• Title/Summary/Keyword: Callus Induction

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Secondary Products in Cell Suspension Culture of Salix koreensis (버드나무(Salix koreensis) 현탁배양(懸濁培養) 세포(細胞)의 대사산물(代謝産物))

  • Park, Young Goo;Shin, Dong Ill;Lee, Sang Goo
    • Journal of Korean Society of Forest Science
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    • v.78 no.2
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    • pp.198-208
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    • 1989
  • Cell suspension cultures for Salix koreensis was well established at the supplements of 2, 4-D with cytokinin particulary the combination of 1.0 mg/l 2, 4-D with 0.1 mg/l of zeatin. These combined rates of phytohormones are also effective to callus induction from S, koreensis leaf and its multiplication. Cultured media exhibited the great inhibitory effect on the germination of rice, barnyard grass and lettuce seeds, indicating the presence of biologically active substances in media. Several phenolic compounds such as pyrogallol, sinapic acid, cinnamic acid, tannic + gallic and p-chlorobenzoic acid were detected in the cell suspension culture. The inhibitory effect exhibited by cultured media may be partly attributed to these phenalic compounds.

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In vitro shoot initiation of Artocarpus heterophyllus Lam. (Jak Fruit) Effect of the explant type and the season of explant collection

  • Kahk, Kasturiarachchi;Wtpsk, Senarath;Lee, Kui-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10a
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    • pp.9-18
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    • 2003
  • A method for rapid propagation of mature Jack fruit was developed. Four types of explants (mature embryos, apical meristems of young seedlings, apices from mature plants and nodal segments) were used. It has been found 88% of young apical meristems produced shoots in Campbell and Durzan (CD) medium compared to 60% in Murashige and Skoog (MS) medium. Only 1/3 of them produced multiple shoots. Shoot idtiation from nodal segments was very rare. Mature apices produced callus. Although removed of the sheathing cover around mature buds enhanced the shoot initiation but success rate was low in growth regulator free medium. Embryos respond to the CD medium but not to the MS medium. Embryos from seeds soaked in water for 24 hours produced shoots after 8 weeks of incubation and the success rate was 70% while embryos from dry seeds only produced roots. There was no significant effect of cold storage (refrigeration) for 7 days on shoot initiation from mature embryos (65%) but the ability for shoot induction declines with storage time (55% after 21 days of cold storage). Mature axillary buds were established in Modified Campbell and Durzan (CD) medium supplemented with 0.5mg/1 and IBA. There was a significant difference in the growth performance of shoots according to the period of the year in which explants were collected. Highest (60%) was observed in November-January period. It was only 30% when the explants were collected in February-April or May-July and decreased to 20% in August-October. The shoots produced in November-January showed a higher vigor than those produced in other months. Since Jak fruit show seasonal changes in fruit bearing and shedding of leaves, it can be suggested that the difference in growth performances of tissues cultured in artificial culture media would have been affected by endogenous rhythms.

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Propagation by In Vitro Zygotic Embryos Cultures of the Quercus myrsinifolia

  • Choi, Eun ji;Yong, Seong Hyeon;Seol, Yu Won;Park, Dong Jin;Park, Kwan Been;Kim, Do Hyun;Jin, Eon Ju;Choi, Myung Suk
    • Journal of Forest and Environmental Science
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    • v.37 no.4
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    • pp.323-330
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    • 2021
  • Zygotic embryo culture was performed to propagate evergreen oak, Quercus myrsinifolia, which has recalcitrant seeds and is difficult to propagate by cuttings. Zygotic embryos appeared in WPM medium after 14 days, and after 56 days, they developed into complete plants with cotyledons and roots. The medium suitable for zygotic embryo culture was 1/4 WPM medium, showing a shoot growth of 2.43 cm and root growth of 8.7 cm after 8 weeks of culture. As a result of investigating the effect of GA3 on the growth of plants germinated from zygotic embryos through GA3 treatment, the best growth was shown in 0.5 mg/l GA3 treatment. The in vitro rooting and growth of IBA-treated zygotic embryo-derived plants were good in the 0.5 mg/l IBA treatment and rooting and shoot growth were not observed at higher concentrations. And the callus induction rate also increased as the concentration of IBA increased. Plants grown in vitro were transferred to a plastic pot containing artificial soil and acclimatized in a greenhouse for about 4 weeks, resulting in more than 90% survival. As a result of this study, the zygotic embryo culture method was confirmed to be effective for mass propagation of Q. myrsinifolia. The results of this study are expected to contribute significantly to the mass propagation of elite Q. myrsinifolia.

