• Title/Summary/Keyword: explant

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Shoot Organogenesis and Plantlet Regeneration from Stem Explants of Cleome rosea Vahl (Capparaceae)

  • Claudia Simoes;Alessandra S. Santos;Norma Albarello;Solange Faria Lua Figueiredo
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.199-204
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    • 2004
  • The medicinal value of the genus Cleome justifies bio-technological studies of Cleome rosea, a Brazilian annual species from sandy coastal ecosystems (restinga), which have been submitted to an intense process of antropogenic degradation. In the present work, was analyzed the influence of cytokinins, 6-benzyladenine (BA) and 6-furfurylaminopurine (kinetin) added to the Murashige and Skoog medium (MS), on the proliferation capacity of explants from the stem axis (hypocotyl, node and internode) for a period of five monthly subcultures (150 days). Regardless of the explant sources, plantlet regeneration by direct and indirect organogenesis was observed. The largest number of shoots proliferated through direct organogenesis was obtained on medium with 4.4 $\mu{M}$ BA. Also, the highest proliferation capacity through indirect organogenesis was found on medium with 4.4 $\mu{M}$ BA + 4.6 $\mu{M}$ kinetin. The presence of kinetin alone was not effective for multiplication of the species. Elongation and rooting were obtained when shoots were transferred onto growth regulator-free medium, and acclimatization rates from 70% to 81% were achieved depending on explant sources used. Plants were then successfully established in soil and showed normal phenotypes.

Optimization of Regeneration Condition Under Agrobacterium-mediated Transformation in in vitro Cultured Korean Soybean

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.36-36
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    • 2018
  • Soybean is the important crop in Asian countries as protein source, oil production and animal feed. Improving soybean using genetic transformation is the principal tool in nowadays. Developing herbicide resistant transgenic soybean plants through Agrobacterium-mediated transformation has been worked in many previous studied. However, the transformation efficiency is still low. Many attempts try to find the optimum media condition for plant regeneration after infection. After transformation, the plant regeneration is very important condition to promote growth of transgenic plant. In this study, we optimized a regeneration condition for two Korean soybean cultivar, Dawonkong and Pungsannamulkong using cotyledon, cotyledonary nodes and hypocotyl as explant. The results showed that shoot regeneration of cotyledonary nodes on B5 medium containing 2 mg/L 6-benzylaminopurine showed the highest percentage of regeneration in Dawonkong (75.8%) while Pungsannamulkong presented high number of shoots 2.12 shoots per explant. For transformation condition, co-cultivation in 7 days showed a high number of GUS positive expression. Most of explants can survived under media including 5 mg/L of glufocinate which refers phosphinotricin for 2-week selection. Washing with 400 mg/L of cefotaxime in several times and selection in plant regeneration media with 400 mg/L of cefotaxime can prevent bacteria growth, effectively.

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Effect of Some Factors Affecting Callus Formation of Red Clover (레드 클로버의 캘러스 유도에 미치는 몇가지 요인의 영향)

  • Dae Young Son;Jin Ki Jo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.15 no.2
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    • pp.101-105
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    • 1995
  • This experiment was conducted to determine factors affecting callus formation of some varieties of red clover. Growth regulators. basal medium, medium pH, agar concentration, and plant age for explant were investigated. The result obtained were as follows. As auxin source, 0.1 mg/ l picloram was found to be effective on callus formation. Callus formation was the highest on PC medium among several growth media. The callus produced on PC medium was more friable than those produced on the other media. The medium pH 5.8 gave the best response for callus formation. The effective seedling age for callus formatiom was around 5-7 days old. In agar concentration, 0.5-0.8%(WN) was suitable for callus formation.

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A Tetraploid Induction in Hypericum patulum Thunberg by Colchicine Soaking Treatment (콜히친 침지처리에 의한 '망종화'의 4배체 식물유도)

  • Kwon, Soo Jeong;Cho, Kab Yeon;Kim, Hag Hyun
    • Korean Journal of Plant Resources
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    • v.26 no.2
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    • pp.284-288
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    • 2013
  • This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.

