• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.251-257
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• 1999

Adventitious buds were produced from the cultures of mature zygotic embryos of Larix leptolepis with the highest frequency of 91.7% in SH medium containing 1.0 mg/L zeatin. The most effective cytokinin combination for inducing adventitious buds was 1.0 mg/L zeatin + 1.0 mg/L thidiazuron (40.3%). The highest mean number of adventitious buds induced in 1.0 mg/L zeatin + 1.0 mg/L 2iP or 1.0 mg/L zeatin + 1.0 mg/L kinetin combined treatments. Elongation of the adventitious buds occurred the best on half strength LM salts medium, on which the buds elongated upto 27 mm. Also, the supplement of activated charcoal in medium suppressed the elongation of the adventitious buds. The highest frequency (23.3%) of rooting from elongated shoots was obtained on the medium containing 5.0 mg/L IBA and 0.2 mg/L NAA. The highest number (3.5) of roots was induced on the medium containing 5.0 mg/L IBA alone.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.38-48
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• 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 Plant Resources
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• v.36 no.5
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• pp.527-539
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• 2023

Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer's preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.

• Korean Journal of Medicinal Crop Science
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• v.7 no.4
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• pp.296-302
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• 1999

This experiment was conducted to study the effect of $CO_2$(0, 2, 500, 5, 000, 10, 000ppm) enrichment by enabling ventilation on micropropagation of multi-shoot and on the saponin contents in vitro in Korean ginseng (Panax ginseng C. A. Meyer). Embryo was cultured in Murashige and Skoog medium added 3mg/ l of Indolbutyric acid, Benzyladenin and Gibberellic acid $(GA_3)$, respectively. $CO_2$, enrichment had little effects on the number of adventitious buds and shoots originated from adventitious buds. The ratio of differentiated shoots to adventitious buds were about 50% in $CO_2$, enrichment treatment. The shoots originated from adventitious bud showed more rapid growth and had larger leaf area than the shoots originated from the leaf primordia did. The number of shoot primordia was the highest in 2, 500ppm of $CO_2$ enrichment treatment. On the contrary, 10,000ppm of $CO_2$, enrichment made smaller the number of shoot primordia and ratio of shoots to shoot primordia. The range of shoots differentiated was from shoot primordia were 15. 4 to 23. 9. The rate of dry weight of cultured shoots showed lowest (7. 5%) in control and highest (8. 59%) in 2, 500ppm of $CO_2$, enrichment. Rate of in vitro flower in control was 7.6% and that in 2500ppm of $CO_2$ was about twice (15.7-16.3%) as much as in control. Flower number per a embryo cultured was about 1.2-1.3. In the multi-shoots with callus enriched by 2, 500ppm of $CO_2$, the contents of crude saponin and ginsenosides in multi-shoots alone were higher than in multi-shoots with callus. The characteristics of ginsenosides in multi-shoots were especially the higher content of ginsenoside Rd, Re, and $Rg_1$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.470-486
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• 1995

Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

• Korean Journal of Ichthyology
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• v.18 no.3
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• pp.202-208
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• 2006

An experiment was conducted to investigate the effects of three water temperatures (15, 20 and $25^{\circ}C$) in combination with three salinities (0, 15 and 30 psu) on the oxygen consumption rate of juvenile spotted sea bass, Lateolabrax maculatus (mean body weight $5.5{\pm}0.3g$). The oxygen consumption rates of L. maculatus were measured in triplicate for 24 hours using a continuous flow-through respirometer. Water temperature resulted in significant differences in the mean oxygen consumption rate of L. maculatus (p<0.001), but salinity and combinations of salinity and water temperature did not have (p>0.05). The oxygen consumption increased with increasing water temperatures in all experimental salinity regimes (p<001). Mean oxygen consumption rates at 15, 20 and $25^{\circ}C$ ranged 328.8~342.3, 433.9~441.0 and 651.5~659.9 mg $O_2\;kg^{-1}\;h^{-1}$, respectively. $Q_{10}$ values did not vary with salinity, bud varied with water temperature. $Q_{10}$ values ranged 1.63~1.75 between 15 and $20^{\circ}C$, 2.24~2.26 between 20 and $25^{\circ}C$, and 1.92~1.98 over the full temperature range. The energy loss by metabolic cost increased with increasing water temperatures in all experimental salinity regimes (p<0.001) Mean energy loss rates at 15, 20 and $25^{\circ}C$ ranged 224.6~233.8, 296.3~301.2 and $444.9{\sim}450.7kJ\;kg^{-1}\;d^{-1}$, respectively. These data suggest that the culture of juvenile spotted sea bass is possible without energy loss by salinity difference in freshwater as well as seawater after salinity acclimation. Thus, this result has an application for culture management and bioenergetic model for growth of this species.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.74-81
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• 2022

Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F₁ developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.