Tissue-cultured regeneration and ecological values in major bamboo species

  • Sharma, Avinash;Manpoong, Chowlani;Gohain, Anwesha;Pandey, Himanshu;Padu, Gompi;Aku, Hage
    • Journal of Ecology and Environment
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    • v.46 no.3
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    • pp.218-242
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    • 2022
  • Background: Promising specific growth regulators are employed in the tissue cultures of various bamboo species. Specific natural hardening mixtures support the acclimatization and adaptation of bamboo under protected cultivation. Results: The growth regulators like 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Naphthaleneacetic Acid (NAA), Thidiazuron (TDZ), 6-Benzylaminopurine (BAP), Kinetin, Gelrite, Benzyl Adenine (BA), Indole Butyric Acid (IBA), Coumarin, Putrescine, Gibberellic acid (GA3), Indole Acetic Acid (IAA) has been widely used for callus induction, root regeneration and imposing plant regeneration in various species of bamboo such as Bambusa spp. and Dendrocalamus spp. Different combinations of growth regulators and phytohormones have been used for regenerating some of the major bamboo species. Natural hardening materials such as cocopeat, vermicompost, perlite, cow dung, farmyard manure, compost, soil, garden soil, and humus soil have been recommended for the acclimatization and adaptation of bamboo species. Standard combinations of growth regulators and hardening mixtures have imposed tissue culture, acclimatization, and adaptation in major bamboo species. Conclusions: Bamboo contributes to soil fertility improvement and stabilization of the environment. Bamboo species are also involved in managing the biogeochemical cycle and have immense potential for carbon sequestration and human use. This paper aims to review the various growth regulators, natural mixtures, and defined media involved in regenerating major bamboo species through in vitro propagation. In addition, the ecological benefits of safeguarding the environment are also briefly discussed.

Production of ${\beta}-sitosterol$ by Cell Suspension Culture of Chrysanthemum coronarium L. (쑥갓세포의 현탁배양에 의한 ${\beta}-sitosterol$ 생산)

  • Kim, Hyun-Chul;Chung, Ha-Young;Lee, So-Youn;Chung, Ho-Yong;Kim, You-Jung;Baek, Nam-In;Kim, Soung-Hoon;Choi, Geun-Won;Kim, Dae-Keun;Kwon, Byoung-Mok;Park, Mi-Hyun;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.425-430
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    • 2005
  • [${\beta}-sitosterol$] is a plant sterol that reduces cholesterol levels and inhibits the growth of human prostate and colon cancer cells. Optimal conditions for ${\beta}-sitosterol$ production were examined from cell suspension cultures of Chrysanthemum coronarium L. The callus induction was optimal in MS medium containing 1 mg/l NAA and 1 mg/l BAP. Cell suspension culture was also established from the callus. Optimal ${\beta}-sitosterol$ production was obtained when the cells were cultured at an initial density of 2 mg DCW/l in MS medium containing 1 X sucrose (30 mg/l), 1 X nitrogen (1900 mg/l $KNO_3$, 1650 mg/l $NH_4NO_3$), and 1 X phosphate source (170 mg/l). In cell suspension cultures of C. coronarium L. using shake flasks, the peak content of ${\beta}-sitosterol$ was $150{\mu}g/g$ DCW. In cell suspension cultures of C. coronarium L. using an air-lift bioreactor, the maximum ${\beta}-sitosterol$ content of $143.8{\mu}g/g$ DCW was obtained at an air-flow rate of 100 cc/min.

The Establishment of Nitrogen Fixation by Cultured Cell-Rhizobium Association Through Tissue Culture Technique in Soybean (대두조직배양세포(大豆組織培養細胞) - Rhizobium에 의(依)한 질소고정력(窒素固定力))

  • Kang, Sang Jae;Park, Woo Churl
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.27-35
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    • 1986
  • This experiment was carried out to elucidate the factor of nitrogenase formation and to establish the nitrogen fixation system in mixed culture of cultured cells and rhizobia through tissue culture technique using three soybean varieties, Hwangkeum, Namcheon and D 68-0099 as host plants. The results obtained were as follows; The callus was induced in embryo and radicle, but not in hypocotyl. The most favorable callus induction was caused by the individual application of 2,4-D and NAA at the concentration of 2mg/1 and 4mg/1, respectively, but in case of treating both 2,4-D and kinetin, that was done at the concentration of 0.2mg(2,4-D)/0.05mg(kinetin)per liter. The growth of cultured cell was good at the concentration of 2.0mg(2,4-D)/1 and 0.2mg(2,4-D)/0.05mg(kinetin)per liter. When cultured cells were inoculated with R. japonicum 019 and 011, their growthes were considerably inhibited. The addition of single amino acid inhibited the growth of cultured cells. Hwangkeum was inhibited considerably by methionine and leucine. The inhibition of growth by single amino acid can be abolished by the addition of certain amino acids. The differentiation of adventitious root was good at the concentration of 2.0mg 2,4-D and 0.2mg 2,4-D/0.05mg kinetin per liter. Of three host plants tested with 25 R. japonicum strains, Hwangkeum had affinity for 10 strains, Namcheon for 7 strains and D68-0099 for none. The nitrogen fixing abilities of Hwangkeum and Namcheon caused by cultured cell-Rhizobium association were high in strain 019, 007, and in 007 mixed with 119, respectively.

