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Mass Propagation of Vitex negundo L., in vitro

  • Thiruvengadam, Muthu;Jayabalan, Narayanasamypillai
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.151-155
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    • 2000
  • Shoot proliferation was obtained from shoot tips and nodal explants of Vitex negundo L. on MS medium supplemented with either BAP or KIN (0.1-2.0 mg/L) alone or in combination with NAA (0.1 mg/L). The concentrations of cytokinins combined with NAA produced multiple shoots from shoot tips and nodal explants. The highest mean percentage (84.3$\pm$8.0) of shoot multiplication's were observed on nodal explants in the presence of BAP (1.5 mg/L) and NAA (0.1 mg/L) followed by shoot tips (65.0$\pm$5.0). The regenerated shootlets were rooted on MS basal medium IAA, IBA, NAA (0.1-1.5 mg/L). The maximum number of roots (51.0$\pm$2.6) was achieved on the medium containing IBA (1.0 mg/L) followed by other auxins (NAA, IAA). The regenerated plants were successfully transferred to a mixture of vermiculate and soil. About 95% of the plantlets survived when transferred to the field.

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Rapid Propagation through Tissue Culture of Cudrania tricuspidata, Medicinal Plant

  • Lee, Cheul-Ho;Min, Ji-Yun;Jung, Ha-Na;Kim, Kyu-Sick;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.315-318
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    • 2007
  • An effective rapid propagation method was established through in vitro cultures of the medicinal plant, Cudrania tricuspidata. In vitro plantlets were obtained from in vitro germinated seeds. The various levels of cytokinins (BAP, Kinetin and TDZ) were tested on multiple shoot formation from plantlets. BAP (1.0 mg/l) treatment induced highest number of multiple shoots. Single shoot cultures gave higher initial shoot numbers than 5 shoots per culture. Among the various culture media, the shoot elongation was optimal on 2 MS basal medium without growth regulators. The IAA (2.0 mg/l) treatment induced highest number of roots. IBA (2.0 mg/l) treatment more promoted in vitro root growth than other concentrations. Rooted shoots were transferred directly to small pots with an artificial soil and successfully acclimatized.

Induction of Callus and Culture of Protoplasts from Cotyledon-Originated Callus in Alnus hirsuta (물오리나무(Alnus hirsuta)의 칼루스 유도 및 자엽유래 칼루스로부터의 원형질체 배양)

  • 김형하
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.253-257
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    • 1990
  • Callus-inducing ability of Alnus hirsuta was examined by culturing various tissues (leaf, hypocotyl, cotyledon and seed) on NT (Nagata & Takebe) medium, supplemented with 2.5$\mu$M 2,4-D. Leaf-originated callus was cultured on media varying in auxin (IBA and NAA) and cytokinin (BAP) concentrations to examine the effects of auxin and cytokinin on callus growth. Maximum growth was obtained at 10 $\mu$M IBA+10$\mu$M BAP and 10$\mu$M NAA without cytokinin. Cell suspensions established from cotyledon-originated callus yielded viable protoplasts after incubation for 16-18 hours in an enzyme mixture (1% (w/v) Onozuka R-10 0.5% (w/v) Macerozyme, CPW salts and 13% (w/v) mannitol, pH 5.8). Protoplasts were cultured on NT medium, supplemented with glucose, hormones and coconut milk. After 6 weeks of culture, protoplasts sustained cell divisions to form microcallus, which showed various colors from red to white.

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In vitro propagation of Coleus forskohlii, an important medicinal plant

  • Yang, Deok-hun;Jeon, Manju-Meluttu-George;Jong-Seong
    • Plant Resources
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    • v.6 no.2
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    • pp.129-133
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    • 2003
  • For mass multiplication of an important medicinal plant Coleus forskohlii, a procedure for the high frequency regeneration of Coleus forskohlii has been developed using leaf explants via callus culture. Callus formation occurred in MS medium supplemented with 1-2 mg/L each of NAA and BAP. A large number of shoots were formed on MS + 1 mg/L BAP from 50-60 days old greenish calli. Rooting of healthy shoots occurred on 0.1-0.4 mg/L NAA. The protocol described could be useful in future for genetic manipulation of this plant species.

