• Title/Summary/Keyword: multiple-shoot induction

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An Efficient In vitro Propagation of Zanthoxylum piperitum DC.

  • Hwang, Sung-Jin;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.4
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    • pp.316-320
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    • 2003
  • A protocol is described for rapid multiplication of Zanthoxylum piperitum DC. (Rutaceae), an important aromatic and medicinal plant, through shoot-tip explant cultures. Murashige and Skoog (MS) medium supplemented with various concentrations of N-6-benzyladenine (BA), N-6-benzylaminopurine (BAP) and thidiazuron (TDZ), in single or in combination with ${\alpha}-naphthaleneacetic$ acid (NAA), was used to determine the rate of shoot proliferation. N-6-benzyladenine (BA) used at 0.5mg/l, was the most effective in initiating multiple shoot proliferation at the rate of 23 microshoots per shoot-tip explants after 40 days of culture. Shoot multiplication increased 1.2-fold in each successive subculture. Induction of rooting (98%) was achieved by transferring the shoots to the same basal medium containing 2 mg/l indole-3-butyric acid (IBA). Plantlets went through a hardening phase in a controlled growth chamber, prior to in vivo transfer. These results represented that possible application for the mass production of plantlets through in vitro culture system of Zanthoxylum piperitum DC.

Improvement of Shoot Regeneration from Scutella-Derived Callus in Rice

  • Kim, Yong-Wook;Cho, Joon-Hyeong;Lee, Jang-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.52-60
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    • 2004
  • The optimized in vitro culture system was investigated for improvement of regeneration efficiencies by observing the responses of scutella-derived callus of Korean rice (Oryza sativa L.). Large variations of callus induction (43.9-93.9%) and shoot regeneration (0-88.7%) were observed among the rice cultivars depending on medium. However, shoot regeneration was significantly improved by selected utilization of basal medium, growth regulators, and carbon sources. N6 basal medium was more efficient for embryogenic callus induction than MS or LS basal medium, while MS was superior to N6 for shoot regeneration. The calli of highly regenerative cultivars grew faster and showed higher rates of green tissue formation (GT) and shoot regeneration (SR) and lower rate of callus browning (CB) than those of recalcitrant cultivars. Although a higher level of kinetin stimulated the GT and SR in highly regenerative cultivars, $10\textrm{mgL}^{-1}$ kinetin generally suppressed the GT and SR, while CB was accelerated compared to $2\textrm{mgL}^{-1}$ kinetin. Additional benefits of sorbitol combined with maltose (or sucrose) under $5\textrm{mgL}^{-1}$ kinetin were certainly confirmed on regeneration efficiencies compared to sucrose alone as carbon source and osmotic regulator. This combination showed high rate of GT and SR with multiple shoots while low rate of CB. With MSRK5SM-Pr medium ($5\textrm{mgL}^{-1}$ kinetin, 3% sorbitol, 2% maltose, $500\textrm{mgL}^{-1}$ proline), the regeneration efficiencies of total 17 out of 24 cultivars were practically improved 160% on average compared to MSRK2S ($2\textrm{mgL}^{-1}$ kinetin, 3% sucrose) control medium. Especially, the medium was most effective to the cultivars showing a medium level of regenerability such as Daesanbyeo and Dongjinbyeo and Suwon477, enhancing efficiencies more than 300-600% compared to MSRK2S medium.

Plant regeneration through multiple-shoot induction and ex vitro rooting in Vaccinium oldhamii Miq. (정금나무(Vaccinium oldhamii Miq.)의 다신초 유도 및 기외발근을 통한 식물체 재분화)

  • Yun, Ayoung;Kim, Tae Dong;Kim, Ji Ah;Lee, Na Nyum;Cheong, Eun Ju;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.82-89
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    • 2022
  • In vitro techniques were developed for propagating Vaccinium oldhamii using shoots with apical buds. Explants having an apical bud were cultured on Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, and 5.0 mg/L of each zeatin, thidiazuron, 6-benzylaminopurine (BA), and 6-(γ,γ-dimethylallylamino)purine (2-iP) in order to induce multiple shoots. Among the tested treatments, the 2.0 mg/L of 2-iP proved to be most suited for the multiplication and growth of shoots; the multiple shoot induction rate was 100.0%, the average number of shoots was 7.4 per explant, and the average shoot length was 51.7 mm. The in vitro elongated shoots were rooted on half-strength MS medium containing various concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA). However, overall callus overgrowth was observed in all treatments and resulted in necrosis and abnormal shoot growth in root formation. A low concentration (0.5 mg/L) of IBA was appropriate for normal root development and the in vitro rooting rate was 30%. Ex vitro treatments on root formation using various concentrations of IBA with Talc powder and two types of rooting substrates (Flexi-Plugs or Horticultural soil) were examined. The ex vitro rooting rate (80%) and length of roots (32.9 mm) were obtained when the cut ends of the shoots were treated with 1.0 mg/L IBA and cultivated in Horticultural soil for 2 months. These findings suggest that ex vitro rooting is the more effective method for improving root formation in Vaccinium oldhamii than in vitro rooting.

