• Title/Summary/Keyword: in vitro plantlet regeneration

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In Vitro Plantlet Regeneration from Axillary Buds of Tilia amurensis Mature Trees and Clonal Variation in Tissue Culturability (피나무 성숙목(成熟木)의 액아배양(腋芽培養)에 의한 유식물체(幼植物體) 재생(再生)과 조직배양능력(組織培養能力)에 있어서의 클론간(間) 변이(變異))

  • Youn, Yang;Ohba, Kihachiro
    • Journal of Korean Society of Forest Science
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    • v.79 no.2
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    • pp.109-114
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    • 1990
  • The axillary buds of 15-year-old Tilia amurensis were cultured on Saito and Ide (IS), Murashige and Skoog (MS) media and woody plant medium (WPM) to establish an effective micropropagation method. Five levels of 6-benzylaminopurine (BAP) were tested. On IS medium and WPM addition of 1.0/l BAP enhanced shoot development and shoot elongation, whereas addition of 0.5/l BAP was effective on MS medium. A better results were obtained from WPM with 1.0/l BAP and MS with 0.1/l BAP. Developed shoots were subcultured on each basal media but with 0.2/l BAP, Multiple shoots were almost doubled in a month. Root formation could be enhanced at higher concentration of indole-3-butyric acid (IBA). Better rooting rate (83.3%) was achieved on a half-strength MS medium with 3.0 /l IBA. Regenerated plantlets were successfully transferred to soil. To investigate the clonal variation in shoot development and shoot elongation by axillary bud culturing, seven plus tree clones were tested, Clonal variation in tissue culturability among plus trees was recognized by the Duncan's multiple range test at the 5% level. Kang Won No. 12 showed the best response on WPM with 1.0/l BAP.

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Effect of Medium Composition on in vitro Propagation and Plantlet Regeneration from Nodal Explants of Cassava (Manihot esculenta Crantz)

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.16-16
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    • 2019
  • The Cassava (Manihot esculenta Crantz) is a perennial woody shrub cultivated mainly in the tropics for its starchy tuberous roots. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. Thus, a study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with single node after sterilization, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, single-node stem segments, approximately 1 cm in length, were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate was 55.6%, the shoot number and its length were 1.0/explant and 2.3 cm in the most favorable medium composition. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

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Factors Affecting the Production of In Vitro Plants from the Nodal Pieces of Chinese Yam (Dioscorea Opposita Thunb)

  • Shin, Jong-Hee;Kim, Sang-Kuk;Kwon, Jung-Bae;Lee, Bong-Ho;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.97-102
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    • 2004
  • This study was carried out to establish The regeneration of healthy seedlings from the nodal segment culture of Chinese yam (Dioscorea opposita cv. Danma), cultivated in Korea. Different explants such as leaves, petioles, roots and nodal pieces, excised from the in vitro grown seedlings of Chinese yam, were cultured on MS medium supplemented with various combinations of growth regulators. All the growth regulators used induced plantlet regeneration from the nodal segments at a high frequency, while there was no induction of shoot or callus from leaf, petiole or root tissues. The medium supplemented with 0.01mg/L NAA, 0.5mg/L BA, 0.5-1.0mg/L kinetin and without plant growth regulator was effective for shoot development of buds from the nodal segment culture. The concentration of BA and NAA was an important factor in the bud induction of buds from the nodal segments of Chinese yam. Nodal segments cultured on the medium containing 1.0mg/L NAA and 0.5-1.0mg/L BA gave the best response to bud formation. The addition of GA$_3$ to the culture medium suppressed shoot induction and growth, while it increased microtuber formation. The shoot growth and microtuber formation were also affected by medium strength and solidity. The MS basal medium containing 1 g/L gelrite was suitable for microtuber formation from the nodal segment of Chinese yam.

In vitro propagation of endangered species, Hylotelephium ussuriense (Kom.) H. Ohba (멸종위기종 둥근잎꿩의비름 (Hylotelephium ussuriense (Kom.) H. Ohba)의 기 내 증식)

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Kim, Ji-Ah;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.38-43
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    • 2014
  • To establish the system of in vitro plant regeneration, the different explants (stem with axillary bud and stem without axillary bud) of Hylotelephium ussuriense were cultured on the Murashige and Skoog's medium containing 6-benzylaminopurine (BA) and indolebutyric acid (IBA). The adventitious shoot induction was more effective in the stem with axillary bud explants than the stem without axillary bud explants, and was the best on MS medium containing 3.0 mg/L BA and 0.01 mg/L IBA. Frequency of plantlet growth was not significantly treated on MS and sucrose. Total chlorophyll contents under ventilation treatment were higher than those in control (non-ventilation). This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Effect of Growth Regulators of Plant Regeneration from Rhodiola sachalinesis leaf segments (홍경천 (Rhodiola sachalinesis)의 엽육 절편으로부터 식물체 분화에 미치는 생장조절제의 영향)

  • Bae Ki-Hwa;Yoo Ji-Ae;Yoon Eui-Soo
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.410-416
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    • 2005
  • Rhodiola sachalinensis has been used as a traditional medicine in Asia. We were germination in vitro seedling of grow naturally in Chang bai Moutain. And callus induction from leaf segments, treatmented plant regeneration in plant growth regulators (Auxins and cytokinins). We investigated optimal conditions for efficient plant regeneration through callus induction and shoots formation on medium with various kinds of growth regulators. Callus induction and adventitious shoots formation was achieved when cytokinin and auxin combinated to this experiment. Especially, there was the highest callus induction rates when we were used to 1 mg/L kinetin and 2 mg/L NAA $(98\%)$, Adventitious shoots formation wear obtained difference rate when cytokinin alone 1 mg/L BA $(96.6\%)$. And regenerated plantlet was acclimatized and transplanted to the soil, showed $100\%$ survival.