Effect of LEDs on shoot multiplication and rooting of rare plant Abeliophyllum distichum Nakai (희귀 수종 미선나무(Abeliophyllum distichum Nakai.)의 기내 증식 및 발근에 미치는 LED (light emitting diode) 효과)

  • Lee, Na Nyum;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.94-99
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    • 2014
  • This study was conducted to elucidate the effect of light sources and explant types on in vitro shoot multiplication and rooting of a rare and endangered plant Abeliophyllum distichum. Both apical buds and axillary buds were used as explants under 4 different light sources, cool white florescent light (F), 100% blue light-emitting diode (LED) (B), 50% blue and 50% red LED mixture (BR), and 100% red LED (R). Clear difference was observed in terms of shoot proliferation by light sources types but not by position-dependent explant types. Multiple shoot induction rates were enhanced under both B and BR light sources. Spontaneous rooting was induced in shoot induction medium under B light source. Both the rates of rooting and numbers of roots per explant were higher in apical bud explants compared to axillary bud explants. Interestingly R light source stimulated shoot elongation but inhibited root development. Therefore, our results suggest that the use of apical bud explants under B or BR light sources is suitable for in vitro micropropagation of a rare and endangered plant species, Abeliophyllum distichum.

Effect of explant parts and medium components on in vitro regeneration in Osmunda cinnamomea var. forkiensis (꿩고비(Osmunda cinnamomea var. forkiensis) 기내 포자체 재생에 영향을 미치는 배양부위와 배지구성물질)

  • Kwon, Hyuk Joon;Shin, So Lim;Lee, Cheol Hee;Kim, Soo-Young
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.448-453
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    • 2017
  • This study was carried out to find culture materials (explant parts) and medium components (medium type, sucrose and $NaH_2PO_4$ concentration) for in vitro propagation of Osmunda cinnamomea var. forkiensis sporophyte. The results of study: chopped segments of leaf blades, stipes, rhizomes and roots were cultured on a 1/2MS medium supplemented with 0.1% activated charcoal. Among these explant types, only the rhizome segments produced young sporophyte, regenerating vigorously on a 1/8MS medium. Adjusting the sucrose concentration to 2% and supplement to $50mg{\cdot}L^{-1}\;NaH_2PO_4$ in the 1/8MS medium proved to be more efficient for plant regeneration. Consequently, the addition of 0.1% activated charcoal to a modified 1/8MS medium (2% sucrose, $50mg{\cdot}L^{-1}\;NaH_2PO_4$, pH 5.8 and 0.8% agar) yielded the highest sporophyte regeneration.

Effect of Plant Growth Regulators, Media and Celling Agents on In Vitro Microtuber Production of Pinellia ternata Breit (1식물생장조절제, 배지와 고형지지물이 반하의 기내 소괴경 생산에 미치는 영향)

  • Kim, Yong-Kyung;Lee, Ji-Yeon;Kim, Eung-Hwi;Cho, Dong-Ha;Park, Sang-Un
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.3
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    • pp.210-214
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    • 2007
  • The study was carried out to establish in vitro microtuber production from leaf and petiole explant cultures of Pinellia ternata. Culture conditions were optimized by investigating the effect of plant growth regulators, different media, and gelling agents on the efficiency of microtuber indurtion. Among the different combinations of plant growth regulators tested, the combination of 0.1 mg/l 2,4-Dichlorophenoxyacetic (2,4-D) and 0.5 mg/l 6-benzylaminopurine (BAP) showed the highest yield fer microtuber production from leaf (3.9) and petiole (4.7) explant culture on MS medium far 6 weeks. SH(Schenk and Hildebrandt) medium was the most effective medium for microtuber induction and the half strength SH medium was better than SH medium for microtuber production from both leaf and petiole culture. Gelrite was better than agar in the formation of microtubers and 4% Gelrite showed the highest number of microtubers per explant frome leaf (5.9) and petiole (7.8) culture. Germination rate of microtubers after cold storaged for one months long was 86% from in vitro culture and 43% from autoclaved soil.

Micropropagation through Stem, Node-bud Shoot Tip and Bulblet Scale Culture in Fritillaria thunbergii Miq. (패모의 줄기, 마디, 정단 및 자구인편 배양에 의한 기내 증식)