Multiple Shoot Induction on the New Cltivar, Saxifraga fortunei 'Greenstar' by Different Media and Plant Growth Rregulators (배지 및 생장조절제 종류를 이용한 바위떡풀 신품종 '그린스타' 의 다신초 유도)

  • Suh, Jong-Taek;Ryu, Seung-Yeol;Yoo, Dong-Lim;Nam, Chun-Woo;Hur, Youn-Young
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.2
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    • pp.83-86
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    • 2010
  • This study was conducted to develop in vitro propagation techniques of new cultivar, 'Greenstar', bred by Highland Agriculture Research Center. The multiple shoot induction and plant growth of in vitro plant were analyzed by MS media concentration (1/2 MS, 1 MS and 2 MS medium), plant growth regulators and its proper concentration; CPPU [Forchlorfenuron(N-(2-chloro-4-pridyl)-3-phenylurea) (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), thidiazuron [(TDZ), (0.01, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$), zeatin (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), and BA [6-benzylaminiopurine(BA), (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$)] in MS (3% sugar and 0.8% agar with pH 5.7) media. The highest number of induced shoots, leaves and roots were shown in 1/2 MS medium concentration. On the 1/2 MS medium, shoot numbers, shoot length, leaf numbers and root numbers were 11.0, 1.9 cm, 24.7, and 8.0, respectively. On the absence of CPPU in the 1/2 MS medium, shoot length and root numbers was greater than CPPU treatment, but the highest number of shoots was induced by the $2.0mg{\cdot}L^{-1}$ of CPPU concentration in 1/2MS medium. TDZ, zeatin, and BA treatments were not effective on the induction of multiple shoot in vitro culture. As a result, in vitro culture of new Saxifraga fortunei, 'Greenstar' with $2.0mg{\cdot}L^{-1}$ of CPPU in 1/2 MS medium was most effective for the rapid multiplication.

Microprogation And Environment Conditions Affecting On Growth Of In Vitro And Ex Vitro Of A. Formosanus Hay

  • Ket, Nguyen-Van;Paek, Kee-Yoeup
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.6-7
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    • 2002
  • The goal of this research was to develop the effectiveness of in vitro culture method for A. formosanus and study the environment in vitro conditions affecting on growth. The first series of experiments were examined to investigate the response of three different basal media, MS (Murashige and Skoog, 1962), Knudson (KC; Knudson, 1946) and modified hyponex on growth and multiplication during in vitro culture. Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with BA (1 mg1$^{-1}$) or TDZ (1-2 mg1$^{-1}$). Addition of activated charcoal (1%) to the TDZ containing medium promoted rapid shoot tip proliferation (11.1 shoots per explant) but the same medium had an opposite effect resulting in poor proliferation in the nodal explants. However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 media supplemented with 2% sucrose and 0.5% activated charcoal. Using bioreactor culture for scaling up was also shown the best way for multiple shoot induction and growth of this plant.(중략)

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Studies on Multiplication of Cornus of officinalis by in vitro Culture I. Callus Induction, Shoot Propagation and Root Differentiation through Bud Culture (산수유(山茱萸)(Cornus officinalisis)의 기내증식(器內增殖)에 관한 연구(硏究) I. 액아배양(腋芽培養)에 의한 Callus 유기(誘起), Shoot 증식(增殖) 및 뿌리 분화(分化))

  • Park, Chung-Heon;Seong, Nak-Sul;Lee, Seung-Tack;Youn, Kyu-Bok;Son, Su-Gyu
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.1
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    • pp.63-69
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    • 1993
  • Present experiment were attempted to examine in vitro multiplication throughbud culture of Cornus officinalis. Bud derived shoot formation was established successfully on Murashige and Skoog's medium supplemented with $0.5mg\;/\;{\ell}$ BAP(N-benzyl amino purine). The shoot proliferation increased on the Driver Kuniyuki Walnut medium containing $0.5mg\;/\;{\ell}$ NAA(Napthalene acetic acid) and $0.5mg\;/\;{\ell}$ BAP. Addition of 2,4-D(2,4-Dichlorophenoxy acetic acid) to the media produced excessive callus inducton. IAA(Indole-3-acetic acid) and IBA (Indole-3-bu-tyric acid) enhanced multple shooting, and NAA showed callus induction and multiple shooting. Shoot growth was enhanced supplemented with 3% sucrose, $2g\;/\;{\ell}$ activated charcoal, and 1 / 4MS in organic salts. However, root formation of proliferated shoots was low about 5%