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Effects of Phytohormones on the Viability and Cell Wall Regeneraton of Tobacco Protoplasts (연초 원형질체의 생존율과 세포벽 재생에 미치는 식물생장조절물질의 효과)

  • 김용옥
    • Journal of Plant Biology
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    • v.31 no.2
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    • pp.121-130
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    • 1988
  • In order to clarify effects of phytohormones on the viability and the cell wall regeneration of protoplasts isolated from Nicotiana tobacum L. var. BY4, protoplasts isolated from mesophyll tissue were cultured on the Murashige-Skoog liquid media supplemented with auxin(2, 4-D, NAA, IAA) and/or cytokinin (kinetin, BAP, 2ip). Viability of protopplasts was higher in the culture medium containing auxin and cytokinin, especially in the combination of 2, 4-D and BAP. The effectual cell wall regeneration of protolasts was observed when theprotoplasts were cultrued on the medium supplemented with auxin alone, especially with IAA. Cell wall regernation started from 2-3 days after culture and was not detected at budding regions. When the protoplasts were cultured on the phytohormone-free medium, the viability of protoplasts dramatically decreased 4 days after culture.

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Silver nitrate and silver-thiosulphate mitigates callus and leaf abscission during Shisham clonal micro-propagation

  • Raturi, Manoj Kumar;Thakur, Ajay
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.173-178
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    • 2021
  • Basal callus formation and leaf abscission is a problem in clonal micropropagation. We have described an in vitro clonal propagation protocol of Dalbergia sissoo Roxb (shisham) 'FRI-14' in which AgNO3 played important role not only in mitigating problem of leaf abscission and basal callus, but also improved shoot induction and multiplication. Best induction and shoot multiplication was obtained on MS media with 1.5 mg/l 6-BAP and 10 mg/l AgNO3 and half-strength MS media with 0.5 mg/l 6-BAP, 2 mg/l AgNO3 and 50 mg/l Adenine sulphate whereas best ex vitro rooting was obtained with 200 mg/l IBA in pulse treatment.

Characterization of Albino Tobaccos (Nicotiana tabacum L.) Derived from Leaf Blade-Segments Cultured in vitro

  • Bae, Chang-Hyu;Tomoko Abe;Lee, Hyo-Yeon;Kim, Dong-Cheol;Min, Kyung-Soo;Park, Kwan-Sam;Tomoki Matsuyama;Takeshi Nakano;Shigeo Yoshida
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.101-107
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    • 1999
  • The leaf blade-segments of albino tobacco (Nicotiana tabacum L.) were cultured on MS media containing different concentrations of BAP (0, 0.4, 2.2, 4.4, 22.2 ${\mu}{\textrm}{m}$) with or without NAA (0, 0.5, 2.7 ${\mu}{\textrm}{m}$). Multiple shoots were induced on the media containing 0.4 to 2.2 ${\mu}{\textrm}{m}$ BAP. The best condition for multiple shoot induction with root formation was MS media containing 4.4 ${\mu}{\textrm}{m}$ BAP and 0.5 ${\mu}{\textrm}{m}$ NAA. The regenerated albino plants showed a significant reduction in accumulation of chlorophylls and carotenoids. The drastic reduction of the pigments content was associated with the distinct alterations in gene expression in the albino plants. firstly, the expression of plastid genes, such as rbcL, psbA, 165 rDNA and 235 rDNA, was reduced at the level of transcripts in the regenerated albino plants. Secondly, the alteration of structure of the plastid genes was not detected in the albino plants. However, the copy number of the plastid genes whose transcription level was reduced greatly was increased approximately two-fold, although the transcriptions of nuclear gene (255 rDNA) showed the wild-type level.

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Development of an efficient protocol for high-frequency regeneration system in Hibiscus syriacus L.

  • Seo, Sang-Gyu;Ryu, Sun-Hee;Zhou, Yang;Kim, Sun-Hyung
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.164-170
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    • 2017
  • A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.