• Journal of Ginseng Research
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• v.36 no.4
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• pp.442-448
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• 2012

Somatic embryogenesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology such as medicinally important plants. Single embryos develop into normal plantlets with shoots and roots. Therefore, direct single embryogenesis derived from single cells is highly important for normal plant regeneration. Here we demonstrate that the cyclic secondary somatic embryogenesis in Panax ginseng Meyer is a permanent source of embryogenic material that can be used for genetic manipulations. Secondary somatic embryos were originated directly from the primary somatic embryos on hormone-free Murashige and Skoog medium, and proliferated further in a cyclic manner. EM medium (one third of modified MS medium [MS medium containing half amount of NH4NO3 and KNO3] with 2% to 3% sucrose) favored further development of proliferated secondary somatic embryos into plantlets with root system. The plantlets developed into plants with well-developed taproots in half-strength Schenk and Hildebrandt basal medium supplemented with 0.5% activated charcoal.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.243-248
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• 2006

A optimal procedure for plant production via repetitive secondary somatic embryogenesis in Tilia amurensis is described. Somatic embryos were induced directly from the culture of zygotic embryos on medium with 1.0 mg/l 2,4.-D. Repetitive secondary somatic embryos formed on the surface of the cotyledons and hypocotyls except for the radicles when explants of somatic embryos were cultured on medium with 4.0 mg/l 2,4-D. The highest frequency of secondary embryo-genesis was obtained in the cotyledons (90%) and hypocotyls(83.33%) on MS medium with 1.0 mg/L 2,4-D. The average number of secondary embryos per explant was 25.74 in cotyledon and 24.92 in hypocotyl. When the cotyledon and hypocotyl segments from somatic embryos at different developmental stages were cultured on MS medium with 1.0 mg/L 2,4-D, the highest frequency of secondary embryogenesis was obtained from late cotyledonary secondary embryos. Somatic embryos were transferred to MS basal medium and then they germinated within 2 to 4 weeks of culture. Germinated somatic embryos grew normally into plantlets on WPM medium, producing new shoots. The converted plantlets were acclimatized on artificial soil mixture. These results indicate that the repetitive secondary somatic embryogenesis in T amurensis can offer the possibility to use in vitro culture system for the micropropagation.

• 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.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.149-154
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• 2000

Embryogenic suspension cultures were initiated using embryogenic callus from immature inflorescence explants (Aralia cordata Thunb.) cultured on solid MS medium containing 1 mg/L 2,4-D for 8 weeks and then the embryogenic callus was proliferated in liquid MS medium containing 1 mg/L 2,4-D. After sieving the suspensions (pore size 270$\mu$m), embryogenic cells were cultured in liquid MS medium with cytokinins (kinetin, BA, zeatin) for two weeks. When the embryogenic cells were transferred to liquid MS basal medium, primary somatic embryos were developed after 5 weeks of culture. Secondary embryos were developed directly from the primary torpedo and cotyledonary embryos cultured in solid MS basal medium. Frequency of secondary embryogenesis was higher on medium containing 2 mg/L kinetin than the other cytokinins. Plant regeneration was highly recorded by placing secondary cotyledonary embryos induced from primary cotyledonary embryos in MS medium containing 2 mg/L kinetin or 2 mg/L zeatin (25.4% and 28.6%, respectively). The plant regeneration from secordary embryos was prohibited by tertiary embryogenesis.

• Journal of Plant Biotechnology
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• v.48 no.1
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• pp.44-53
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• 2021

The present work aims to study the induction of somatic embryogenesis in cork oak (Quercus suber L.) from immature zygotic embryos and young apical leaves obtained from 2-month-old seedlings through acorn germination on sterilized peat. The immature zygotic embryos were grown for 1 month on the mineral solution of MS in the presence of 4.52 µM 2,4-D and 30 g/L sucrose. They were then transferred to the same mineral solution with no added growth regulators. In the third subculture, yellow somatic embryos, characterized by two voluminous cotyledons, were differentiated from the radicle of the immature zygotic embryos. The induction of somatic embryogenesis in young leaves required a series of transfers on different culture media containing 30 g/L sucrose and 100 mg/L myo-inositol. Secondary or recurrent somatic embryogenesis occurred within the immature somatic embryo radicles after 1 month of culture on growth regulator-free medium containing WPM macronutrients, MS micronutrients, and vitamins.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.157-164
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• 1995