In vitro regeneration and the change of anatomical appearance in Poncirus trifoliata RAFIN. (탱자(Poncirus trifoliata RAFIN.)의 기내 재분화 및 조직학적 특성)

  • 박민희;이현화;장현규;이숙영;김홍섭
    • Korean Journal of Plant Resources
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    • v.12 no.2
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    • pp.107-119
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    • 1999
  • In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

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Cultural conditions affect somatic embryogenesis in Catharanthus roseus L. (G.) Don

  • Aslam, Junaid;Mujib, A.;Fatima, Samar;Sharma, M.P.
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.179-189
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    • 2008
  • We established an efficient plant regeneration system for Catharanthus roseus L. (G.) Don through somatic embryogenesis. Embryogenic callus was induced from hypocotyl of seed germinated in vitro. Somatic embryogenesis in Catharanthus has been categorized into three distinct stages: (1) initiation and proliferation of embryo; (2) maturation, and; (3) germination or plantlet conversion. Beside plant growth regulators, various stages of embryogenesis were screened for their response to a wide variety of factors (pH, gelrite, light, sugar alcohols, polyethyleneglycol and amino acids), which affect embryogenesis. All of the tested factors had a small to marked influence on embryogeny and eventual conversion to plantlets. The plantlets were acclimatized successfully in a greenhouse. To our knowledge, this is the first report describing a detailed study of various cultural factors which regulate embryogenesis in C. roseus. The results discussed in this paper may be used in mass propagation to produce medicinal raw material, and the embryo precursor cells could be used in genetic modification programmes that aim to improve the alkaloid yield as well.

Ultrastructural Characteristics of Developmental Stages During in vitro Regeneration in Citrus junos SIEB. (유자 (Citrus junos SIEB.) 의 발생단계에 따른 미세구조적 특성)

  • 박민희
    • Korean Journal of Plant Resources
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    • v.8 no.3
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    • pp.237-246
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    • 1995
  • In this study, the callus was induced and regenerated from the immature embryo and ultrastructural characteristics of developmental stages in Citrus junos SIEB, were investigated. The yellowish callus was induced by 5 to 6 week of culture of citrus. In proliferation callus after 6 weeks of culture, large vacuole was formed by fusion between adjacent small ones. In the non-embryogenic callus cultured for 12weeks, re-differentiated cells of callus showed the large nucleus with globular nucleus and amyloplast with large size of starches. In the embryogenic callus cltured for 14-16 weeks, the active exocytosis occurred in cells, secretory vesicles appeared on cell membrane and small particles from cytoplasm were released to intercelluar space. In the embryogenic callus cultured for 24 weeks, a sperical type of chloroplast bounded on cytoplasm by double membrane and typical grana was dispersed equally among matrix. In the normal plantlet after 26 weeks of culture, a lot of vessels and companion cells apperaed in the leaf cell of plantlet. In the normal plantlet after 30 weeks of culture, the immature leaf showed many small companion cells, sieve tubes and central vacuole. Also, the secondary vacuole protruded into the central vacuole and elongated chloroplasts near plasma membrane. In the matured plant habituated on the soil, palisada tissue composed of orderly arranged cells contained the nucleus in the center of the cell and large vacuoles on either side of the nucleus.

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Callus induction and high-efficiency plant regeneration via somatic embryogenesis in Papaver nudicaule L., an ornamental medicinal plant

  • Yang, Jing Li;Zhao, Bo;Seong, Eun-Soo;Kim, Myong-Jo;Kang, Won-Hee;Kim, Na-Young;Yu, Chang-Yeon;Li, Cheng Hao
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.261-267
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    • 2010
  • We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.

Correlation between In vitro Plant Regeneration and Polarity with Boxthorn (Lycium chinense Mill.) Seedlings (구기자나무 유식물체를 이용한 식물체의 재분화와 극성과의 관계)

  • Kwon Hye-Kyoung;Cho Yi-Yun;Yoon Eui-Soo
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.308-314
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    • 2006
  • This experiment was conducted out to investigate the effect of plant growth regulators on callus and shoot formation. The callus formation was effective on 1/2 MS medium containing 2,4-D, while shoot formation was suppressed. Shoot formation and differentiation were the highest in combination 0.1 mg/L of IAA and 0.1 mg/L of BA. The polarity of explants was investigated from cotyledon, which excised 20% of each basal and terminal parts. Formation of shoot was induced from excised ends of the basal part. In excised ends of the basal part, callus was induced vigorously and shoots were produced lately. Root induction was easily achieved in 1/3 MS medium from the adventitious shoot and more than 90% of regenerated plantlets acclimatized successfully and flowered normally.