Ho;Han, Kwang-Soo;Doo, Hong-Soo;Choi, Sun-Young
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.2
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    • pp.119-125
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    • 1996
  • To improve the efficiency of mass propagation in vitro of Fritillaria thunbergii, bulb scale and nodes were cultured in LS medium supplemented with the combination of 2, 4-D and kinetin or NAA and BA. The number and size of bulb, the number of adventitious shoot, the ratio of callus formation, rooting, and the effects of light and dark on the culture, plant regeneration from calli, and the gelling substances were investigated. The combination of 2, 4-D and kinetin in media was more effective than the media of NAA and BA for the bulblet formation. The media supplemented with 2 mg/L 2, 4-D and 1 mg/L kinetin, $1{\sim}2\;mg/L$ 2, 4-D without kinetin and $1{\sim}3\;mg/L$ BAA only were effective in the adventitious shoot development. Callus formation and root formation, respectively were effective in the medium supplemented with 2mg/L 2, 4-D and 1mg/L kinetin. In bulb formation, the medium with 5 mg/L kinetin was effective, and the most of bulbs were formed from the axillary bud of node part. In bulb formation, shoot growth, callus and root formation, the light culture for 16 hours per day was better than that in the dark culture. Bulb was nicely formed in the medium with 0. 2 mg/L 2, 4-D, 1 mg/L kinetin. The medium without hormone was most effective for plant regeneration. The phytagel was more effective than agar in the medium as the gelling agent.

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In vitro Variant Induction and Its Content of Gentiopicroside of Gentiana scabra BUNGE (용담(龍膽)의 기내변이주(器內變異株) 유도(誘導)와 변이주(變異株)의 Gentiopicroside 함량(含量))

  • Seong, Nak-Sul;Park, Chung-Heon;Kim, Kwan-Su;Lee, Seoung-Tack;Chang, Yeong- Hee
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.40-44
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    • 1995
  • This study was aimed to mutant induction in Gentiana scabra BUNGE through the tissue culture techniques and comparison of gentiopicroside content in each mutant plants derived from tissue culture. In order to induce the mutants of Gentiam scabra, seed, apical and lateral bud were soaked in $NaN_3$, EMS and MNU solutions and cultured on MS basal medium containing 0.5mg/l NAA, BA $NaN_3$, MNU-treated buds survived about 50% but not survived in EMS treatment. Seed germination rate was extremely low in EMS and MNU treatments but various types of mutant plantlets were obtained by those treatments. 63% of elongated type and 37% of dwarf type were obtained from tissue culture treated by various mutagen. Gentiopicroside contents of regenerated plantlets was analyzed. Root of dwarf type contained more gentiopicroside(1.383%) than that of elongation type (0.083%).

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