Intact mature zygotic embryos or their excised cotyledons of ginseng, were cultured on media containing various growth regulators such as auxin (2,4D, IAA) and cytokinin(BAP kinetin). In the culture of intact zygotic embryos, auxin inhibited germination but cytokinin did not Somatic embryogenesis occurred only from those of ungerminated embryos. In the culture of cotyledon segment, medium without growth regulators was the most appropriate to somatic embryogenesis. Somatic embryos were produced sporadically over the surfaces of zygotic embryos on medium containing auxin, while on medium without growth regulators, or media containing cytokinin, somatic embryos formed only on the proximal region of cotyledon. on medium containing 2,4-D, somatic embryos originated from multiple cells which comprised epidermal and subepidermal layers of cotyledon, which resulted in poly-somatic embryogenesis. When these somatic embryos were cultured on the same medium, the primary somatic embryos procured secondary embryos, which arose from epidermal or subepidermal single cells.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.87-90
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• 2004

An efficient plant regeneration system in alfalfa (Medicago sativa L.) through somatic embryogenesis was established. Embryogenic callus was obtained by culture of hypocotyl segments on MS medium with 0.02mg $L^{-1}$ IAA and 1.0mg $L^{-1}$ zeatin after 45 days of culture. Embryogenic calli were converted to the somatic embryos when transferred to either MS medium without plant growth regulators (PGRs) or MS medium containing various cytokinin (BA, kinetin and zeatin). Most of the somatic embryos were developed into plantlets on MS medium supplemented with 0.1 mg $L^{-1}$ kinetin. Also, secondary embryos appeared on the surface of primary embryo but they showed abnormal growth. Regenerated plantlets were transplanted to pots containing vermiculite and perlite for further analysis.

• Korean Journal of Medicinal Crop Science
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• v.14 no.4
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• pp.255-260
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• 2006

Culture conditions for high frequency plant regeneration via somatic embryogenesis from embryogenic cell suspension cultures of Hovenia dulcis are described. Germinated somatic embryos were selected for induction of secondary embryogenesis. Friable embryogenic cells were induced directly from somatic embryos when transfer to 1/3 MS solid or liquid medium lacking plant growth regulators. The temperature strongly effected on induction of secondary embryognesis than other conditions in culture. All somatic embryos produced friable embryogenic cell clumps within 10 days when germinated somatic embryos cultured in 1/3 MS medium at $30^{\circ}C$ in suspension culture. No somatic embryos formed from embryogenic cell suspension cultures at $18^{\circ}C$. Numerous somatic embryos were induced and subsequently developed uniformly into germination stage from suspended cell clumps after 4 weeks of culture on $18^{\circ}C$. Plantlets conversion were observed on $18^{\circ}C$ when germinated somatic embryos were transferred to 1/3 MS solid medium without plant growth regulators or supplemented with 0.1-0.5 mg/l benzyladenine.

• Korean Journal of Plant Tissue Culture
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• v.21 no.2
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• pp.85-90
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• 1994

Attempts were made to regenerate plants from petiole explane of Forest Pimpinella barchycarpa via repetitive somatic embryogenesis. Effective induction of somatic emb교ogenesis was achieved on both MS and modified $B_{5}\;(mB_{5})$ media containing BA + 2,4-D or BA + 2,4-D + NAA under light condition (16-h photoperiod/day) cultures. The explants exposed to the ligt produced numerous somatic embryos while those kept under the dark did not form any on the same medium. Somatic embryos at different developmental stages were observed to arise within a individual explants. Plantlets could be regenerated on $mB_{5}$ basal medium or $mB_{5}$ containing 0.1 mg/L NAA Secondary adventive embryos were formed on the surface of the somatic embryos. Therefore, repetitive somatic embryogenesis could be achieved by secondary embryogenesis. Although the treatment of 2,4-D or NAA alone was effective in callus formation and growth, but not in induction of somatic embryogenesis. Some explants, cultured on NAA-containing media in darkness, produced only adventive roots. The embryogenic potential was maintained for two years when subcultured to BA and 2,4-D containing media with 5 weeks inteval. Regenerated plantlets were maintained on $mB_{5}$ or MS basal media for 4 to 6 more weeks and transferred to soil of an artificial mixture for acclimation. Most plantlets (more than 97%) survived, and grew without any deformity.