• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.77-83
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• 2010

Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

• Korean Journal of Medicinal Crop Science
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• v.5 no.1
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• pp.21-27
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• 1997

This experiments were conducted to determine the effect of thidiazuron on forming tuberlets and plant regeneration of Pinellia ternata T. by tissue culture. The addition of $5\;{\mu}M$ TDZ to the medium had better regeneration than that of any other treatments of NAA and TDZ. At the combination treatments of NAA and TDZ, as the level of thidiazuron increased, the rate of shoot regeneration was incresed while the increment of NAA concentration inhibited the rate of shoot regeneration. The supplement of $5\;{\mu}M$ thidiazuron produced the best number of micro-tubers per explant and the number of micro-tuber formed was 25 in MS medium and 29 in MG medium on 30 day culture, respectively. Microtuber formation was the best on MG medium with 1.0 mg/l NAA and $5\;{\mu}M$ thidiazuron. MG medium was superior to MS and B5 medium for the growth of tuberlets. Half strength of MS medium with NAA 2 mg/l was the most effective for root formation. Rooting ability on nursery soil of plantlets produced in in vjtro was good as a 80% after 3 weeks.

• Journal of The Korean Society of Grassland and Forage Science
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• v.29 no.4
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• pp.291-298
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• 2009

In order to optimize tissue culture conditions for genetic transformation of Miscanthus sinensis, we investigated the effects of different plant growth regulators on callus induction and plant regeneration using mature seeds as explant. Dehusked mature seeds were cultured on MS medium supplemented with 3 to 10 mg/L 2,4-D, dicamba or NAA, 30 g/L sucrose and 750 mg/L $MgCl_2{\cdot}6H_2O$. A number of combinations of auxin and cytokinin (BA, kinetin) were also used. MS medium containing 3 mg/L 2,4-D was found optimal for embryogenic callus induction (75.7%) from mature seed. The highest number of plants were regenerated (44.6%) upon transferring the embryogenic callus to MS medium supplemented with 1 mg/L 2,4-D plus 2 mg/L BA. This high efficient plant regeneration system could be useful to use for molecular breeding of new cultivars by genetic transformation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.2
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• pp.100-104
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• 2013

Zoysiagrass (Zoysia japonica Steud.) is an important forage and turfgrass that spreads by stolons and rhizomes. Zoysiagrass stolon can be used directly for Agrobacterium-mediated genetic transformation by exploiting the potential of direct shoot formation. However, surface sterilization of field-grown stolons is difficult and remains to be explored. We developed an effective surface sterilization and culture method using the stolon explant for infection with Agrobacterium tumefaciens. Among various treatments, sequential disinfection in 30% bleach for 15 min followed by 0.1% mercuric chloride for 25 min resulted in the highest number of clean stolons. The efficacy of mercuric chloride was increased under vacuum conditions by incubating at 800 mbar for 5 min. The inclusion of 2.5 mg/l amphotericin B further prevents fungal growth in in vitro cultures. This protocol would speed up the development of transgenic plants by utilizing field-grown stolon nodes.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.3
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• pp.191-198
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• 2005

In order to optimize tissue culture conditions for genetic transformation of orchardgrass, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of 3 cultivars, 'Frode'. 'Roughrider' and 'Frontier' as explant tissues. Callus induction medium containing 3mg/L 2,4-D or 3mg/L dicamba each with 0.1mg/L BA was optimal fer embryogenic callus formation from mature seed and had a strong effect on successive plant regeneration. The regeneration frequency from embryogenic callus among cultivars were descending order of Roughrider > Frode > Frontier. Supplementation of the regeneration media with 10mg/L $AgNO_3$ and 40mg/L cysteine enhanced frequency of plant regeneration. Efficient regeneration system established in this study will be useful fur molecular breeding of orchardgrass through genetic transformation.

• Korean Journal of Plant Resources
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• v.24 no.2
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• pp.121-129
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• 2011

As the molecular breeding was progressed, many plant transformation techniques were attained for improving transformation accuracy and used to produce useful transgenic plants. Day by day, new varieties were developed so new transformation techniques required for these newly developed varieties. Carnation (Dianthus caryophyllus L.) is a popular and economically important ornamental plant, all over the world. Keeping this in view, we selected 18 varieties of D. caryophyllus L. commonly available in the market and did optimization of culture conditions for more efficient tissue culture and to get higher number of plants via micro-propagation. Four varieties namely Yellowdotcom, Jakarta, Belmonte, Polartessino etc. were selected for organ culture studies from single cell line. The optimum growth was recorded in the MS media supplied with sucrose 3%, NAA 1.0 mg/L and TDZ 1.0 mg/L. except Belmonte, in which, BA 1.0 mg/L was found to be the best combination, in place of TDZ, rest ingredients were same. The most efficient coagulating agent used to obtain higher number of plant from callus was phytagel 0.3%. The most effective explant for higher shoot formation was stem in which 80.2% shoot formation was recorded. It also reduced culture periods by 6 days.