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Multiple Shoot Formation of Gentiana axillariflora Leveille by in Vitro Culture (큰용담의 기내증식에서 multiple shoot의 유기)

  • Lim, Jung-Dae;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.1
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    • pp.41-48
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    • 2000
  • This study was aimed to proliferate Gentiana axillariflora Leveille which is one of the important medicinal and ornamental plants, by establishment of multiple shoot formation and embryogenesis through tissue culture technique. Callus was formed on MS (Murashige and Skoog) medium supplemented with 2,4-D, CPA, but not formed with BAP. The addition of 2,4-D 2 mg/ l into the medium was effective for callus formation and the rate of callus formation was about 90%. Somatic embryos were obtained on MS medium for two months. When callus was cultured on MS medium with combination treatment of 2,4-D 0.5 mg/ l and BAP 0.5 mg/ l, the number of embryo formed was better than that of other single or combination treatments and the total numbers of embryo a were 18.8 (number of total embryo/number of explants incubated = 753/40) at mean. Callus induction from stem and node explants was increased by addition of TDZ 2 mg/ l in the presence of 2,4-D 2 mg/ l, respectively. The best result about the differentiation of shoots was obtained on MS medium added BAP 2 mg/ l from node culture. Multiple shoots from shoot apex were induced on MS medium containing BAP 1 mg/ I and TDZ 1 mg/ l , BAP 2 mg/ l and TDZ 1 mg/ l. The number of multiple shoots per one explant was above seventy plants. It was the most effective regeneration system for rapid multiplication of Gentiana axillariflora Leveille.

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Propagation of Bell Pepper (Capsicum annuum var. grossum) through tissue culture

  • Senarath, WTPSK;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.131-132
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    • 2003
  • Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

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Effects of BA and NAA on Adventitious Shoot Formation from Mature Zygotic Embryos of Stewartia koreana Nakai (노각나무 성숙 접합자 배로부터 부정아 유도에 미치는 BA 및 NAA 효과)

  • 손석규;조윤진;문흥규
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.272-277
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    • 2004
  • Zygotic embryos of Stewartia korean Nakai were cultured to determine the effects both of BA and NAA on adventitious shoot induction. Multiple shoots (about 11 per explant) were formed when the embryos were treated with 1.0 mg/L BA alone. On the other hand, NAA appeared to inhibit shoot induction when treated with BA. Adventitious shoots looked differ in form and color by the combinations of BA and NAA treated. When both BA and NAA were present, the shoots became short and developed dark color. The highest rooting was observed at 0.5mg/L NAA. The results could be useful for the establishment of in vitro regeneration system for Stewartia koreana Nakai.

Soil Acclimatization of Calanthe discolor through Multiple Shoot Formation from Tissue Culture (새우난초(Calanthe discolor)의 조직배양으로부터 다신초형성을 통한 토양순화)

  • Bae, Kee-Hwa;Yoon, Eui-Soo;Yun, Pil-Yong;Choi, Yong-Eui
    • Korean Journal of Plant Resources
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    • v.23 no.1
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    • pp.7-13
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    • 2010
  • This experiment was conducted to establish the micropropagation of Calanthe discolor through multiple shoot formation from the culture of leaf, corm and root explants. Frequency of adventitious shoot formation from leaf explants was higher than those of corms and root explants. Frequency of adventitious shoot formation on medium with various concentrations of BA (0. 1.0, 3.0, and 5.0 mg/L) and NAA (0, 0.1, 0.5, and 1.0 mg/L) was tested. The maximun induction of adventitious shoot was obtained on half strength Murashige and Skoog (MS) medium supplemented with 3.0 mg/L BA and 1.0 mg/L NAA after 6 weeks of culture. Multiple shoots were transferred onto half strength MS medium with various concentrations of GA3 (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L). The number and length of multiple shoots on medium were highest on medium with 3.0 mg/L GA3. All the adventitious shoot grew well and rooted on half strength MS medium with 3.0 mg/L NAA. The plantlets were acclimatized up to 100% on sand with TKS-II or pearlite with TKS-II.