• Title/Summary/Keyword: explant

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Effect of Cytokinin and Putrescine on Plant Regeneration from Leaf Explant of Rhodiola sachalinesis A. Bor (홍경천 (Rhodiola sachalinensis A. Bor) 잎절편으로부터 식물체 재분화에 미치는 Cytokinin과 Putresine의 영향)

  • Bae Ki-Hwa;Lim Soon;Yoon Eui-Soo;Shin Cha-Gyun;Kim Yoon-Young;Kim Yun-Soo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.195-199
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    • 2005
  • The effects of cytokinin and putrescine on adventitious shoot induction from leaf explant of Rhodiola sachalinensis A. Bor were investigated. Among cytokinin used in this experiment, BA was more effective on adventitious shoot induction and shoot elongation than kinetin. Especially, 1 mg/L BA was the best to increase adventitious shoot induction (71%) and shoot elongation (3.0 mm). In addition, 100 mM putrescine in MS medium with 1 mg/L BA was higher in adventitious shoot induction (93%) and shoot elongation (3.8 mm) than single treatment of 1 mg/L BA. Adventitious shoots induced in this experiment rooted on 1/2 MS medium and acclimated over 95% on composed soil (peatmoss:sand=1:1).

High-frequency shoot regeneration from leaf explants through organogenesis in bitter melon (Momordica charantia L.)

  • Thiruvengadam, Muthu;Rekha, K.T.;Yang, Chang-Hsien;Jayabalan, Narayanasamypillai;Chung, Ill-Min
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.321-328
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    • 2010
  • An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g $1^{-1}$ sucrose, 2.2 g $1^{-1}$ Gelrite, and 7.7 lM naphthalene acetic acid (NAA) with 2.2 ${\mu}M$ thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30-40 shoots per explant) was achieved on MS medium containing 5.5 ${\mu}M$ TDZ, 2.2 ${\mu}M$ NAA, and 3.3 ${\mu}M$ silver nitrate ($AgNO_3$). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 ${\mu}M$ gibberellic acid ($GA_3$). The elongated shoots were rooted in MS medium supplemented with 4.0 ${\mu}M$ indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.

Changes of RNA and Protein During Callus Induction and Plant Regeneration from Perilla frutescens (들깨로부터 Callus의 유기와 재분화에 따른 단백질 및 RNA의 변화)

  • 정상훈;양선경;김현경;정대수;조영수;김도훈
    • Journal of Life Science
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    • v.9 no.1
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    • pp.29-34
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    • 1999
  • Cotyledon and hypocotyl explants of perilla were cultured on MS medium containing a combined concentration of BA(0.5, 1.0 and 1.5mg/$\ell$) and NAA(0.1, 0.5 and 2.0mg/$\ell$) in order to regenerate the explant and induce the callus. The best regeneration of the explant and induction of the callus were observed in a combined concenteration of 0.5mg/$\ell$ of BA and 0.5mg/$\ell$ NAA both in cotyledon and hypocotyl explants. In cotyledon explants, rooting was achieved upon transferring shoots to MS medium containing 0.5mg/$\ell$ of BA and 0.1mg/$\ell$ of NAA. We also investigated the change of protein and RNA content on developmental stage of callus and plant regeneration of perilla. Protein content was increased but RNA content was decreased as the culture period increases. The banding pattern of polypeptide revealed that both 30KD and 45KD polypeptides were obvious in cotyledon obtained from pre-culture explants, but only 30KD polypeptide was further getting obvious as the culture period increases.

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In Vivo and In Vitro Rooting of Rehmannia glutinosa Plantlet Regenerated in Vitro (기내증식된 지황묘의 기내 및 기외 발근)

  • 백기엽;유광진;박상일
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.335-340
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    • 1997
  • 100% root formation in in vitro cultures was observed regardless of kind and levels of auxin used and explant source. The number of roots/explant was increased in 0.5~1.0 mg/L IAA treatment. Thicker roots were observed with the addition of 9% sucrose compared with medium containing lower sucrose concentrations. Paclobutrazol and chlormequat had no effect on tuberization of formed roots but slightly increased the number of root. In in vivo rooting, soaking of regenerated shoot cuttings to 100 mg/L IBA for 15 to 60 minutes was found effective. Treatment of 0.1% IBA rooting powder and planting in rooting medium composred of vermiculite(1) : perlite(1) gave 100% rooting and survival.

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Regeneration of Adventitious Shoots from Populus deltoides Bartr. (미류나무 엽육조직에서 식물체 재분화)

  • SUL, Ill-Whan;SHIN, Dong-ill
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.329-334
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    • 1997
  • Adventitious organs (shoots and roots) from mature leaves of Populus deltoides (Eastern cottonwood) clones were induced on WPM (Woody Plant Medium) containing fructose as a carborn source and different combinations of either BA (Benzyl aminopurin) or TDZ (Thidiazuron) with NAA (Naphthaleneacetic acid). The morphologies of adventitious shoots were different depending on the kinds of growth regulators and the length of shoots induced on the medium containing BA were generally longer than that of TDZ. BA 0.5 mg/L and TDZ 0.01 mg/L combined with either 0.05 or 0.1 mg/L NAA showed the highest values in the frequency of regeneration (63% to 100%) the number of shoots per explant and in the organogenic sites per explant among tested three clones (A, B and C). Although there was a minor clonal difference in the development of adventitious shoots from leaf segments, their responses to growth regulators were very similar.