• Korean Journal of Plant Resources
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• v.20 no.4
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• pp.367-374
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• 2007

This study was carried out to establish the micropropagation system of Bupleurum latissimum Nakai that is a Korean native endangered species. Callus were induced from the leaf, petiole and floral bud and the percentage of callus formation was highest in the floral bud on the MS medium containing 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D. Especially, callus induced from floral bud was formed 77.8% and the percentage of shoot formation was 42.6% on the MS medium containing 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D plus 1.0 mg ${\cdot}$ $L^{-1}$ TDZ. For simultaneously callus formation and shoot regeneration, 1/2 MS medium was more effective than MS medium. The percentage callus formation, shoot regeneration and rooting were 46.3%, 13.0%, 13.0% in 1/2 MS medium, respectively. Soot regeneration from callus was good in 1/2 MS medium supplemented with 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D plus 1.0 mg ${\cdot}$ $L^{-1}$ BA where percentage of shoot regeneration was 74.1 %, and the number of shoot per explant was 2.4. The percentage of rooting was lowest (57.8%) in control while it was highest (97.8%) in 1.5 mg ${\cdot}$ $L^{-1}$ NAA. In acclimatization of regenerated plantlets, the percentage of survived plantlets was highest (86.1%), and plant height, root length and fresh weight were good in the soil for horticulture.

• Korean Journal of Medicinal Crop Science
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• v.5 no.4
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• pp.289-293
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• 1997

The effects of plant regulators on direct somatic embryogenesis in liquid culture of Rehmannia glutinosa were investigated and the proper explant for direct somatic embryo formation was studied. Direct somatic embryos were induced from leaf segments culture in the MS liquid medium containing 0.5 mg/l of both IAA and NAA, while IBA of 1.0 mg/l was required for the same effect. Many somatic embryos were directly formed at the concentration of 2.0 mg/l cytokinin such as BA, kinetin and zeatin, but somatic embryogenesis was relatively poor at above or below this level. Relatively more somatic embryos were induced in the combination of 1.0mg/l IAA and 2.0mg/l zeatin. Formation of somatic embryos begun after 6 weeks on stem segments, while 7 weeks both on petiole and leaf. However, overall production of somatic embryos after 8 weeks was higher in leaf segment than that of stem segment.

• Korean Journal of Medicinal Crop Science
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• v.10 no.1
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• pp.41-45
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• 2002

An efficient and reproducible procedure for the large scale propagation of Hovenia dulcis Thunb. is described. Shoot primodia emerging from the leaf surface was induced from MS medium supplemented with NAA. Stem cuttings were suitable explants for multiple shoot proliferation. They produced axillary shoots which branched repeatedly, yielding an average of 7 shoots per explants after 4 weeks in culture, when cultured on a woody plant medium (WPM) containing 0.1mg/l BA and 0.1mg/l NAA. Stem, leaf and root segments from axenic seedlings were used as explant source to induce somatic embryogenesis. A high frequency of somatic embryos were induced directly from leaf in MS medium with NAA, 2,4-D and in medium containing NAA, 2,4-D with BA. Somatic embryos were germinated in MS medium supplemented with 1mg/ l $GA_3$. Somatic embryos proliferated secondary somatic embryos rapidly after transfer to MS medium supplemented with 1mg/ l kinetin, 1mg/ l $GA_3$ and 2% dextrose.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.54-54
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• 2018

억새(Miscanthus spp.)는 우리나라 등 동아시아가 원산이며 바이오매스량이 많아 바이오에너지 원료작물로서 잠재성이 크기 때문에 2세대 바이오에탄올 생산 원료로 주목을 받고 있고, 축산깔개 및 토양개량제 등으로도 이용되고 있다. 미국과 유럽 등에서는 생태계 교란 방지를 위해 4배체 물억새(M. sacchariflorus)와 2배체 참억새(M. sinensis)의 종간 교잡 이질 3배체인 불임성 억새(M. x giganteous)을 주로 재배하고 있으나, 단일 유전형의 품종으로 병해충과 자연재해에 취약하여 다양한 억새 품종의 개발이 시급한 실정이다. 본 연구는 억새를 재료로 하여 반수체 및 배수체 확보를 통한 배수성 별 특성 평가와 함께 기내배양을 통해 탈분화 및 재분화 시스템을 구축하여 억새의 육종 효율을 높이기 위해서 실시하였다. 억새 종자로부터 캘러스의 유도는 MS배지와 N6배지에 1mg/L 2.4-D를 첨가하였을 때 비배발생 캘러스(nonembryogenic callus)가 유도되었고, N6배지에 3~5 mg/L 2,4-D를 첨가하였을 때 배발생 캘러스(embryogenic callus)가 발생하였다. MS배지보다는 N6배지에서 캘러스 유도율이 높았으며, 식물생장조절제로 2,4-D와 BA 조합처리 보다 2,4-D 단용 처리하였을 때 캘러스 유도율이 더 높았다. 억새 종에 따른 캘러스 유도율은 물억새가 25.2~49.3%, 참억새는 30.3~52.0%였고 거대억새는 62.6~81.1%로 나타났다. 억새 신초 및 줄기로부터의 캘러스 유도율은 물억새가 4.4~17.2%, 참억새는 1.8~7.7%, 거대억새는 15.3~19.9%로 나타나 종자에 비해 매우 낮았다. 미성숙화기로부터의 캘러스 유도는 억새 종에 따른 차이가 없었으며, 3mg/L 2,4-D 첨가 배지에서 캘러스 유도율이 비교적 낮았고(60~80%), 1mg/L와 5mg/L의 2,4-D가 첨가된 배지에서 캘러스 유도율이 높게(90~95%) 나타났다.