• Title/Summary/Keyword: regenerated plantlet

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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.

Regeneration of symmetric protoplast fusion between cabbage (Brassica oleracea L.) and radish (Raphanus sativus L.) (양배추와 무의 동형 원형질체 융합을 이용한 식물체의 재분화)

  • In, Dong-Su;Song, Min-Jung;Jang, In-Chang;Min, Byung-Whan;Nahm, Seok-Hyeon;Shin, Jong-Sub;Lee, See-Woo;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.121-126
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    • 2008
  • Protoplasts from cabbage and radish were isolated and fused symmetrically by PEG treatment. The PEG treated mixture of high concentrated protoplasts produced lots of micro-calli after $2{\sim}3$ weeks. The microcalli developed to normal calli and shoots were regenerated from the calli. A total of 218 shoots were regenerated, but none of them contained the NWB-CMS specific DNA marker, indicating that the transfer of the radish NWB-CMS character into cabbage did not occur. However, ISSR analysis revealed that the cell fusion between protoplasts from radish and cabbage was occurred (3 out of 208 plantlet). The fused regenerants possessed the characteristics of source plants used for protoplast fusion. After vernalization, three regenerants were flowered with white petal color as seen in radish. Only three seeds were able to obtain from one regenerant by backcrossing with the cabbage pollen.

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.

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.

Plant Regeneration via Secondary Somatic Embryogenesis and Acclimatization in Panax ginseng (장뇌삼의 2차 체세포배 발생을 통한 식물체 유도 및 순화)

  • Lee, Su-Gwang;Kim, Ji-Hee;Kang, Ho-Duck
    • Journal of Korean Society of Forest Science
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    • v.97 no.1
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    • pp.127-133
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    • 2008
  • This study was conducted to establish the optimal condition for plant regeneration and acclimatization from somatic embryos of Panax ginseng. Cotyledon segments of Panax ginseng produced primary and secondary somatic embryos when cultured on MS and WPM media supplemented with 7% sucrose. To induce plantlet conversion, cotyledonary somatic embryos were cultured on WPM solid medium with $GA_3$ at various concentrations (1~30 mg/L) for 4 weeks. Plantlets were transferred to 1/2 WPM solid medium with $GA_3$ at various concentrations (0~5 mg/L) and 0.5% activated charcoal for shoot and root elongations. Elongated plantlets further developed into well-developed leaf and root system on 1/3 SH medium with 0.5% activated charcoal under ventilation condition for 5 months. The highest survival rate to soil was 75% when plantlets were regenerated on 1/3 SH medium without sucrose under ventilation condition.

Effect of BA and NAA on Adventitious Bud Induction From In Vitro Germinant Eucalyptus pellita (Eucalyptus pellita 기내발아묘의 부정아 유도에 미치는 BA 및 NAA 효과)

  • Kim Ji-Ah;Moon Heung-Kyu;Kang Ho-Duck
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.201-207
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    • 2005
  • This study was conducted to examine the effect of BA and NAA on adventitious bud induction from in vitro germinants E. pellita. The capacity of adventitious bud formation greatly depends on juvenility and explants origin; the more juvenile materials are the better ability to form adventitious buds even in in vitro raised plantlets. In case of in vitro germinants, 7 day old plantlets showed a better morphological response than did 14 day old ones in the induction of adventitious buds. The capacity to show morphological response was in decreasing order : cotyledons> petioles> roots. Ho adventitious buds formed when root segments were used as culture material. And optimum medium appeared to be MS + 0.5 mg/L BA and 0.2 mg/L NAA. Adventitious buds could be developed into multiple shoots and regenerated normal plantlets on DKW medium plus 0.2 mg/L BA and 0.01 mg/L NAA.

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.

Genetic polymorphism analysis of somatic embryo-derived plantlets of Cymbopogon flexuosus through RAPD assay

  • Bhattacharya, S.;Dey, T.;Bandopadhyay, T.K.;Ghosh, P.D.
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.245-252
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    • 2008
  • The genetic status of somatic embryo-derived plantlets of Cymbopogon flexuosus was examined by randomly amplified polymorphic DNA (RAPD) analysis. Auxins such as 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1-4 mg/l) were used in Murashige and Skoog (MS) medium for induction of calli from rhizomatous explants of Cymbopogon flexuosus. Optimum calli were induced on MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (2, 4-D) (3.5 mg/l) alone or in combination with $N^6-benzyladenine$ (2 mg/l). Somatic embryogenesis was achieved from long term calli when cultured on MS medium containing 2, 4-dichlorophenoxyacetic acid (2, 4-D) (2 mg/l) along with $N^6-benzyladenine$ (BA) (1-2 mg/l). Regeneration was achieved when freshly induced embryogenic calli were sub-cultured on MS medium supplemented with $N^6-benzyladenine$ (3 mg/l) alone. Long-term cultured embryos showed profuse minute rooting on regeneration medium supplemented with N6 -benzyladenine (3 mg/l). Microshoots were rooted in the presence of indole-butyric acid (IBA) (2 mg/l). DNA samples from the mother plant and 18 randomly selected regenerated plants from a single callus were subjected to RAPD analysis with 6 arbitrary decamer primers for the selection of putative somaclones. A total of 64 band positions were scored, out of which 19 RAPD bands were polymorphic. From genetic similarity coefficient based on RAPD band data sharing, it was found that the majority of the clones were almost identical or more than 92% similar to the mother plant, except CL2 and CL9 (66%) which showed highest degree of genetic change with CL2 and CL9 showing presence of two non-parental bands each.

Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by ${\gamma}$-irradiation ($^{60}Co$) of adventitious roots

  • Zhang, Jun-Ying;Sun, Hyeon-Jin;Song, In-Ja;Bae, Tae-Woong;Kang, Hong-Gyu;Ko, Suk-Min;Kwon, Yong-Ik;Kim, Il-Woung;Lee, Jaechun;Park, Shin-Young;Lim, Pyung-Ok;Kim, Yong Hwan;Lee, Hyo-Yeon
    • Journal of Ginseng Research
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    • v.38 no.3
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    • pp.220-225
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    • 2014
  • An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

Effect of Carbon and Nitrogen Source on Somatic Embryogenesis in Suspension Culture of Ligusticum chuanxiang Hort. (천궁의 현탁배양에서 탄소원과 질소원이 체세포배 형성에 미치는 영향)

  • Chae, Young-Am;Park, Sang-Un
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.1
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    • pp.44-50
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    • 1994
  • This study was carried out to select the appropriate medium(especially, carbon and nitrogen source ) for somatic embryogenesis in order to develop the rapid mass production system in suspension culture of chuanxiang Hort. Suitable medium for somatic embryo formation was MS medium. The half strength MS medium was effective for somatic embryo development. Sucrose was the most effective carbon source for somatic embryo formation, however, production of somatic embryos was reduced at higher concentration of sucrose. Effects of suger was the same as sucrose. Somatic embryo formation was higher as the decrease of $NH_{4}NO_3$, and optimum ratio of $KNO_3\;:\;NH_4NO_3$ was 825 : 238mg /1. Regenerated plant was obtained in MS basal medium and survival late of plantlet was 60-70% after transplanted directly to the vermiculite.

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