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Effects of Growth Regulators and Sucrose Concentrations on the Bulblet Formation through In Vitro Culture of Scale Segment in Nerine bowdenii (네리네(Nerine bowdenii)의 기내 인편배양시 자구형성에 미치는 생장조절제와 Sucrose 농도의 영향)

  • Lee, Seung-Yeob;Ahn, Jeong-Ho;Park, Yun-Jum
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.139-143
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    • 2004
  • The twin-scale segments of nerine (Nerine bowdenii) were cultured to investigate the influence of NAA, BA and sucrose concentrations on in vitro bulblet formation. The formation of bulblets from twin-scale segments showed a good response both the percentage of bulblet formation and the number of bulblets per explant on MS medium supplemented with 1mg/L NAA and 2 mg/L BA. Formation of bulblet showed the highest efficiency on medium containing 30g/L, and the formation of bulblets was strongly inhibited on medium containing over 90g/L. When the twin-scale segments formed bulblets were subcultured three times to the same medium by 60 day subculture interval, the number of bulblets per explant was 6.5, 7.3 and 8.2 in order of first, second and third. The bulblets over 3mm in diameter were hypertrophied and rooted after transferring to the hormone-free MS medium. The plantlets over 50mm in height were successfully acclimatized in the soil mixed with the same volume of vermiculite and perlite, and the survival rate was over 95%.

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.

In Vitro Shoot Multiplication of Albizzia julibrissin Duraz. (자귀나무(Albizzia julibrissin Duraz.)의 기내 대량증식)

  • Ahn, Ji-Young;Kim, Jung-Hee;Kang, Ho-Duck
    • Korean Journal of Plant Resources
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    • v.19 no.4
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    • pp.524-529
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    • 2006
  • In vitro culture system was established to induce multiple shoots of Albizzia julibrissin Duraz. by investigating the effects of cytokinins. Cotyledon, hypocotyl and root explants were cultured on MS media supplemented with either three different plant growth regulators or their combinations. The most effective cytokinin sources were zeatin 2.0 + TDZ 0.5 mg/L in cotyledon, zeatin 1.0 mg/L in hypocotyl, and BA 0.2 + TDZ 0.01 mg/L in root explant for producing shoots ($5.67\;{\pm}\;1.20$, $19.50\;{\pm}\;3.50$, and $20.50\;{\pm}\;2.47$, respectively). Also, zeatin treatment was tended to induce more shoots rather than the combinations of other cytokinins. In addition, the root induced in 1/2 MS medium without any plant growth regulators was longer and thicker than treatments of IBA, NAA, IAA and 2.4-D as auxins. Overall, the highest average percent of in vitro shoot formation was 73% from three different types of explants with treatment of zeatin (1.0mg/L).

The effect of caspase-3 inhibition on interdigital tissue regression in explant cultures of developing mouse limbs

  • Kudelova, Judita;Tucker, Abigail S.;Dubska, Lenka;Chlastakova, Ivana;Doubek, Jaroslav;Matalova, Eva
    • Animal cells and systems
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    • v.16 no.4
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    • pp.295-301
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    • 2012
  • Interdigital tissue regression is one of the most well-known examples of embryonic programmed cell death, providing the mechanism behind separation of developing digits. Caspases have been shown to play a key part in this process, with activated caspase-3 localized between the developing digits. In caspase-3 knock-out adult mice, however, the digits are completely separated with no webbing. In other mutants with defects in the apoptotic machinery, such as Apaf1 deficient mice, interdigital tissue regression is initially inhibited but the webbing eventually disappears as alternative/additional cell death mechanisms step in. In order to investigate whether a similar temporal effect occurs after loss of caspase-3, we have used an in vitro approach to inhibit caspase-3 at specific times during digit separation. Previous limb explant culture approaches have encountered problems with proper limb development in culture, and thus a modified technique was used. The new approach enables detailed observation of the effects of caspase-3 inhibition on interdigital regression. Using these methods, we show that caspase-3 inhibition caused a delay in the loss of interdigital tissue compared with control explants, similar to that observed in Apaf1 mutant mice. Along with immunohistochemistry, active caspase-3 positive cells of the interdigital vs. digital regions were measured by flow cytometry. Notably, activated caspase-3 in vivo was found not only in the interdigital mesenchyme but also in the TUNEL negative digit region, supporting a role for caspase-3 in nonapoptotic events.

Auxin Affects on Production of Adventitious Roots and Secondary Metabolites in Echinacea angustifolia (오옥신 처리가 에키네시아의 부정근 및 생리활성물질 생산에 미치는 영향)

  • Jang, Young Se;Cui, Hai Yan;Lee, Eun Jung;Kim, Hae Won;Paek, Kee-Yoeup
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.479-486
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    • 2012
  • The production of adventitious roots derived from root explant of Echinacea angustifolia and its secondary metabolite content were assessed in different types and levels of auxin. The induction of adventitious roots from root explant cultured in Murashige and Skoog solid medium supplemented with 1.0 mg/L indole -3-butyric acid (IBA) attained highest as 20.87 mg fresh weight and 3.07 mg dry weight per culture but root suspension culture at the same concentration of IBA enhanced biomass production as 3.07 g fresh weight and 0.38 g per culture after 4 weeks in culture. 3.0 mg/L ${\alpha}$-naphthalene acetic acid (NAA) treatment had similar effect on root biomass production as 3.07 g fresh weight and 0.38 g per culture with liquid suspension culture, whereas adventitious roots exposed to over 3.0-5.0 mg/L IBA or 5.0 mg/L NAA were less responsive by reducing the number of adventitious roots and/or changing root morphology such as short and thick. The content of secondary metabolites such as phenolic, flavonoids and total caffeic acid in adventitious roots cultured on MS medium supplemented with 1.0 mg/L IBA were attained highest as 27.20, 9.60. 10.67 mg/g dry weight, respectively. Overall, the best production of root biomass and secondary metabolites were given by 1.0 mg/L IBA.