• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.511-516
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• 2010

The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.52-56
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• 1985

For the fundamental studies of radiation breeding in edible marine algae, the biological effects on conchospores of Porphyra species by gamma-irradiation were examined. Two varieties, Keun-cham-gim (Porphyra tenera Kjell. form tamatsuensis Miura) and Saga No.5, were chosen for this study, and their conchospores after r-irradiation($5.0{\sim}20.0$ KR) were cultured for 50 days. The results obtained were summarized as follows. 1. Gamma-irradiation in less than the dose of 20KR did not affect germination of conchospores, and almost all spores grew into two cells germ in 24 hours after irradiation, but withering germs were gradually increased in number according to higher exposure within 5 days old culture. 2. High irradiation caused the induction of giant cells, abnormal useless growth of hold-fast, lumpish thalli and callus-like lumpy tissues. 3. The liberation of neutral spores from young germs and carpospores from mature thalli were observed on the frond exposed at $10{\sim}20$ KR irradiation. All spores were normal in division and its size. 4. The best irradiation effect on growth of Keun-cham-gim was observed at 10 KR dose, whose growth-rates were $140\%$ in wet weight and $108\%$ in mean frond area, but only $48\%$ was recorded in wet weight at 20 KR exposure. Saga No.5 were in contrast with Keun-cham-gim, and their most growth-rate was $400\%$ in wet weight ($258\%$ in frond area) at 10 KR irradiation and the worst was $20\%$ at the dose of 20 KR. 5. The withering phenomenon to death by treatment of gamma-ray presented substantial difference between two varieties. Survival rate compared with control in Keun-cham-gim was $70.7\%$ at 20 KR, but that in Saga No.5 recorded $47.0\%$ at same dose. 6. Synthesizing the results of high and low r-irradiation, it was suggested tat high r-irradiation in more than 5.0 KR inhibited conspicuously the growth of germs derived from conchospores, and about half of them withered at 15.0 KR dose or more.

• Journal of Korean Society of Forest Science
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• v.87 no.2
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• pp.164-172
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• 1998

This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

• 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 Tissue Culture
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• v.24 no.3
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• pp.129-135
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• 1997

Morphogenetic responses were investigated by culturing embryo and leaf explants of Korean wild type tea plant, Camellia sinensis (L.) O. Kuntze. Induction of direct somatic embryogenesis as well as adventitious and/or axillary shoots was obtained from mature zygotic embryo cultures on Murashige and Skoog (MS) basal medium having 5 to $20\mu\textrm{M}$cytokinin a lone. Morphogenetic response was decreased dramatically by the addition of auxins tested. One hundred percent of induced and isolated shoots formed roots after four weeks of culture on half-strength MS or quarter-strength Schenk and Hildebrandt (SH) media supplemented with $10\mu\textrm{M}$indole-3-butyric acid (IBA). Immature zygotic embryos were shown to be a suitable explant for embryogenic callus formation in the presence of 2, 4-dichlorophenoxyacetic acid(2, 4-D) in basal medium. Mature zygotic embryo originated leaves were used to test their ability for mophogenesis by incorporating plant growth regulators such as IBA, naphthyl-1-acetic acid (NAA), and 6-benzylaminopurine (BAP). Apparently, the morphogenetic responses of the cultured explant sources on the types and/or levels of plant growth regulators tested were observed visually.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.93-97
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• 1999

Shoot tips with 2 leaf primordia were cultured to induce shoot primordia in MS liquid medium supplemented with several concentrations of BA and hIAA under the conditions of 10,000 lux illuminations for 24 h and of vertical shaking of 2 rpm in carrot. Two F$_1$ hybrids and two male sterility lines were used. Shoot primordia were only induced in the medium supplemented with 2.0 mg/L of BA and 0.2 mg/L of NAA. Genotypic specificity and seasonal effect of donor parents on shoot primordia induction were not observed and average 15-20% of the planted dornes developed to shoot primordia. The induced shoot primordia were successfully propagated by subculture in the same medium. However, they were grown into three different types during multiplication, that is, the type with multiple small shoots on the surface, the type of without any shoot, and the type of callus. Shoot primordia clusters with small shoots on the surface differentiated multiple shoots successfully in 1/2 MS solid medium supplemented with 0.2 to 1.0 mg/L of IAA and 0.2 to 1.0 mg/L of kinetin. New shoot primordia with small shoots were well formed when pieces bigger than 2 mm in diameter of the out layer of the shoot primordia cluster with small shoots were subcultured. No differences of multiplication and shooting ability and chromosomal variation of shoot primordia were observed until the 13th sub-culture.

• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.109-115
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• 2000

Effects of the heavy-ion ($^{14}$ N or $^{20}$ Ne) beam irradiation on the response of anthers, growth of calli, germination of seeds, and the early growth after the germination of tobacco (Nicotiana tabacum L. cv. BY-4) were investigated. Anthers precultured for 10 days before the irradiation became brown without callus or shoot induction over 20 Gy of $^{14}$ N and $^{20}$ Ne ion beam irradiation. Relative growth rate of the cultured calli was reduced by the irradiation and became brown significantly 2 weeks after the $^{14}$ N and $^{20}$ Ne ion beam irradiation over 50 Gy. The increased intensity of the heavy-ion ($^{14}$ N, $^{20}$ Ne) beam irradiation resulted in the delay of seed germination and the inhibition of the early growth both in water-treated and non-treated seeds before the irradiation. In addition, the heavy-ion beam irradiation to the imbibed seeds inhibited seed germination more than that to the non-imbibed seeds. The screening approach of non-imbibed seeds with heavy-ion beam irradiation using in vitro culture system was more useful than the filter-paper germination method in investigating the characteristics of heavy-ion beam-irradiated seed population and the screening of morphological variants at the early stage of the plant growth.

• Archives of Plastic Surgery
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• v.34 no.3
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• pp.298-304
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• 2007

Purpose: This study was undertaken to investigate the osteogenic induction potential of PGA & FBS mixture on a calvarial defect in the rabbit. Methods: Twenty New zealand white rabbit, weighing from 3.5-4kg were allocated into each of the three groups. Four 8 mm sized bone defects were made on the parietal bone by drilling. In group I, the bony defects were implanted with $50{\mu}m$ thickness film containing mixture of PGA and FBS. In group II, with PGA only film, & in group III, the bony defects were left with no implants. Results were evaluated by using morphologic change, radiographic study, biochemical study and histologic examination at 1 week (group I n=7, group II n=7, group III n=14), 2 weeks (group I n=6, group II n=6, group III n=12) and 3 weeks (group I n=7, group II n=7, group III n=14) following implantation. Results: In the morphologic & radiographic study, the formation and corticalization of callus were observed earlier in group I than in groups II and III (p < 0.05). In histological examination, group I showed more abundant and faster new bone formation than in group II and III. In biochemical analysis, group I displayed more activity than in group II and III. Group I also showed more abundant osteopontin, osteocalcin than groups II and III. Conclusion: In conclusion, the results demonstrate that the mixture of PGA and FBS has an effect on osteoblastic formation in the rabbit model. It is considered that further evaluation of long term results on resorption, immunologic tissue reaction and response of applied mixture in the human model will be needed.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.197-203
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• 1999

The albino plants of tobacco (Nicotiana tobacum L. cv. BY-4) were isolated from seed populations that were induced by heavy-ion ($^{14}N$) beam irradiation to proembryo and the in vitro growth and differentiation have been characterized. The in vitro cultured albino plants showed significant reduction of chlorophyll content and possessed larger number of stomata on both upper and lower epidermis than that of wild-type plants. Stem growth of the mutants remained dwarfed, however, the internode recovered its normal length after GA$_3$ treatment (10.0mg/L) on the MS medium containing sucrose under continuous light. When explants of leaf blades of albino plants were cultured, multiple shoots formed directly on MS medium containing 1.0mg/L of BAP or kinetin and a large number of calli were induced on the MS medium containing 1.0mg/L NAA or 1.0 mg/L 2,4-D. The albino calli regenerated multiple albino plantlets in the MS medium containing 0.1mg/L NAA + 1.0 mg/L BAP. No significant differences between the wild-type and albino plants were detected in the multiple shoot induction, callus formation from the explants and the plantlets regeneration from calli. In addition, albino plants have a similar organogenesis Pattern to that of the wild-type in the media with different combinations of NAA (0 to 5.0mg/L) and BAP (0 to 5.0mg/L) treatment. These results indicate that the albino mutant has the same normal regeneration ability as that of wild-type, although the mutant has lost functions in photosynthesis, such as pigmentation.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.159-164
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• 2001

To produce novel bioactive flavonoids, Gericudranin A and Gericudranin B, a cell culture system of Cudrania tricuspidata including callus induction and optimization of culture conditions was established. Friable calli were efficiently induced from the hypocotyl segments of seedlings on B5 medium supplemented with 1.0 mg/L NAA, 0.1 mg/L kinetin and 3% sucrose. Several factors were optimized for the Gericudranin production and the cell growth in suspension cultures. Low level of basal salt medium (1/8 MS), 1.0 mg/L IAA and 0.1 mg/L zeatin, and high level of sucrose (5%) were effective for the production of Gericudranins, whereas WPM with 1.0 mg/L NAA, 0.1 mg/L zeatin, and 5% sucrose were more effective for the cell growth. When cells were cultured on MS liquid medium supplemented with 1.0 mg/L IBA, about 2200 $\mu\textrm{g}$/g dry wt of Gericudranin A could be produced. The level might be about 10 times of the native inner bark. About 2350 $\mu\textrm{g}$/g dry wt of Gericudranin B was also produced on MS liquid medium with 5% sucrose, 1.0 mg/L NAA, 0.1 mg/L kinetin. The content was estimated about 3 times of the level of native inner bark.