• Title/Summary/Keyword: MS medium

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Cotyledon Structure and Germinability of Somatic Embryos Formed from Inflorescence Explants of Cnidium officinale M. (천궁 (Cnidium officinale M.)의 화기절편으로부터 형성된 체세포배의 자엽구조와 발아)

  • 조덕이;이은경;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.137-142
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    • 2000
  • Calli were induced from the explants of infloresence, petiole and leaf blade of Cnidium officinale on MS medium with 2.4-D, while embryogenic callus was induced only from inflorescence explants. Somatic embryos of 78 per explant were formed during subculture of the explants on medium without 2.4-D after culture on medium with 2 mg/L 2.4-D. Cotyledonary variation, cup-shaped cotyledon of 49% and other abnormal cotyledons of 13.5 % was observed on the somatic embryos. However this variation could be overcomed by the addition of activated charcoal onto culture medium. Somatic embryos at cotyledonary stage germinated on MS basal medium but the germination rate was very poor, blow 50 %. Somatic embryos on the medium with activated charcoal showed improved germinaton.

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Somatic Embryogenesis and Plant Regeneration from Poinsettia (Euphorbia pulcherrima L.) Stem Explants (포인세티아 줄기조직배양에 의한 재분화체 제조.)

  • Hee-Sung Park
    • Journal of Life Science
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    • v.8 no.6
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    • pp.623-626
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    • 1998
  • Conditions for somatic embryogenesis and plant regeneration from stem tissues of Euphorbia pulcherrima were esta-blished. Explants from leaf, petiole, stem were examined for their embryogenesis on MS solid medium supplemented with plant growth hormones in combination at various concentrations. From leaf or petiole explants, callus was indu-ced well but never proceeded to the embryonic stage in our expermental conditions. From stem explants, however, multiple shoots following callus induction emerged in about 6 to 8 weeks on MS agar medium supplemented with 1.5 mg/L of benzyladenine. Upon transfer, roots were developed on hormone-free MS solid medium.

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In vitro plant regeneration from axillary buds of Hibiscus syriacus L.

  • Jeon, Seo-Bum;Kang, Seung-Won;Kim, Wan-Soon;Lee, Gung-Pyo;Kim, Sun-Hyung;Seo, Sang-Gyu
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.174-178
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    • 2009
  • Presently, we report a simple, reproducible and high frequency plant regeneration in Hibiscus syriacus L. using axillary buds. H. syriacus was regenerated from axillary buds directly or through a callus phase. Regenerated shoots were directly induced from young and fresh axillary buds cultured on Murashige and Skoog medium (MS) supplemented with 0.01 mg/L of the growth regulator thidiazuron (TDZ) after 2 weeks of culture. Directly induced shoots were transferred to hormone-free MS medium and root development was observed after 6 weeks. On the other hand, old and stale axillary buds were regenerated to shoots via callus induction on MS medium containing 0.01–2 mg/L TDZ after 4 weeks. A TDZ concentration of 0.01 mg/L was most effective in callus formation. Green callus was transferred to MS medium containing 0.01 mg/L α-naphthalene acetic acid (NAA) and 0.5 mg/L benzylaminopurine (BA). After 4 weeks, callus had developed into multiple shoots. Plantlets were formed from 10 week cultures of single shoots on hormone-free MS medium. Regenerated plantlets were cultured on MS medium for one month and then transferred to pots containing garden soil. Potted plants were acclimatized for one month and grown to maturity under greenhouse conditions. The present study has shown that various concentrations of plant growth regulator can be effective for in vitro plant regeneration of H. syriacus. The direct and indirect regeneration protocol presented here will be useful for understanding the manipulation and propagation of H. syriacus.

High Frequency of Callus Induction, its Proliferation and Somatic Embryogenesis in Cotton (Gossypium hirsutum L.)

