• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.370-375
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• 2015

This experiment compared the regeneration conditions of the radiation mutant spray chrysanthemum 'purple ND'. The four different flower blooming stages (S1: 10% opened flower, S2: 30% opened flower, S3: 50% opened flower, and S4: 70% opened flower) and different petal parts (TBOP: the basal of petal and TEOP: the end of petal) were used to compare regeneration conditions between plants grown in MS medium supplemented with IAA and BAP. The highest adventitious shooting rate was identified in plants grown on the IAA $1.0mg{\cdot}L^{-1}$ and BAP $2.0mg{\cdot}L^{-1}$ when using the end of petal at the S2 stage. It displayed 79.2% regeneration and produced 33.4 shoots. Rooted plantlets were successfully established in the greenhouse, showing the same morphological characteristics of vegetative and reproductive organs with those of the mother plant. Flow cytometry analysis revealed no ploidy variation between the regenerated plants and the mother plant grown under greenhouse conditions.

• Korean Journal of Medicinal Crop Science
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• v.12 no.6
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• pp.500-507
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• 2004

The establishment of an efficient protocol of plant regeneration from leaf explant cultures of Perilla spp. is reported. Regenerated shoots were obtained from leaf explant cultures on solid MS medium containing different concentrations of cytokinins and auxin. The effect of cytokonin and auxin differed depending on each acession. The combination treatments of high level of cytokinin and low level of auxin was more effective for plant regeneration in Perilla frutescens. The best concentration of sucrose was 3% for regeneration. Of spermidine, spermin and putrescine. treatments, the most effective treatment for plant regeneration was $10\;mg/{\ell}$ spermidine.

• Journal of The Korean Society of Grassland and Forage Science
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• v.26 no.2
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• pp.69-76
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• 2006

To optimize tissue culture responses for genetic transformation of Kentucky bluegrass, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar 'Newport' as explant tissues. The optimal concentration of 2,4-D (2.4-dichloro phenoxy acetic acid) for the induction of embryogenic callus from mature seed was 3 mg/L. Plant regeneration frequency was 54% when embryogenic callus was cultured on the regeneration medium supplemented with 1 mg/L 2,4-D and 3 mg/L of BA (6-benzyladenine). Addition of 1 g/L of casein hydrolysate and 500 mg/L of L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 60.8% and 58.3%, respectively. Regenerated plants were grown normally when shoots transplanted to the soil. A rapid and efficient plant regeneration system established in this study. We suggest that the results may be useful for molecular breeding of Kentucky bluegrass through genetic transformation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.28 no.3
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• pp.165-170
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• 2008

Timothy (Phleum pratense L.) is an important grass species as forage. In order to optimize tissue culture conditions of timothy, the effects of plant growth regulators on callus induction and plant regeneration was investigated with mature seeds of colt cultivar. The optimal concentration of 2,4-D for the induction of primary callus from mature seeds was 3 mg/L. The highest embryogenic callus frequenc (25%) was observed when the mature seed were cultured on MS medium supplemented with 3 mg/L 2,4-D and 0.1 mg/L BA. The highest plant regeneration frequency was observed when type B callus was transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots were transplanted to the soil. A short tissue culture period and regeneration system would be beneficial for molecular breeding of timothy by the production of transgenic plant.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.80-80
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• 2018

Hosta is a genus of the family Asparagaceae and distributed in East Asia. There are six Hosta species (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) native to Korea and among them, four species (H. minor, H. jonesii, H. venusta and H. yingeri) are endemic to the Korea peninsula. Hosta is generally propagated by seed, crown division or tissue culture. However, tissue culture is a more efficient method to mass proliferation, a new cultivar development and disease-free plantlet production in a limit time. Hence, we conducted this study to evaluate the influence of various plant growth regulators (PGRs) treatments on the induction of callus and somatic embryo of the six Hosta species. Leaf, petiole and root were used to select optimum tissue culture explants. Petiole explants thus only were used for callus induction and somatic embryogenesis with TDZ (0.1, 0.5 or 1.0mg/L) and NAA (0.1 or 0.5 mg/L) combinations. After 12 weeks of culture, the highest rate of somatic embryogenesis was achieved on modificated MS medium containing 1.0 mg/L TDZ and 0.1 mg/L NAA in H. capitata and H. minor (15.5%, respectively), 0.1 or 0.5 mg/L TDZ and 0.1 mg/L NAA in H. jonesii (22.2%), 1.0 mg/L TDZ and 0.5 mg/L NAA in H. yingeri (26.7%), and 0.1 mg/L TDZ and 0.5 mg/L NAA in H. venusta (53.3%). H. clausa showed very low effect on somatic embryogenesis by PGRs; 2.2%. There was interspecies difference to PGRs respond for callus and somatic embryo induction. Regenerated multiple shoots and plantlet of H. minor, H. jonesii, H. venusta and H. yingeri were obtained via somatic embryogenesis.

• 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.

