• Title/Summary/Keyword: somatic embryogenic callus

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Developing a mass propagation technique for Aralia elata via somatic embryogenesis

  • Moon, H.K.;Lee, J.S.;Kim, T.S.
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2000.10b
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    • pp.16-17
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    • 2000
  • Aralia elata is found in mountain areas all over Korean peninsula. Aralia elata is the scientific name for Japanese angelica tree. The tree belongs to the family Araliaceae, commonly known as ginseng family. Bud sprouts from apical shoot tip of the plants are rich in flavor and thus mainly used for both folk medicine and vegetable. The stalks with apical buds are gathered in the early spring and planted in sandy soil or water in the greenhouse. The sprouting buds are then collected and sold as fresh vegetable. Although the plants have been used for food, they have been cultivated in a very small scale. In spring, local farmers just go around mountain areas to search the trees and gather the stalks as much as they get and sell them to the market. No conservation efforts have been made to stop the exploitation or to save the dwindling population. We tried to provide local farmers with the plants that may be used as an alternative to stalks from wild populations. This will hel! p conserve the wild populations. However, it is hard to propagate them either by conventional cuttings or by seed germination in a short period of time. Mass propagation using tissue culture systems have shown a great promise with several woody plants. Recently we developed a mass propagation technique via somatic embryogenesis system using mature and/ or juvenile explants for Aralia elata. Several factors affecting somatic embryogenesis system including SE(somatic embryo) induction, embryogenic callus proliferation, SE germination, plant regeneration and transplanting to field will be presented. And some problems arising for the somatic embryogenesis system will be also discussed.lso discussed.

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Establishment of Suspension Culture System to Induce Somatic Embryo in Oplopanax elatus Nakai (현탁배양을 통한 땃두릅나무의 체세포배 유도)

  • Kim, Hee Young;Seong, Eun Soo;Lee, Jae Geun;Yoo, Ji Hye;Hwang, In Seong;Kim, Myong Jo;Lim, Jung Dae;Kim, Na Young;Yu, Chang Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.461-465
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    • 2012
  • This study was carried out to select the appropriate medium (especially, carbon and nitrogen source, potassium phosphate and pH) for somatic embryogenesis in order to develop the rapid mass production system in suspension culture of Oplopanax elatus Nakai. Direct somatic embryos were obtained from root explants in the hormone free suspension culture (MS). Combination of $NH_4NO_3$ and $KNO_3$ at the ratio of 1650 (mg/${\ell}$) : 1900 (mg/${\ell}$) obtained the better result to produce somatic embryo in suspension culture. MS medium supplemented with $170mg/{\ell}$ $KH_2PO_4$. The addition of 1 and 3% sucrose was effective for formation of embryogenic callus. Therefore, this report will be helped to improve the establishment for suspension culture in Oplopanax elatus Nakai.

Expression of Porcine Epidemic Diarrhea Virus Spike Gene in Transgenic Carrot Plants

  • Kim, Young-Sook;Kwon, Tae-Ho;Yang, Moon-Sik
    • Plant Resources
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    • v.6 no.2
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    • pp.108-113
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    • 2003
  • This study was carried out to obtain basic information for possibility of oral vaccine in carrot using Agrobacteruim -mediated transformation system. The epitope region of porcine epidemic diarrhea virus (PEDV) spike gene which is classified as a member of the Coronaviridae and causes an acute enteritis in pigs was successfully expressed in carrot (Daucus carota) using the Agrobacterium-mediated transformation system. Hypocotyl segments of in vitro germinated plantlets were infected with Agrobacteriun tumefaciens LBA 4404 harboring PEDV spike gene. Embryogenic callus (EC) was induced on MS selection medium with 1 mg/L 2,4-D, 50 mg/L kanamycin and 300 mg/L cefotaxime after 45 days of culture. Subcultured ECs on MS selection medium without 2,4-D were converted to somatic embryos (SE) of various stage; globular, heart and torpedo stage. Putative transgenic embryos were selected on MS medium with 50 mg/L kanamycin and 300 mg/L cefotaxime. Regenerated plantlets from transformed SE were induced on MS medium containing 50 mg/L kanamycin after 30 days of culture. Genomic PCR confirmed the integration of PEDV spike gene into nuclear genome of carrot and northern blot analysis demonstrated the expression of PEDV spike gene in transgenic carrot.

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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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Somatic Embryogenesis and Organogenesis via Callus Culture from Hydrocotyl maritima Honda (선피막이의 캘러스 배양을 통한 체세포 배발생과 기관분화)

  • Kim, Ok-Tae;Kim, Min-Young;Kim, Kwang-Soo;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.336-339
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    • 2003
  • Hydrocotyle maritima Honda used as medicinal plants for a hemostatic agent was investigated for in vitro regeneration. The petiole explants of H. maritima were cultured on callus induction medium containing growth regulators ($0{\sim}5\;mg/l$, NAA and $0{\sim}5\;mg/l$ 2,4-D) either in single or in combination with $0.1{\sim}2\;mg/l$ BA for 6 weeks. Although single treatments of 2,4-D or NAA resulted in callus formation, the best results were combination of $0.5\;mg/l$, 2,4-D and $0.5\;mg/l$, BA. $5\;mg/l$, NAA and $0.5\;mg/l$, BA, respectively. The highest number of shoot (12 shoots per callus) was achieved with $3\;mg/l$, Kinetin. Also, when pieces of embryogenic callus induced on the medium supplemented with $0.5\;mg/l$, 2,4-D and $0.5\;mg/l$, BA were subcultured on hormone-free medium, somatic embryos were differentiated and developed further into welldeveloped plants.