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Multi-secondary Somatic Embryogenesis and Plant Regeneration from Hypocotyl Cultures of Alfalfa (Medicago sativa L.) (알팔파의 하배축으로부터 다량의 이차 체세포배 발생과 식물체 재분화)

  • Won, S.H.;Lee, B.H.;Kim, K.Y.;Lee, H.S.;Lee, H.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.273-280
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    • 1999
  • Hypocotyl explants of Medicago saliva cv. Vernal were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of growth regulators. After six weeks of culture, somatic embryos were formed from calli on MS medium containing $4mg/{\ell}$ 2,4-D and $0.1mg/{\ell}$ kinetin, or $4mg/{\ell}$ 2,4-D and $0.5mg/{\ell}$ kinetin. The mature somatic embryos were developed to plantlets when subcultured on MS basal medium. In order to obtain secondary somatic embryogenic calli, cotyledon of regenerated plantlets were cultured on a callus induction medium. Embryogenic calli were formed on MS medium containing $4mg/{\ell}$ 2,4-D alone. For induction and development of secondary somatic embryogenesis, the embryogenic calli were transferred to MS basal medium containing either 2,4-D or NAA. Multi-secondary somatic embryogenesis was the most effective on MS basal medium with $0.1mg/{\ell}$ 2,4-D. The rate of secondary somatic embryo formation of regenerated plants was 18 times higher than that of seed grown plants. The mature secondary somatic embryo were germinated into plantlets on MS basal medium after six weeks of culture.

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Effects of Plant Growth Regulators on In Vitro Germination and Organ Formation of Wild Angelica gigas N. (야생 참당귀(Angelica gigas N.)의 기내발아 및 기관유도에 미치는 PGRs의 영향)

  • Lee, Su-Gwang;Cho, Won-Woo;Lee, Song-Hee;Park, Kwang-Woo;Choi, Kyung;Kang, Ho-Duck
    • Korean Journal of Plant Resources
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    • v.25 no.4
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    • pp.456-464
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    • 2012
  • This study was conducted to establish the in vitro optimal condition for seed germination and organogenesis of wild Angelica gigas. The experiment was evaluated the effects of $GA_3$ for pre-treatment with different periods of time (0h, 24h, 48h, 72h) and followed the treatment of seeds by control, scarification and methanol-heating method. As a result, the highest rates (15%) of seed germination was shown under the treatment without soaking of $GA_3$ and methanol-heating treatment. The seed germination was highly increased 60% under the condition of treatment on ultrasonic waves (frequency 80 KHz) with methanol-heating treatment including 0.1 mg/L $GA_3$. The highest callus induction rate was obtained from in vitro germinated stem, root and hypocotyl on the MS medium with 1.0 mg/L NAA and 0.5 mg/L BA. The highest percentages of shooting (50%) and rooting (85%) induction were observed in hypocotyl and root cultured on PGRs free medium and 0.1 mg/L NAA, respectively. In addition, somatic embryogenesis was observed from stem (1.0 mg/L 2,4-D) and hypocotyl (0.1 mg/L NAA).

Plant Regeneration of Major Cultivars of Sweetpotato (Ipomoea batatas) in Korea via Somatic Embryogenesis (체세포배발생을 통한 국내 주요 고구마 품종의 식물체 재분화)

  • Kwon, Eun-Jeong;Kwon, Suk-Yoon;Kim, Moon-Za;Lee, Joon-Seol;Ahn, Young-Sup;Jeong, Byeong-Choon;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.189-192
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    • 2002
  • An efficient plant regeneration system of major cultivars of sweetpotato (Ipomoea batatas (L.) Lam.) in Korea via somatic embryogenesis was established. Embryogenic calli were formed from shoot apical meristems of sweetpotato cultivars when cultured on LS medium supplemented with 1 mg/L auxin (2,4-D, picloram, dicamba). Among three kinds of auxin, 1 mg/L 2,4-D showed the highest embryogenic calli induction rate. After 4 weeks of cultures on LS medium supplemented with 1 mg/L 2,4-D, embryogenic calli induction rates of Sinhwangmi, Zami, Yulmi, and White Star were 86%, 78%, 76%, and 80%, respectively. Upon transfer onto LS basal medium, most of somatic embryos developed into plantlets. Regenerated plantlets were transplanted to potting soil and grown to mature plants in a greenhouse.

Acquirement of transgenic rose plants from embryogenic calluses via Agrobacterium tumefaciens (배발생 캘러스를 이용한 아그로박테리움 매개형질전환 장미 식물체 획득)

  • Lee, Su-Young;Lee, Jung-Lim;Kim, Won-Hee;Kim, Seung-Tae;Lee, Eun-Kyung
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.511-516
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    • 2010
  • The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.