• Journal of The Korean Society of Grassland and Forage Science
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• v.13 no.3
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• pp.177-183
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• 1993

This study was conducted to obtain the transformed tobacco plants with S. cerevisiae Acid phosphatase gene(PH05) using Agrobacterium tumefaciens and th confirm plant transformation and gene expression. the results obtained were summarized as follows: APase activity of Saccharomyces cereviase NA 87-11A was remarkably showed up as deep red color when assayed by Tohe and Oshima(1974). PH05 fragment, Apase gene, was obtained from pVC727G and the graphically estimated size was about 1.5kb by agarose gel electrophoresis. The sequencing results of 5'end and 3'end of PH05 using dideoxy chain termination method were coinsided with the full length nucleotide already. pBKJ I vector was constructed by isolation of PH05 fragment from pVC727-1 and pBKSI-1 digesred with Sma I and Xba I. Isolated plasmid from transformed A. tumefaciens with constructed pBKJ I when it was electrophoresed with agarose gel. The dosc of tobacco leaf was cocultivated 재소 transformed Agronacterium tumefaciens. Transformed shoots were selected on kanamtcin-containing MS-n/B medium and they were regenerated. The transgenic tobacco plants were elucidated by isolation of genomic DNA and genomic southern hybridization using ${\alpha}-^{32}P$ labelled PH05 fragments. The PH05 in transformed tobacco plants was expressed in leaf, stem and root, and its APase activity was estimated as deep red color by Tohe method.

• Korean Journal of Weed Science
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• v.17 no.4
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• pp.390-399
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• 1997

This experiment was conducted to introduce PAT (phosphinothricin acetyltransferase, non-selective herbicide bialaphos resistant gene) gene into potato (Solanum tuberosum. cv. Desiree). Optimal shoot regeneration from leaf discs and stem segments was obtained in MS medium supplemented with 0.1 mg/L IBA and 0.5 mg/L BA, and the frequency of shoot regeneration was 54% in left discs and 46% in stem segments. In this condition, leaf discs and stem segments of potato were co-cultivated with A. tumefaciens MP90 which contained binary vector with GUS: :NPTII gene and PAT gene. Transgenic shoots were regenerated from leaf and stem-derived calli on selection medium with 100mg/L kanamycin. The 100${\mu}M$ acetosyringone treatment during the co-cultivation highly enhanced(4 times than the control) the shoot regeneration on selection medium. When the putative transgenic plants were transferred to medium with 10mg/L basta, all of them were survived. After PCR. GUS test, and Southern blot analysis of the survived plant, we confirmed that the gene was stably integrated into the potato genome and expressed. After the transgenic plants were transplanted in soil, and the transgenic plants were sprayed with the herbicide basta (300ml/10a), the transgenic plants remained green but control plants were died.

• Journal of The Korean Society of Grassland and Forage Science
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• v.24 no.1
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• pp.21-24
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• 2004

To determine lethal temperature of alfalfa (Medicago sativa L. cv Vernal) at heat-stressed conditions, seedlings grown in a small pots for 4 weeks were subjected to different temperature regimes of heat treatment. No apparent demage was observed when the plants were treated at 45, 50 or $60^{\circ}C$ for 1 h. Heat treatments at 60 and $65^{\circ}C$ for 1 h, several plants were withered and showed damage symptom on their leaves. When the plants were exposed to $70^{\circ}C$ for 1 h, most of leaves were severely withered, but it was not lethal conditions for the whole plants. By contrast, most of plants were died within one day after heat treatment at $80^{\circ}C$ for 1h. Furthermore, plants exposed to $80^{\circ}C$ for 50 min were also died within 7 days. It was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 45 min. These results indicate that heat treatment at $80^{\circ}C$ for 50 min is an optimum condition to distinguish the lethality of alfalfa plants. Simple viability assay system established in this study will be useful fer selection and characterization of heat-tolerant transgenic alfalfa plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.24 no.1
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• pp.25-28
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• 2004

To determine lethal temperature of orchardgrass (Dactylis glomerata L. cv. Janbeol 102) developed in Korea at heat-stressed conditions, seedlings grown in a amall pots for 4 weeks were treated at $45^{\circ}C$, $50^{\circ}C$ or $55^{\circ}C$ for 1 h. Heat treatments at $60^{\circ}C$ and $65^{\circ}C$ for 1 h, several plants were withered and showed damage symptom on their leaves. When the plants were exposed to $70^{\circ}C$ for 1 h, most of leaves were severely withered, but it was not lethal conditions for the whole plants. By contrast, most of plants were died within one day after heat treatment at $80^{\circ}C$ for 1h. Furthermore, plants exposed to $80^{\circ}C$ for 55 min were also died within 7 days. It was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 50 min. These results indicate that heat treatment at $80^{\circ}C$ for 55 min is an optimum condition to distinguish the lethality of orchardgrass plants. Simple viability assay system established in this study will be useful for selection and characterization of heat-tolerant transgenic orchardgrass plants.