• Korean Journal of Plant Tissue Culture
• /
• v.25 no.3
• /
• pp.207-217
• /
• 1998

Total of 78 strains were characterized based on the morphological characteristics of colonies isolated on Schroth, and New & Kerr's media for selection of hypervirulent wild-type Agrobacterium spp from galls, hairy root-like process and soil of Populus, Malus, Salix and Diopyros in Korea. Among them, 48 strains were able to induce tumors in carrot disc. Hypervirulent A. tumefaciens SP101 and SM042 were identified as biotype 1 and biotype 2, respectively, These strains formed fast growing, larger tumors as compared to those induced by other strains. The binary vector pGA643 with kanamycin resistant gene was mobilized from E. coli MC100 into A. tumefaciens strain SM042 isolated from soil, and/or disarmed vector PC2760 using a triparental mating method with E. coli HB101/pRK2013, and transconjugants, A. tumefaciens SM643 and PC643 were obtained in minimal media containing kanamycin and tetracycline. Tobacco tissues were cocultivated with conjugant Agrobacterium and then transferred to selective medium with 2,4-D and kanamycin to induce the transformants. Calli were formed more efficiently in cocultivation with A. tumefaciens SM643 than that with A. tumefaciens PC643. Most of calli transformed with A. tumefaciens PC643 were friable and regenerated into normal plantlets, while the calli transformed with A. tumefaciens SM643 were compact, hard, and mixed with friable calli. The friable calli formed normal shoots, while compact calli did not form shoots but only grew to typical compact tumor calli. When the shoots formed directly from tobacco stems without callus induction after transformation by A. tumefaciens SM643 with wild-type Ti-plasmid, normal transformed plants can be induced without using disarmed Ti-plasmid.

• Journal of Ginseng Research
• /
• v.43 no.1
• /
• pp.38-48
• /
• 2019

Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

• Korean Journal of Agricultural Science
• /
• v.7 no.2
• /
• pp.65-76
• /
• 1980

In order to determine the effects of cytokinin and auxin on organ formation from tissue of garlic cloves, leaf blades and basal tissues contained meristem of garlic (Allium sativum L.) cloves harvested in 1979 (old cloves) and 1980 (new cloves) were explanted on a MS medium contained various levels of BA ($N^6$-benzyl amino purine), NAA (naphthalene acetic acid), and 2, 4-D (2, 4-dichlorophenoxyacetic acid). And some of the new cloves were explanted on a media contained BA and NAA after chilling treatment at $4^{\circ}C$ for 10, 20, 30 and 40 days. 1. In a culture of leaf blades of old cloves, shoots were differentiated on a medium supplemented with 2mg/l of BA and NAA. 2. Callus was grown as a quite straw-coloured globular mass on a medium contained 0.2 or 2mg/l 2.4-D. 3. As subcultures of globular calli, shoots and roots were differentiated on a medium contained 2mg/l BA and 0.5 or 1 mg/l NAA, whereas no shoots was shown on a conterol. 4. Shoots were differentiated in a culture of leaf blades of new cloves, but they were not in an old cloves in control, and better effect was shown on a medium contained 2mg/l BA and 1mg/l NAA. However shoots were no differentiated from leaf blades chilled at $4^{\circ}C$ for 30 or 40 days at the same condition. 5. Large numbers of adventitious shoots could be obtained from basal region of garlic cultured on a medium contained 1mg/l BA and 4mg/l NAA, or 2mg/l BA and 2mg/l NAA.

• Korean Journal of Plant Resources
• /
• v.20 no.1
• /
• pp.43-49
• /
• 2007

This study was conducted to examine the pattern and characteristics of flowering and leaf emergence for twenty two kinds of the genus of Lycoris sp. One species of Lycoris was flowered on July to August, twelve species on August, and nine species on September. Shape of flowers were as follows : Nine species belong to L. radiara-shaped flower, seven species L. squamigera-shaped flower, and seven species the intermediate type. Flower color of three kind were an order of descent red and pink. seven kind white, four kind yellow, three kind orange, and two kind purple colors. Ten Lycoris species were good fertile, and two kinds were poor fertile. The average length of pollen grain was ranged from $48.53\;to\;88.62{\mu}m$, and the width from $22.87\;to\;33.67{\mu}m$. Eleven kinds were spring-leaf emergence type, and ten kind autumn-leaf emergence types. Hunter values in leaves were as follows : $L^*$ value was ranged from 31.6 to 45.2; $a^*$ values from -6.74 to -17.46; $b^*$ values from 6.19 to 21.89 Leaf width was ranged from 0.90 to 2.35cm, and its shape was oblong. The length of epidermal cells was 0.33 to 0.75mm, and the width was $38.53\;to\;90.00{\mu}m$. Most of stomata were distributed in the back side of leaves. The length of stomata was ranged from $67.12\;to\;104.89{\mu}m$, and its width was from $14.90\;to\;71.52{\mu}m$.

• Journal of Life Science
• /
• v.16 no.2 s.75
• /
• pp.289-296
• /
• 2006

In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.

• Korean Journal of Plant Tissue Culture
• /
• v.28 no.1
• /
• pp.7-13
• /
• 2001

Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

• Korean Journal of Pharmacognosy
• /
• v.5 no.2
• /
• pp.111-124
• /
• 1974

The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.