• Title/Summary/Keyword: Callus Induction

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Plant Regeneration from Cambium Callus of Ailanthus altissima Swingle (가중나무의 형성층(形成層) Callus에서 식물체(植物體) 재분화(再分化))

  • Lee, Sang Goo;Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.78 no.4
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    • pp.412-418
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    • 1989
  • The stem segments of Ailanthus altissima were cultured on the Murashige & Skoog's medium(1962) supplemented with 0.1 mg/l BAP and 1.0 mg/l 2, 4-D for callus induction and proliferation, Shoot primordia were observed as greenish regions on the surface of yellow-brown calli about 8 weeks after culture. Shoot primordia were selected and transferred to the MS media containing various combination of BAP and 2, 4-D. Among these combinations the shoot primordia cell clusters on the medium added to 0.5mg/l BAP and 0.01mg/l 2, 4-D exhibited the highest number of shoot formation. These shoots were successfully transferred on the solid MS medium with no growth regulators for the rootings.

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Effects of $GeO_2$ and Citric Acid on Germanium Content of Callus and Plant in Angelica koreana MAX (강활(羌活)의 캘러스 및 식물체(植物體) 중(中) Ge함량(含量)에 미치는 $GeO_2$와 Citric Acid의 영향(影響))

  • Park, Byoung-Woo;Lee, Joong-Ho;Kwon, Tae-Oh
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.2
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    • pp.101-108
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    • 1996
  • This study was conducted to investigate the effect of growth regulators and $GeO_2$ on the induction and proliferation of callus and the effect of $GeO_2$ and citric acid on the Ge content of callus from explants and plant, Angelica koreana Max. The results obtained were summarized as follows. The callus induction was most effective on MS (Murashige and Skoog) medium containing 1. 0ppm 2, 4 - D with petiolule. The proliferation of callus was most effective at 2. 0ppm 2, 4 - D on the medium, at 2. 5ppm $GeO_2$ on the medium containing 2. 0ppm 2, 4 - D, and at $0.\;1{\sim}1mM$citric acid on the medium at pH6 containing 2. 0ppm 2, 4 - D and 2. 5ppm $GeO_2.$ The more $GeO_2$ in MS medium up to 20ppm, the more Ge content in callus. Ge content in callus was highest when the medium was supplemented with 0. 1mM citric acid and the pH of medium was low. The Ge content in plant was high in order of leaf > root > stem. Application $GeO_2$ to the soil increased Ge content in plant and application of 1mM citric acid with $GeO_2$ resulted in increasing Ge content highest in plant, but application more than l0mM citric acid resulted in Ge content decreased. Application of $GeO_2$ increased Ge content in callus and plant but had a tendency to decrease some mineral content, on the other hand application of $0.\;1{\sim}1mM$ citric acid with $GeO_2$ had a tendency to increase mineral content.

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Plant regeneration via callus culture of sea-milkwort (Glaux maritima L.) (갯봄맞이(Glaux maritima L.)의 캘루스 배양을 통한 식물체 재분화)

  • Han, Jeung-Sul
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.171-177
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    • 2017
  • A callus-mediated regeneration protocol for sea-milkwort, an endangered coastal plant species in South Korea, is reported here. The explants of in vitro-plantlets generated from a node culture revealed distinguishable responses in callus induction depending on genotype, explant source, light condition, and 2,4-D concentration. Especially, continuous darkness exclusively facilitated callus induction from explants prior to other treatments. The calli initiated on the media with 2,4-D ranging from 0.1 mg/L to 3.0 mg/L in the dark vigorously proliferated when subcultured on the same media in continuous darkness. Given 1.0 mg/L zeatin in addition to darkness to the calli of the 'Pistachio' genotype, normal adventitious shoots were only regenerated from nodular structures that formed earlier from the calli at the frequency of 24.4 percent. Regenerated shoots easily grew into plantlets with roots and green color on a phytohormone-free MS medium under lighted condition, that were used for node culture as plant materials. Node culture effectively multiplied plantlets in accordance with protocol by Bae et al. (2016). Acclimatized plantlet clusters developed mature plant clusters under inland environment, followed by flowering the following April. Results were merged with node culture protocol suggested by Bae et al. (2016), which, as an in vitro propagation system for sea-milkwort, may contribute to natural habitat restoration.

