• Journal of Forest and Environmental Science
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• v.30 no.4
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• pp.405-411
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• 2014

Wild Korean ginseng has been recognized as highly precious medicine since ancient times. Nowadays, the population of wild ginseng in the forest of Korean peninsula is very rare due to indiscreet harvest. In this work, we investigated the plant regeneration via somatic embryogenesis from embryogenic callus of old wild ginseng (more than 50 years-old) and compared the features of plants regenerated from 5-years old and 50 years-old ginseng. Induction of embryogenic callus from adventitious roots of 50 year-old wild ginseng required 83 weeks of culture, but only 10 weeks were sufficient for 5 year-old ginseng. Height and width of plants derived from the old wild ginseng was smaller and slender compared to the plantlets derived from 5 year-old ginseng. Total chlorophyll contents was 2-6 time lower in plantlets regenerated from 50 year-old wild ginseng than those from 5 year-old ginseng, but anthocyanin content was higher in 50 year-old ginseng. Our results revealed that plants regenerated from old wild ginseng have different morphological and physiological characters probably due to age-dependent phenomenon.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.237-242
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• 2006

Commercial cultivars and elite germplasms of barely (Hordeum vulgare L) are still recalcitrant to genetic transformation because of the lack of an efficient regeneration system. In this study, we established an efficient plant regeneration procedure from embryogenic calli derived from mature embryos. Callus induction from germinated mature embryos was best as over 95% in CIM medium (CI medium containing $2.5mg/{\ell}$ dicamba) under dark incubation. Development of embryogenic callus was highest as over 50% in CI3D medium (EC medium supplemented with $3mg/{\ell}$ 2,4-D). The highest regeneration of plants from embryogenic callus (40%) was obtained with CIS medium ($SI+1mg/{\ell}IAA\;and\;2mg/{\ell}\;BA$). These plant regeneration conditions could be useful in improving barley transformation efficiency.

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.15-19
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• 1999

For the induction of somatic embryogenic callus, the penduncle explants of Corydalis remota for peatinata were cultured on MS basal media supplemented with 2,4-D, kinetin and zeatin. The highest embryogenic callus formation was observed on the media containing 2.0 mg/L of 2,4-D and 2.0 mg/L of zeatin. The somatic embryogenesis on the media with 0.5 mg/L of cytokinin (zeatin or kinetin) were excellent under light condition, however somatic embryos abnormally developed into plantlets. Normal dicotyledonary plantlets were found on MS medium supplemented with 1.0 mg/L of zeatin. When MS medium with 2,4-D plus cytokinin and with BAP were used, the secondary somatic embryogenesis took place in root explants of the regenerants derived from in vitro somatic embryogenic callus.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.133-140
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• 2008

The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and $GeneFishig^{TM}$ DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.

• KSBB Journal
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• v.6 no.4
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• pp.385-388
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• 1991

Embryogenic cell suspension cultures of B. striata were established as transferred selected embryogenic callus into liquid medium. Protoplasts isolated from embryogenic cell suspensions were electroporated in buffered solutions containing plasmid DNA of pBI121. Transient GUS (beta-glucuronidase) activity measurement and selection for kanamycin resistent showed that expression of foreign genes and stable transformation were achieved. GUS transient gene expression was increased by increasing DNA concentration of pBI121 plasmid and affected by the level of the applied voltage. An optimal level of GUS activity was obtained after electroporation with a pulse of 200-300 voltage/1180 uF. Protoplast viability was up to the 80% at the optimal voltage.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.5
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• pp.356-360
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• 2005

Mature embryos were aseptically excised with a scalpel and sliced in fragments measuring 0.5 mm in diameter (sliced mature embryo fragment; 4 ${\~}$ 5 fragments/one embryo). Sliced mature embryo fragments of six wheat cultivars were cultured to develop an efficient method of callus induction and plant regeneration. Callus derived from sliced mature embryo fragments showed a good capacity to embryogenesis and regeneration. Furthermore sliced mature embryo fragments decreased contamination from fungi and bacteria. The high efficiency of callus induction were obtained Keumkangmil and Bob­white. For plant regeneration, selected embryogenic calli were transferred to two types regeneration media. An average number of green spots per callus was 4 to 5 in regeneration media after about one week. Percentage of plant regeneration showed high in regeneration medium containing 0.1 mg/l 2,4-D and 5 mg/l zeatin. Especially, Keumkangmil ($27.5\%$) and Bobwhite ($33.3\%$) showed high regeneration efficiency. This regeneration system from sliced mature embryo fragments may provide an effective and convenient explant for plant transformation studies.