  • Peak, Kee-Yoeup;Yu, Kwang-Jin;Seong, Nak-Sul;Choi, In-Sick;Cho, Jin-Tae
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.2
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    • pp.154-161
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    • 1994
  • This experiment was carried out to establish micropropagation system in Fritillaria thunbergii Miq. Through the culture of bulblet scales, stems, node-buds and shoot tips with special reference to the effect of physiological age of explant and plant growth regulators on bulblet formation. Number of formed bulblets was significantly increased in node-bud or stem tissue compared to scals segments and on the medium supplemented with kinetin than BA containing medium. Optimum levels of kinetin for bulblet formation from node-bud taken from above 3 cm shoot length and stem segments excised from below 3 cm shoot length were 5.0 mg /L and $1.0{\sim}3.0\;mg$ /L kinetin, respectively. Interesting phenomenon was observed, the direct formation of bulblets from the axilliary bud of cultured explants. Bulblet forming capacity in stem tissue was depended on stem age, young stem had high regeneration ability compared to old stem taken from above 10 cm shoot length. 1.0 mg /L kinetin was optimum concentration for the formation of bulblets from old stem segments. Stem tissue taken from underground growing plant was promoted coampare to shoot tips or bulb scale segments. Optimum concentration of sucrose was $5{\sim}7%$. Summariged above results revealed that effective explant for micropropagation was stem and /or node-bud tissue excised from less than 3 cm plant height compared to those of bulb scale segments which showed high contamination after culture. Maximum multiplication rate of young stem and /or node-bud segment was about 20 times. Kinetin requirement for stimulation of bulblet formation from cultured explant depended on source of explants but favorable levels of kinetin for organogenesis ranged from 1.0 mg /L to 5.0 mg /L.

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Morphogenetic Responses according to Media and Growth Regulators in vitro Culture of Eight Populus spp. (배지(培地) 및 생장(生長)호르몬에 따른 기내배양(器內培養)된 포푸라속(屬) 8개(個) 수종(樹種)의 기관형성반응(器官形成反應))

  • Kim, Chi Moon;Kwon, Ki Won;Moon, Heung Kyu;Lee, Jae Soon
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.205-212
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    • 1987
  • To examine the morphogenetic response, stem segments of 8 Populus spp. and 3 different explants of P. nigra var. italica were cultured on MS (Murashige and Skoog 1962) and WPM (Woody Plant Medium) medium containing various phytohormones. The results obtained were as follows: 1. Shoot regeneration and development from stem segment of 8 Populus spp. showed a quite difference according to the section and the species. All of the species of Leuce and Tacamahaca section did not form adventitious buds, while most of explants showed axillary or dormant bud elongation after 4 weeks. But P. nigra var. italica of Aigeiros section showed a successful adventitious bud formation (mean 5.4 buds per explant). 2. Leaf, petiole, and internode segment of P. nigra var. italica showed a quite differences according to media and ex plants upon the morphogenetic response. Adventitious bud formation from leaf was more abundant and readily initiated on the abaxial side than on the adaxial side. Mean number of 103 adventitious buds per explant was obtained from abaxial side of leaf segment cultured on WPM medium containing $0.2mg/{\ell}$ BAP for 5 weeks. 3. 2,4-D (2,4-dichlorophenoxy acetic acid) supplemented to media appeared to be negative upon the adventitious bud formation of P. nigra var. italica, while it promoted callus formation from all explants. Especially, NAA (${\alpha}$-naphtalene acetic acid) or NAA combination with BAP (6-benzylaminopurine) promoted root regeneration from the all explant of P. nigra var. italica in this study.

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Effective In Vitro Propagation from Pedicel Culture of Hippeastrum hybridum Hort. 'Dazzler' (아마릴리스 (Hippeastrum hybridum Hort. 'Dazzler') 소화경 배양에 의한 효율적 기내번식)

  • Kim Myung Jun;Kim Young Sook;Kim Hyun Soon;Ko Jeong Ae
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.382-389
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    • 2005
  • This study was conducted to establish the system of effective in vitro propagation by various explant sources culture of Bippeastrum hybridum Hort, 'Dazzler'. We tested the effects of optimal explant source, plant growth regulators on bulblet formation and plant regeneration. Callus was readily produced on the different tissues excised from floral buds whereas, bulbs and shoots were formed only on pedicel explants as compared with anthers, styles and ovaries. Pedicel is the best optimal explant for in vitro propagation. Two distinct pathways, organogenesis through callus and direct bulblet formation, could be recognized in pedicel culture. Up to the $80-100\%$ of bulblet formation and shoot organogenesis from the pedicel in fifteen days before anthesis were effectively induced by MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA. Plantlet regeneration was successfully achieved from pedicel-derived callus, via shoot bud induction or direct bulblet formation. The bulblets with blooming flower were produced within 2 years.