High frequency plant regeneration system for Nymphoides coreana via somatic embryogenesis from zygotic embryo-derived embryogenic cell suspension cultures

  • Oh, Myung-Jin;Na, Hye-Ryun;Choi, Hong-Keun;Liu, Jang Ryol;Kim, Suk-Weon
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.125-128
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    • 2010
  • Culture conditions were established for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Nymphoides coreana. Zygotic embryos formed pale-yellow globular structures and calluses at a frequency of 85.6% when cultured on half-strength Murashige and Skoog (MS) medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. However, the frequency of pale-yellow globular structures and white callus formation decreased slightly with an increasing concentration of 2,4-D up to 10 $mg\;l^{-1}$ with the frequency rate falling to 16.7%. Cell suspension cultures were established from zygotic embryo-derived calluses using half-strength MS medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. Upon plating onto half-strength MS basal medium, over 92.3% of cell aggregates gave rise to numerous somatic embryos and developed into plantlets. Regenerated plantlets were successfully transplanted into potting soil and achieved full growth to an adult plant in a growth chamber. The high frequency plant regeneration system for Nymphoides coreana established in this study will be useful for genetic manipulation and cryopreservation of this species.

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.

Establishment of Plant Regeneration from Apical Meristem of Sweetpotato (고구마 정단분열조직 유래 식물체 재분화 조건 확립)

  • Lee, Joon-Seol;Ahn, Young-Sup;Chung, Mi-Nam;Kim, Hag-Sin;Jeong, Byeong-Choon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.233-236
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    • 2006
  • This study was conducted to investigate somatic embryogenesis capacity using callus derived from bud meristems in sweetpotato. Shoot apical meristem explants $(height:150{\mu}m;base:\;350{\mu}m)$were cultured on MS medium supplemented with 1 mg/L 2/4-D. Embryogenic callus were observed in five cultivars when their shoot apices were cultured on MS medium supplements with 1 mg/L 2,4-D. After 6 weeks of culture, greater than 80% of the survived explants produced embryogenic calli and the calli gave rise to somatic embryos at frequencies of 72% (Yulmi), 60% (Shinhwangmi), 78% (Geonmi), 70% (KoKei 14), 40% (Sinjami). The regenerated plants developed into whole plantlets after they were transferred onto the fresh hormon-free MS medium of 74% (Yulmi), 82% (Shinhwangmi), 86% (Geonmi), 74% (Kokei 14), 41% (Sinjami) respectively.

Effect of picloram and 2,4-D on plant regeneration from mature and immature embryos of moroccan durum wheat varieties

  • Ahansal, Khadija;Aadel, Hanane;Udupa, Sripada Mahabala;Gaboun, Fatima;Abdelwahd, Rabha;Ibriz, Mohammed;Iraqi, Driss
    • Journal of Plant Biotechnology
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    • v.49 no.2
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    • pp.131-138
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    • 2022
  • An efficient genetic transformation protocol is a fundamental requirement for high regeneration capacity from cultivated durum wheat (Triticum durum) varieties. In this study, wereportedtheeffectsoftwoauxins,2,4-dichlorophenoxyaceticacid(2,4-D)and4-amino-3,5,6-trichloropicoli nicacid(picloram), at a concentration of 2 mg/Laloneandincombination on the embryogenic callus and plantlet regeneration of four durum wheat varieties (Amria, Chaoui, Marouane, and Tomouh) using mature embryos (MEs) and immature embryos (ImEs). Significanteffectsofvariety,culturemedium(theauxinused),andvariety-mediuminteraction were observed on the callus weight and plantlet regeneration of both MR and ImE explants. The medium used for callus induction significantly affected plantlet regeneration (p < 0.001). Comparedto2,4-D, picloram led to a higher plantlet regeneration rate in both ME and ImE explants (19.8% and 40.86%, respectively). Plantlet regeneration also varied significantly depending on the variety and medium used. PicloramledtohighplantletregenerationofbothME and ImE explants in all varieties except Tomouh, which showed high plantlet regeneration of ME explants in 2,4-D. A comparison of ME and ImE responses indicated that ImEs are the best explants for high plantlet regeneration in durum wheat. Ourfindingssuggestthatpicloramisthebestauxin and should be used instead of 2,4-D due to its positive effect on increasing plant regeneration of durum wheat ME and ImE explants.

Somatic Embryogenesis and Adventitious Bud Formation from Zygotic Embryo of Boxthone (Lycium chinense Mill.) (구기자나무(Lycium chinense Mill.) 접합자 배로부터 체세포배 및 부정아 발생)

  • Lee, Jae-Dong;Cho, Duck-Yee;Soh, Woong-Young
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.241-246
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    • 2002
  • Somatic embryos or adventitious buds were formed from the segments of zygotic embryo of Lycium chinense Mill. on Murashige and Skoog (MS) medium supplemented with auxins (2,4-D, NAA, IAA) and / or cytokinins (zeatin, kinetin, BAP). Embryogenic callus formed on MS medium containing 0.5 mg/L 2,4-D and then differentiated into somatic embryos without transfer to hormone free medium. On the other hand, adventitious buds were formed on medium with 0.01 mg/L 2,4-D and 0.5 mg/L zeatin. Among various parts of zygotic embryos, the morphogenic potential was higher in the cotyledonary region, and the most organogenic potential was found in cotyledon followed by radicle, hypocotyl, and whole embryo. Histologically, bipolar structure of the heart-shaped embryos were confirmed and in adventitious buds only shoot apical meristems were found to exist.