  • Haq, Ikram-ul;Zafar, Yusuf
    • Journal of Plant Biotechnology
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    • v.6 no.1
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    • pp.55-61
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    • 2004
  • Callus induction and somatic embryogenesis are fundamental to cotton tissue culture biotechnology. An efficient protocol for callus induction, somatic embryogenesis and their maturation have been developed to regenerate plantlets from cotton (Gossypium hirsutum L.) variety coker 312. Embryogenic callus was initiated from hypo-cotyl region that was used as an explant at seedling stage when it was about 7-8 days old. Callus induction was achieved through culturing hypocotyls (5-7mm) on $MS_{1a} medium supplemented with 2,4-D (0.1 mg/L) and KT (0.5 mg/L) for six weeks. A friable, colorless, bulky and well proliferating callus becomes greenish with the addition of NAA (2.0 mg/L), ZT (0.1 mg/L) and removal of 2,4-D (M $S_{1b}$) cultured for two weeks then again transferred to $MS_{1a}. 2,4-dichlorophenoxyacetic acid (2,4-D) promoted the proliferation of embryogenic callus, but had a negative effect on the differentiation and germination of somatic embryos. ZT (0.1mg/L) and activated charcoal (2g/L), both hormones play an important role in differentiation and germination of somatic embryos in hypocotyls derived embryogenic callus but in case of cotton, such a capability have been observed on MS medium with 1.92 g/L $KNO_3$, but it is considered to attain somewhat more improvement. High embryogenesis frequency was achieved through nutrient deficient stress treatment. The frequency of globular embryogenesis (two-three folds) was achieved when well proliferating callus was (from $MS_{1a}$ media) cultured on MS (1/5 strength) medium for four weeks. Here the development of anthocyanins is the best indicator for somatic embryogenesis. However, when embryoid callus was cultured on MS (full strength) medium, the globular embryos were developed into normal plantlets immediately. In this procedure 27.49% cotyledenary embryos were developed. Of that 70% cotyledenary embryos were developed not only into normal plantlets but rooted simultaneously, when cultured on MS (with 0.05 mgg/L giberrelic acid) medium. So complete plants could be regenerated through somatic embryogenesis from hypocotyl explants within 6 months.s.

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 Propagation of Lilium Asiatic Hybrid 'Hae Hwa' via the Formation of Shoot Clusters (신초 Cluster 형성에 의한 Lilium Asiatic Hybrid 'Hae Hwa'의 기내번식)

  • Han, Bong-Hee;Yu, Hee-Ju;Yae, Byeoung-Woo;Goo, Dae-Hoe
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.19-23
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    • 2002
  • This experiment was conducted to micropropagate bulblets via shoot cluster formation and massproduce normal bulblets from the sections of proliferated shoot clusters in Lilium asiatic hybrid 'Hae Hwa'. The induction of shoot clusters from the culture of bulblet sections was more effective than that of bulb scales on MS medium with 1.0 mg/L BA and 0.5 mg/L IAA. Proliferation of shoot clusters from the formed shoot cluster sections was the most favorable on medium containing 5.0 mg/L BA and 0.5 mg/L IAA. The formation and the growth of bulblets from shoot cluster sections were achieved effectively on medium with 60∼90 g/L sucrose. The leaves derived from shoot clusters grew vigorously but the bulblets from shoot clusters grew very poor in 5L air-lift bioreactor culture. By the addition of 30 mL fresh liquid medium containing doulble strength MS salts, 250 g/L sucrose and 5 g/L activated charcoal after 8 weeks in the shoot cluster culture on MS medium with 5.0 mg/L BA and 0.5 mg/L IAA, the number of bulblets was increased in light condition, but the growth of bulblets was not affected by light. Bulblet production was possible with the bulblet product at 53 to 68 mg in fresh weight by liquid medium addition after the proliferation of shoot cluster.