• Journal of Korean Society of Forest Science
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• v.94 no.1 s.158
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• pp.39-44
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• 2005

An effective micropropagation system for Liriodendron tulipifera L via somatic embryogenesis was established using immature seeds. Immature seeds from five individual trees were bisected longitudinally and cultured on two basal media (MS and B5) containing different combinations of 2,4-D and TDZ to induce callus and embryogenic tissue under light ($40{\mu}mol\;m^{-2}s^{-1}$, 16 hr/day) or complete darkness at $25{\pm}2^{\circ}C$. There was no distinctive difference on callus and embryogenic tissue induction between the two basal media with PGRs. Optimum culture medium appeared to be MS medium supplemented with 1.0mg/L 2,4-D and 0.01mg/L TDZ plus 3% sucrose. Nonembryogenic callus induction rate was not significantly different among the genotypes. However, However, the embryogenic callus induction frequency differed greatly by the genotypes ranging from 55% to 72% when cultured in the dark. Generally, the cultures maintained in the dark tended to show normal somatic embryo development as well as embryogenic tissue formation and this was confirmed by histological examination. Above results suggest that a proper selection of mother tree and dark culture condition are necessary to optimize somatic embryogenesis system of Liriodendron tulipifera.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.49-55
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• 2017

We investigated the callus induction and plant regeneration ability of 16 maize genotypes, including the Korean inbred lines, using 9 to 15 day-old immature zygotic embryos from maize grown in pots and from field cultures. Immature zygotic embryos placed on MS medium supplemented with L-proline 0.7 g/L, MES 0.5 g/L, Dicamba 1.5 mg/L, 2,4-D 0.5 mg/L, $AgNO_3$ 4 mg/L, and sucrose 20 g/L, showed the highest frequency of callus induction. The highest number of shoots regenerated when the embryogenic callus were transferred to MS medium supplemented with 5 mg/L zeatin. The root formation was observed when shoots were grown on MS medium supplemented with 0.2 mg/L indole-3-butyric acid (IBA). Additionally, under the same culture conditions, immature zygotic embryos from maize grown in the field also had a high frequency of plant regeneration. Except one genotype, 15 genotypes showed callus induction and shoot regeneration. Among the 16 genotypes tested, H99, B98, HW3, and B73 yielded the best plant regeneration. H99 showed maximum shoot formation from the primary embryogenic callus. The results suggest that genotypes and growth conditions of the maize plant plays very important roles for enhancing the embryogenesis competence of immature zygotic embryos. The successful regeneration from immature zygotic embryos of maize inbred lines provides a basis for molecular breeding of new cultivars by genetic transformation.

• Journal of Plant Biotechnology
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• v.5 no.3
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• pp.169-172
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• 2003

Somatic embryogenesis was initiated from in vitro grown leaf explants of rose following an induction period of four weeks on MS basal medium supplemented with auxin and several subcultures on MS medium with cytokinin. '4th of July' showed the highest regeneration frequencies on 1 mg/L NAA followed by culture on medium with 4 mg/L zeatin. The embryogenic callus was propagated on MS medium with NAA, zeatin and $GA_3$. Germination of somatic embryos was achieved on MS medium with 1 mg/L BA. Somatic embryo derived plantlets were hardened and successfully transferred to the greenhouse.

• Journal of Plant Biology
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• v.33 no.4
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• pp.271-276
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• 1990

Our study was carried out for plant regeneration via somatic embryogenesis from immature seeds of Bletilla striata. The highest frequency of embryogenic callus formation was obtained from the immature seeds (at 150 days after pollination) cultured on Hyponex and VW medium supplemented with 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg/l kinetin under the dark condition. Multiple somatic embryos were induced when embryogenic callus was transferred to VW medium without growth regulators under continued illumination. Somatic embryos were observed histologically with scanning electron microscopy. Regeneration of Bletilla striata was obtained from somatic embryos with a well-defined scutellum and coleoptile as well as with one or more shoot primordia and root primordia. We think that these methods for orchid multiplication must be useful to access clonal propagation of orchids.