Effect of Plant Growth Regulators and Medium Supplements on Plant Regeneration of Kentucky Bluegrass (식물생장조절물질과 배지첨가물질이 켄터키 블루그래스의 식물체 재분화에 미치는 영향)

  • Lee Sang-Hoon;Lee Ki-Won;Kim Do-Hyun;Lee Dong-Gi;Won Sung-Hye;Kim Ki-Yong;Lee Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.26 no.2
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    • pp.69-76
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    • 2006
  • To optimize tissue culture responses for genetic transformation of Kentucky bluegrass, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar 'Newport' as explant tissues. The optimal concentration of 2,4-D (2.4-dichloro phenoxy acetic acid) for the induction of embryogenic callus from mature seed was 3 mg/L. Plant regeneration frequency was 54% when embryogenic callus was cultured on the regeneration medium supplemented with 1 mg/L 2,4-D and 3 mg/L of BA (6-benzyladenine). Addition of 1 g/L of casein hydrolysate and 500 mg/L of L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 60.8% and 58.3%, respectively. Regenerated plants were grown normally when shoots transplanted to the soil. A rapid and efficient plant regeneration system established in this study. We suggest that the results may be useful for molecular breeding of Kentucky bluegrass through genetic transformation.

Response to Anther and Tissue Cultures of Corn, Pearl Millet and Buckwheat Genotypes (옥수수, 진주조, 메밀의 약 및 조직배양 반응)

  • Keun-Yong Park;Rae-Kyung Park;Byeong-Han Choi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.2
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    • pp.142-146
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    • 1989
  • Anther and/or tissue culture of cross pollinated crops would be very important because it can result in the direct use of haploids or doubled haploids for developing superior hybrids or varieties. The objective of the study was to investigate the response frequencies in anther and/or tissue-cultured hybrids of corn. pearl millet and buckwheat to identify agronomically acceptable germplasm of the crops. 27 crosses of corn inbred lines were evaluated by plating their anthers on N6. MS and Yu-Pei media. Two genotypes of FR1l41/FR16 hybrid cultured on N6 medium and Fla 2BT73/S6013 hybrid cultured on N6 medium responded with one anther producing calli when plated after 5$^{\circ}C$ low temperature treatment for one week. Immature embryos of corn hybrid Suwon 19 responded producing calli that were regenerated to plants at a 8.6 percent success rate. Of the 20 corn hybrids. immature tessels of FR1l41/FR16. B68/A1l6N//KS15. KS16/KS17. GA209/DB578 and SDB126/GA209 crosses responded at a relatively higher success rate producing calli that were regenerated to plants. In tissue culture of elongating culms of pearl millet x Napier grass interspecific hybrid. 2.5-4.0mm long pieces of the culm were good for callus induction resulting in higher success rate. The epicotyl of buckwheat was very good for tissue culture. and the node produced the plants regenerated directly without callus induction on the B5 medium containing I ppm BA and 0.05 ppm IBA. There were great differences in response to anther and/or tissue culture of corn, pearl millet and buckwheat due to genotype x medium and environment interactions.

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Plant regeneration through the callus culture induced from bulb scales of an endangered species Lilium cernum Komarvo. (멸종위기종인 솔나리(Lilium cernum Komarvo.)의 인편 유래 캘러스 유도 및 기내 식물체 재분화)

  • Bae, Kee-Hwa;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.65-71
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    • 2013
  • Lilium cernum Komarvo. is an important endangered plant belonging to the family Liliaceae. A method was developed for the rapid micropropagation of L. cernum through plant regeneration from bulb scales explant-derived calli. The bulb scales segments were cultured on Murashige and Skoog (MS) medium supplemented with 0, 0.5, 1.0, $3.0mg{\cdot}L^{-1}$ kinetin and 0, 0.1, 0.5, $1.0mg{\cdot}L^{-1}$ NAA or 2,4-D for callus induction. In media with $0.5{\sim}3.0mg{\cdot}L^{-1}$ kinetin and $0.1{\sim}1.0mg{\cdot}L^{-1}$ NAA and 2,4-D, 95~100% of explants produced callus. They were then transferred to MS medium supplemented with various concentrations of NAA (0, 0.01, 0.05 and $1.0mg{\cdot}L^{-1}$) in combination with BA (0, 1.0 and $2.0mg{\cdot}L^{-1}$) for bulbet formation. Bulbet induction (78%), weight (468 mg) and size (15.5 mm) were obtained the highest on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. In vitro frequency of plant regeneration was not significantly treated in strength of MS and sucrose concentration. Chlorophyll contents in 1/2MS with $50g{\cdot}L^{-1}$ sucrose treatments were higher than those in control and another treatment. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Effect of growth regulators on In Vitro mass propagation of Haworthia maughanii (생장조절제가 하월시아 만상(Haworthia maughanii)의 기내 대량증식에 미치는 영향)

