• Journal of Korean Society of Forest Science
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• v.98 no.6
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• pp.719-725
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• 2009

This study was conducted to establish the optimal condition for acclimatization from somatic embryos of Eleutherococcus seoulensis. Torpedo-shaped embryos of Eleutherococcus seoulensis were cultured on 1/3 MS and WPM media supplemented with $GA_3$ (3.0, 5.0 mg/L) for 4 weeks. Plentlets were transferred to 1/2 SH solid medium with 1.0 mg/L $GA_3$ and 0.2% activated charcoal for shoot and root elongation and them elongated plantlets further developed on 1/2 SH medium for 4 weeks. Developed plantlets further elongated into well-shaped leaf and root system on 1/3 SH medium under ventilation condition for 4 weeks. Plantlets grew normally on 1/3 SH basal medium, were acclimated on various soil. Survival frequency of plantlets was influenced by soil type(peatmoss+perlite, perlite, soil on Nam mountain). The highest survival rate to soil was more than 70% when plantlets were 1/3 SH medium under ventilation condition in Nam mountain soil. These results indicate that the systematic procedure of plant production in Eleutherococcus seoulensis could be practically applicable for mass propagation.

• Korean Journal of Medicinal Crop Science
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• v.7 no.4
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• pp.275-281
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• 1999

Embryogenic calli of Codonopsis lanceolata were cultured on MS agar medium containing various concentrations of sucrose as a carbon source. Upon transfer to MS basal medium, somatic embryos of cotyledonary stage converted to plantlets. When sucrose was added with greater than 4%, the number of shoots and roots regenerated from somatic embryo increased. However, the growth of shoots and roots was retarded in agar medium with more than 2% sucrose, but promoted in medium with lower concentration of sucrose. Saponin contents of shoots regenerated from somatic embryos, embryogenic calli, non-embryogenic calli, and native roots were determined by HPLC. Saponin contents of native root was variable, depending on regenerant, embryogenic calli, and cotyledonary embryos. The saponin contents of regenerated roots in medium with high sucrose was similar to native roots. Saponins content based on cell differentiation to shoot and root was dramatically decreased. This results could be effectively controlled for the production of useful secondary metabolites.

• Korean Journal of Plant Resources
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• v.30 no.6
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• pp.654-661
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• 2017

Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.233-239
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• 2003

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.203-209
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• 2005

This study was conducted to improve tocopherol (vitamin E) composition in soybean (Glycine max) by introducing a gamma-tocopherol methyl transferase (${\gamma}$-TMT) gene via Agrobacterium tumefaciens-mediated transformation. Immature cotyledon explants were cocultivated with Agrobacterium tumefaciens. Putative transgenic embryos were selected from immature cotyledons on MS medium supplemented with 40 mg/L 2,4-D containing 100 mg/L kanamycin, 500 mg/L carbenicillin and 250 mg/L cefotaxime. Plantlets were developed from somatic embryos, and then transferred to soil. Nineteen regenerated plantlets obtained on the selection medium from 1,460 cotyledons. However, only 9 plantlets were confirmed as transformed plants. Integration of the transgene into the soybean genomic DNA was confirmed by PCR and Southern blot analysis. HPLC analysis showed that the content of ${\alpha}$-tocopherol in transgenic soybean seeds (AT-1) was approximately 4-fold higher than that of non-transgenic plants. Conclusively, we obtained the transgenic soybean having increased ${\alpha}$-tocopherol content by the overexpression of ${\gamma}$-TMT transgene.

• Korean Journal of Plant Taxonomy
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• v.41 no.2
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• pp.113-118
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• 2011

The somatic chromosome numbers of the Korean species of Senecio L. and two related genera are investigated here. Three different chromosome numbers were found: 2n = 40 in Senecio argunensis Turcz., S. nemorensis L., S. vulgaris L., Tephroseris flammea (DC.) Holub and 2n = 44 in T. phaeantha (Nakai) C. Jeffrey & Y.L. Chen. and 2n = 48 in Sinosenecio koreanus (Kom.) B. Nord., T. kirilowii (Turcz. ex DC.) Holub and T. pierotii (Miq.) Holub. The chromosome numbers of Sinosenecio koreanus (Kom.) B. Nord., Tephroseris flammea (DC.) Holub and T. phaeantha (Nakai) C. Jeffrey & Y.L. Chen. were different from those in previous reports.

• Korean Journal of Plant Taxonomy
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• v.40 no.3
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• pp.153-156
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• 2010

The somatic chromosome number of four Korean species of Leontopodium were investigated. The chromosome number of L. leiolepis (2n = 24) is reported here as for the first time. The chromosome number of L. japonicum (2n = 28) is not varied among the 3 populations on the Korean Peninsula, but that condition is different from the previous reports for Korea (2n = 26) and Japan (2n = 21, 26). L. hallaisanense and L. japonicum, both of which are in sect. Nobilia and similar to each other in gross morphology, have the same chromosome number of 2n = 28. On the other hand the chromosome number of Korean L. leontopodioides (2n = 24) is different from that in Russian reports (2n = 26). The chromosome numbers of all Korean species of the genus Leontopodium could be inferred as tetraploid or aneuploid.

• Plant Biotechnology Reports
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• v.4 no.3
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• pp.237-242
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• 2010

Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard's coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.

• Journal of Plant Biotechnology
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• v.47 no.2
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• pp.164-171
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• 2020

Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

• Proceedings of the Botanical Society of Korea Conference
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• 1993.07a
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• pp.1-36
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• 1993

It is possible to regenerate plants from calli, single cells and protoplasts of numerous species via organogenasis or embryogenesis in cell and tissue culture systems. Also such regeneration of plants can directly occur from cells of explants. However certain plant species has not been yet provided cultures suitable for plant regeneration from cells or tissues. For example, we have to confirm the regenerability of plant from cells before preparing transformed cells for application. Even more, it is very important to notice that regenerated plants in cell and tissue cultures often show structural abnormality. The mojority of those plants is functionally disordered and eventually cases degenerated. One of such examples is vitreous plants which are manifested mainly in the leaves and manifesteds to a lesser extent in the stems and roots. Regenerants in suspension cultures show more frequent vitrification than on gelled media so that relative humidity and water potential are the key factors involved in abnormal morphogenesis in vitro. The other is that somatic embryos formed in media containing BAP or high concentration of sucrose show frequently cotyledon aberrancy such as polycotyledon and born type cotyledon. The embryos with aberrant cotyledon of Codonopsis lanceolata could not germinate or regenerate into plants in many cases. In contrast, the polycotyledon embryos of Aralia cordata germinated in higher percentage than two cotyledonary embryos, but horn type cotyledonary embryos rarely germinated. The major cause of poor germination is the abnormal development of plumule apex meristem.