• Journal of The Korean Society of Grassland and Forage Science
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• v.23 no.4
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• pp.229-236
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• 2003

This pot experiment was conducted in order to observe the effects of application of combined micronutrients(T$_1$: control. T$_2$; Fe, T$_3$; Fe+Mn, T$_4$； Fe＋Mn＋Cu, T$_{5}$; Fe+Mn+Cu＋Zn, T$_{6}$；Fe+Mn+Cu＋Zn+Mo, T$_{7}$； Fe+Mn+Cu+Zn+Mo+B) on forage performance of pure and mixed cultures of orchardgrass and white clover. The first part was concerned with the changes in the growth, summer depression, and flowering of forage plants. The results obtained are summarized as follows: 1. The T$_3$and T$_{6}$ resulted in a wide chlorosis induced by the Fe-deficiency on orchardgrass. Chlorosis was significantly reduced by the T$_4$and T$_{5}$, whereas the T$_{7}$, T$_2$, and T$_1$showed normal growth without chlorosis symptoms. The T$_{7}$ resulted in the best growth of orchardgrass both in the pure and mixed swards. There was no chlorosis symptom on white clover. whereas the T$_1$, T$_2$and T$_{7}$ showed a relatively good growth with deep green leaf-colour compared with the other treatments in the pure culture. 2. In summer, a summer depression occurred in orchardgrass: this was significantly reduced by the T$_1$and T$_{7}$. The treatments with chlorosis symptoms in orchardgrass partly induced a lodging. Summer depression in white clover did not occur. 3. In the pure culture, the T$_{7}$ and T$_2$in white clover resulted in many flowers and flower-buds compared with the other treatments. The T$_{7}$, especially, showed a long blooming period and an early full bloom compared with the other treatments, whereas the T$_{6}$ and is showed inferior numbers of them. Only the T$_{7}$ resulted in more flowers than flower-buds, and above 1 in the flower/flower-bud ratio except the T$_{6}$ in the mixed culture. 4. It was recognized that the chlorosis of Fe-deficiency occurred not only from the unsuitable ratios of Fe/Mn and Fe/Mo but also from the unsuitable mutual ratios among the cations(Fe, Mn, Cu and Mn), between the anions(Mo and B), and their total ion concentration. It was observed the multiple interactions of Fe${\times}$Mn${\times}$Mo${\times}$B, and the distinct role of B as a regulator.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.420-428
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• 2015

The majority of cultivated varieties of grape have perfect flowers that are clustered in an individual inflorescence. Grape flower has a single pistil, five stamens, a protective flower cap (calyptra), and a calyx. After fertilization, an individual flower develops into a single berry. Although there are a number of reported studies focusing on berry formation, berry enlargement, and sugar accumulation in grape, the morphological studies of flower, including gametophyte morphogenesis and structural change in floral organs, have not yet been studied in detail. In this study, we investigated the flower structure and development characteristics of grape using microscopy and defined the floral development stages 9 to 13 based on microspore or male gametophyte development stage from tetrad to mature pollen. We used seeded diploid table grapes 'Campbell Early' (Vitis labruscana) and 'Tamnara' (V. spp.) as plant materials. At floral development stage 9, pollen mother cells develop to tetrads. During floral development stages 10 to 11, unicellular microspore develop to mid bicellular pollen. At the end of floral stage 12, male gametophyte develops to mature tricelluar pollen. In floral stage 13, the flower cap falls off and flower bud opens. During floral development stages 9 to 12, there were no major changes in calyx length, whereas the length of the flower cap continuously increased. The flower cap-to-calyx length ratio was 2.0, 3.0, 4.5, and 6.5 at floral stages 9, 10, 11, and 12, respectively. The flower cap-to-calyx length ratio was consistent in the two grape cultivars, suggesting that the ratio is a morphological character representing floral development stage. This study provides a reference for determining floral development stage of the two grape cultivars. It will be useful for the determination of optimum time for microspore culture needed to generate doubled haploid lines and appropriate gibberellic acid treatment needed to induce parthenocarpic fruit development in 'Tamnara' grape.

• Journal of Korean Society of Forest Science
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• v.82 no.1
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• pp.26-33
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• 1993

In vitro shoot proliferation and rooting were tested for 2-0 seedlings of half-sib families of 4 plus oaks trees. Nodal segments having axillary buds from 37 families(16 of Quercus acutissima, 10 of Q. variabilis, 7 of Q. serrata, and 4 of Q. mongolica) were cultured on WPM(Woody Plant Medium) supplemented with 0.5 mg/l BA (6-benzyladenine) and 0.01 mg/l NAA(${\alpha}$-naphthalene acetic acid) and subcultured at 2-3 weeks of intervals fur 6 months. In vitro rooting was carried out on GD(Gresshoff and Doy) medium supplemented with 0.5mg/l IBA(indole butyric acid). The capacity for shoot proliferation and rooting was highly varied with families. Generally, white oaks(Q. serrata and Q. mongolica) showed poor response than black oaks(Q. acutissima and Q, variabilis) in shoot proliferation and rooting. Among the total of 37 families, 7 of Q. acutissima, each 2 of Q. variabilis, Q. serrata, and Q. mongolica revealed abilities for continuous shoot proliferation, and the others failed to proliferate. Rooting of the selected oak trees also greatly varied among the families. In Q. acutissima, rooting ratio ranged from 10.0%(CB 25. KG 4) to 89.8%(CB 18). Although 26.7% of KG 16 in Q. variabilis, 3.3% of JN 15 in Q. serrata were rooted, Q. mongolica was not rooted at all in this experimental conditions. No relationship between shoot growth and the rooting ability was observed. Present results suggest the possibility of large-scale micropropagation, but further studies on family differences, shoot-tip necrosis, and callusing of rooting junction are still required to develop reliable micropropagation systems.