• Title/Summary/Keyword: Embryogenic suspension culture

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In vitro Regeneration of Phragmites australis through Embryogenic Cultures

  • Lee Jeong-Sun;Kim Chang-Kyun;Kim In-Sung;Lee Eun-Ju;Choi Hong-Keun
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.21-25
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    • 2006
  • Phragmites australis (reed) has received much attention as being one of the principle emergent aquatic plants for treating industrial and civil wastewater. Plant regeneration via plant tissue culture in p. australis was investigated. Three types of callus were identified from seeds on N6 medium plus 4.5 UM 2,4-dichlorophenoxyacetic acid (2,4-D). Yellow compact type showed the best redifferentiation, whereas white compact type and yellow friable were not competent to differentiate into plane. Solid medium culture was better than liquid suspension culture for enhancing callus growth when N6 medium supplemented with 4.5 ${\mu}M$ 2,4-D was used. Phytagel, as a gelling agent, was superior to agar in plant regeneration on N6 medium, supplemented with 9.4 ${\mu}M$ kinetin and 0.54 ${\mu}M$ $\alpha$-naphthaleneacetic acid (NAA). Transfer of the plantlets regenerated from kinetin and NAA-supplemented N6 medium to growth regulator-free MS medium enhanced the further development of the plantlets. Plantlets on subsequently grown to maturity when tansferred to potting soil. The regenerated plants exhibited morphologically normal. The system for plant regeneration of P. australis enables to propagate elite lines on a large scale for water purification in the ecosystem

Mass Production of Eleutherococcus senticosus Plants through in vitro Cell Culture (세포 배양을 통한 가시오갈피(Eleutherococcus senticosus) 묘목의 대량 생산 시스템 개발)

  • Han, Jeong-Yeon;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.167-172
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    • 2003
  • Immature zygotic embryos of Eleutherococcus senticosus seeds matured rapidly within one month when the seeds comprising zygotic embryos were pieced to small size and cultured on 1/2 MS medium. Frequency of somatic embryos formation was declined rapidly when the zygotic embryos germinated and grew to plantlets. Embryogenic cells were induced by consecutive subculture of somatic embryos on MS medium with 1.0mg/L2,4-D. After heart-shaped somatic embryos were induced by suspension culture, these embryos were plated onto petri dish to support maturation of embryos. Germination of embryos occurred on medium with 5mg/L GA$_3$and transferred to culture bowl to stimulate the further growth. Frequency of soil survival of plantlets was influenced by soil mixture (perlite and peatmoss). The suitable combination of perlite and peatmoss was 1:5, and the soil survival rate was 78% after 4 months. The soil transferred plantlets were over-wintered in field condition after defoliation. New year sprouting of plants was achieved successfully and they grew to adult plants. These results indicate that the systematic procecure of plant production in E. senticosus for micro propagation.

High frequency plant regeneration from zygotic-embryo-derived embryogenic cell suspension cultures of watershield (Brasenia schreberi)

  • Oh, Myung Jin;Na, Hye Ryun;Choi, Hong-Keun;Liu, Jang Ryol;Kim, Suk Weon
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.87-92
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    • 2008
  • An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

Somatic Embryogenesis and Plant Regeneration in Suspension Cultures of Aralia elata S (두릅캘러스의 현탁배양에서 체세포배발생과 식물체 재분화)

  • 장한호;박철호;이윤수;신영범
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.167-171
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    • 1994
  • This study was carried out to investigate the possibility of plant regeneration through somatic embryogenesis in suspension culture of Aralia elata S. Callus was induced from the explants of leaf and petiole cultured in the MS media containing 2,4-D and TDZ. More embryogenic calli were formed from petiole and with combination treatment of 24-D and TDZ. The quarter strength MS medium was effective for increasing number of somatic embryos. Mannitol supplemented to the quarter strength MS medium, reduced somatic embryo formation but inositol increased. Normal plantlets(86%) were regenerated from mature somatic embryos in MS basal medium and 50% of those survied when transplanted to the vermiculite in greenhouse.

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Expression of Phosphinothricin Acetyltransferase Gene in Transgenic Rice Plants (형질전환체 벼에서 phosphinothricin acetyltransferase 유전자 발현)

  • Lee, Soo-In;Lee, Sung-Ho
    • Journal of Life Science
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    • v.14 no.2
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    • pp.368-373
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    • 2004
  • We have obtained fertile transgenic rice plants resistant to the broad spectrum herbicide Bast $a^{(R)}$ (active ingredient phosphinothricin, PPT) by PEG-mediated transformation procedure. The plasmid pCaMV35S::Bar was used to deliver the bar gene into embryogenic suspension culture-derived protoplasts of rice (Oryza sativa L.). Transformed plants were regenerated and selected on medium containing 15 mg/l of phosphinothricin. Stable integration and expression of the bar gene in transgenic rice plants was confirmed by Southern and Northern blot analysis. Transgenic $R_1$ plants were also confirmed by assays for phosphinothricin acetyltransferase (PAT) activity. The bar gene was expressed in the primary transgenic rice plants and in the next generation progeny, in which it showed a 3 : 1 Mendelian inheritance pattern. Transgenic $R_1$ and $R_2$ plants were resistant to the herbicide Bast $a^{(R)}$ when sprayed at rates used in field practice.ice.

Several Factors on Bulblets Regeneration from Callus Culture in Lilium longiflorum 'Celia' (백합 'Gelia' 캘러스로부터 자구 재분화에 미치는 제요인)

  • 박소영;김시동;신세균;이철희;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.183-188
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    • 1997
  • Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

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Regeneration of Fertile Transgenic Rice Plane from a Korean Cultivar, Nakdongbyeo (한국 재배종 낙동벼에서 임성 형질전환식물체의 재분화)

  • Soo In LEE;Hyun Jin CHUN;Chae Oh LIM;Jeong Dong BAHK;Moo Je CHO
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.175-182
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    • 1995
  • Rice is one of the most successful monocot in regenerating fertile and genetically stable transgenic plants. However there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper we first demonstrate that a Korean variety Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphotransferase) and GUS ($\beta$-glucuronidase) genes in separate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiment, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hy $g^{R}$ calli. The average efficiency of plantlet regeneration was apprbximately 27%. Based on the GUS activities of hygromycin resistant calli, ca.35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and Rl seeds were obtained. By examining the in siぉ activity of GUS in Rl seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the Rl generation.n.

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Plant Production from Desiccated Somatic Embryos of Acanthopanax chiisanensis (지리오가피 (Acanthopanax chiisanensis) 체세포배의 건조처리를 통한 식물체 증식)

  • Lee, Kang-Seop;Bang, Keuk-Soo;Choi, Yong-Eui;Ahn, Byung-Yong
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.381-385
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    • 2003
  • An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100%) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA+0.5mg/L BA or 0.5 mg/L NAA+0.5mg/L BA+0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70%) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.