Effects of Media, Hormones, and PFP on the Proliferation of Red Callus in Leaf Tissue Cultures of Garden Orach(Atriplex hortensis L.) (Garden Orach 조직배양에서 적색 캘러스의 증식에 미치는 배지, 호르몬 및 PFP의 영향)

  • 이주경
    • Korean Journal of Plant Resources
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    • v.7 no.2
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    • pp.171-175
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    • 1994
  • The effects of medium, hormones, and PFP on the proliferation of red callus in leaf tissue cultures of Garden orach(Atriplex hortensis L.) was investigated. As a result,88% of leaf tissues formed eallus on MS nledium containing 2mg/$\ell$ 2,4-D. Fresh weight of callus was higher on MS medium than on Bsand NN media. It was also found that 2, 4-D was more effective than Dicamba and Picloram. The op-timum concentrations of hormones for callus proliferation depended on culture media. Isolated red cal-lus grew markedly both on MS medium supplemented with 1-2mg/$\ell$ 2, 4-D and Bs medium contain-ing 2-4111g/$\ell$ 2,4D. Callus proliferated on B5 and NN media containing Dicabma Img/$\ell$ as well as onthe same media containing 2mg/$\ell$ Picioram. The addition of PFP concentrations of 2, 5, and 40mg/ $\ell$rcspectiely to culture medium caused increase of callus fresh weight, especially under light condition.

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Efficient Procedures for Direct Shoots Regeneration from Leaf Explants of Rehmania glutinosa Lib. (지황 잎조직 절편으로부터 신초 형성)

  • Hwang, Sung-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.6
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    • pp.273-277
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    • 2005
  • Adventitious shoots were directly induced from leaf explants of R. glutinosa, an important medicinal plant. Proliferating shoot cultures were obtained by culturing leaf discs on Murashige and Skoog(MS) medium alone or combination with auxins and cytokinins. MS medium supplemented with 1 $mg/{\ell}\;BA\;and\;2\;mg/{\ell}$ IAA was the most effective, providing high shoot bud formation frequency without formation of intervening callus. The effect of leaf age on adventitious shoot formation was also investigated. The maximum shoot bud production (93.4%) was achived using 3rd leaf from apex of 6 weeks old plantlets after seed germination. Plantlet were rooted on an half-strength MS (1/2MS) medium containing 0.1 $mg/{\ell}\;IBA$. This prtocol is useful for clonal propagation and Agrobacterium-mediated transforamtion in R. glutinosa.

Interaction Between time of Nodal Explant Collection and Growth Regulators Determines the Efficiency of Morus alba Micropropagation

  • Hassanein A.M.;Galal A.A.;Azooz M.M.
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.225-231
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    • 2003
  • The hormonal requirement suiting micropropagation of Morus alba during any season throughout the year was studied. Sprouting frequency from axillary buds of M. alba was greatly influenced by the time of explant collection, the highest value was achieved when nodal explants were collected at the end of bud dormancy period (late in March) and cultured on Murashige and Skoog (MS) medium supplemented with low concentration (0.5 mg/L) of BAP, kinetin or IBA (85-68%). In addition, they showed higher axillary bud sprouting on growth-regulators-free medium (49%) than others collected in autumn or winter and cultured on medium supplemented with various growth regulators (47-48%). Regardless of that period, young explants with greenish buds collected in summer exhibiting high sprouting frequency (66%) on MS medium supplemented with 0.5 mg/L kinetin and 0.5 mg/L GA3. Shoot multiplication via adventitious bud formation was achieved when the nodal explants were cultured on MS medium supplemented with 2 mg/L BAP and 0.2 mg/L IBA. Further multiplication via nodal explants of in vitro grown shoots was obtained on MS medium supplemented with 0.5 mglL BAP and 0.5 mg/L GA3. While half strength MS medium supplemented with low concentration (0.5 mg/L) of IBA, IAA or 2,4-D stimulated adventitious root formation, IBA was the best. After transfer the plantlets to the soil, acclimatization for three weeks was essential prerequisite for survival in high frequency (92%). Peroxidase activity is related to break of bud dormancy where maximum enzyme activity was detected when the lateral buds were induced to commence growth under field condition (early in spring) or in vitro.

Callus induction and high-efficiency plant regeneration via somatic embryogenesis in Papaver nudicaule L., an ornamental medicinal plant

  • Yang, Jing Li;Zhao, Bo;Seong, Eun-Soo;Kim, Myong-Jo;Kang, Won-Hee;Kim, Na-Young;Yu, Chang-Yeon;Li, Cheng Hao
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.261-267
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    • 2010
  • We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.