  • Kim, Youn Hee;Kim, Hye Hyeong;Lee, Gee Young;Lee, Jae Hong;Jung, Jae Hong;Delgado-Sanchez, Pablo;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.369-374
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    • 2018
  • The purpose of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shoot and rooting for the in vitro mass production of Haworthia maughanii. To determine suitable parts of the plant for callus induction, the leaves, flower bloom and flower stalks were cultured in MS medium at different concentrations of $0{\sim}2mgL^{-1}$ NAA and $0{\sim}2mgL^{-1}$ TDZ, respectively. All of the parts showed 100% callus formation rate at $NAA\;1mgL^{-1}$ and $TDZ\;1mgL^{-1}$ treatment, $NAA\;2mgL^{-1}$ and $TDZ\;2mgL^{-1}$ treatment and NAA 1 to $2mgL^{-1}$, respectively. While the rate of callus formation was high in all parts, the leaves were the most efficient to obtain most culture parts. $NAA\;0.1mg\;L^{-1}$ and $BA\;0.1mg\;L^{-1}$ treatments were the most effective in shoot formation with 22.0 shoots. In addition, multiple shoot propagation showed 16.3 shoots, the highest, with $NAA\;0.1mg\;L^{-1}$ and $BA\;0.1mg\;L^{-1}$ treatments. These results led us to speculate that the optimization of culture conditions was responsible for the mass propagation for in vitro cultures of Haworthia maughanii.

Plant Regeneration from Cell Suspension Culture Using Leaf Callus in Actinidia deliciosa X A. arguta Clone 118 (양다래X다래 클론 118의 엽조직 캘러스를 이용한 세포 현탁배양으로부터 식물체 유도)

  • Kim Yong-Wook;Moon Heung-Kyu
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.287-292
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    • 2005
  • Calli were induced by culturing the leaf segment of Actinidia deliciosa ${\times}$ A. arguta clone 118 on MS medium supplemented with 0.5 mg/L 2,4-D, 0.1 mg/L NAA and 0.05 mg/L BA for 8 weeks in light condition. The induced calli were inoculated in liquid MS medium containing 0.5 mg/L 2,4-D, 0.1 mg/L NAA, 0.05 mg/L BA and 3% sucrose to establish cell suspension culture. The cells at the exponential stage and the stationary stage could be observed between 5-11 days and after that 12 days in culture, respectively. The fresh weight of callus induced from the suspended cells did not vary much among the media containing eight different combinations of plant growth regulators tested. The highest frequency of shoot induction (88.3%) was observed in MS medium containing 2.0 mg/L zeatin. Either BA or zeatin mixed with thidiazuron (TDZ) seemed to be effective in shoot induction. The induced shoots were transferred to MS medium containing 0.2 mg/L zeatin for further shoot growth. And then the shoots were transferred to Standardi (ST) medium containing 1.0 mg/L indolebutyric acid (IBA) for rooting. Plantlets could be obtained through cell suspension culture of Actinidia deliciosa ${\times}$ A. arguta clone 118.

Stable Transformation via Callus Formation and Rhizogenesis from the Cultures of Hypocotyl Explant of Chinese Cabbage (배추의 배축절편으로부터 캘러스와 뿌리 발생을 통한 안정적 형질전환)

  • Cho, Mi-Ae;Kim, Choon-Ae;Min, Sung-Ran;Ko, Suck-Min;Liu, Jang-Ryol;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.139-144
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    • 2007
  • Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

Multiple Shoot Induction from Radicle-derived Callus and in Vitro Propagation of Silene Acaulis Subsp. Arctics (극지식물 Silene acaulis subsp. arctica의 유근 유래 캘러스로부터 다신초 유도와 기내 증식)

  • Seo, Hyo-Won;Yi, Jung-Yoon;Park, Young-Eun;Kang, Sung-Ho;Chung, Ho-Sung;Kim, Ji-Hee
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.303-307
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    • 2006
  • We describe here an efficient in vitro propagation method of Silene acaulis subsp. arctica (Caryophyllaceae), one of the higher arctic angiosperms, through the multiple shoot regeneration after callus induction from the radicle. The seeds of S. acaulis subsp. arctica collected from Svalbard, the Norwegian Arctic, were germinated and calli were induced from the radicle on solid MS media supplemented with 0.25mg/L 2,4-D and 1mg/L $GA_3$ at both $10{\pm}1^{\circ}C\;and\;23{\pm}1^{\circ}C$ Two weeks after callus induction, the multiple shoots were efficiently regenerated on the MS media supplemented with 0.25 g/L BA and 0.05mg/L HPh. The total biomass increment of regenerated shoots increased most efficiently of S. acaulis subsp. afctica was showed the maximum efficiency in at $23{\pm}1^{\circ}C$ on 1/2 MS salt strength. The multiple regenerated plantlets of S. acaulis subsp. arctics were grown to normal plants on soil.