• Journal of Plant Biotechnology
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• v.37 no.3
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• pp.337-342
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• 2010

To establish the optimum conditions of in vitro plant regeneration, the leaf, rhizome, and root explants of Belamcanda chinensis were cultured on the MS medium supplemented with different concentration of 2,4-D and BA. The callus induction was more effective in the root explants than the leaf and rhizome explants, and was the best on MS medium containing 3.0 mg/L 2,4-D or 1.0 mg/L 2,4-D and 3.0 mg/L BA. The highest numbers of shoots were regenerated when callus were cultured on MS medium containing 3.0 mg/L 2,4-D for 4 weeks. However, the multiple shoots were induced on MS medium supplemented with the combination of 2,4-D and BA. The normal root formation from shoot was effective on the MS medium lacking any plant growth regulators. For acclimatization, the regenerated plantlets were cultured on MS medium without sucrose and plant growth regulators for 2 weeks, and then transferred to the pot.

• Journal of Plant Biotechnology
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• v.31 no.1
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• pp.73-78
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• 2004

This study was carried out to develop an in vitro chromosome doubling systems for the breeding of tetraploid lily cultivar. Different concentration (ppm) and treatment time (hour) of colchicine, oryzalin and caffeine were compared for the efficiency of bulb regeneration and tetraploidization. The occurrence of tetraploid plants by colchicine was the highest in the concentration of 1,000 ppm for three hours. In oryzalin treatment, the best combination was at 30 ppm for three hours. However, the effects of oryzalin treatments were similar between 10 ppm and 30 ppm concentrations. The survival rate was dramatically reduced in a high concentration of caffeine although some treatments had a higher tetraploid induction. As a consequence, reliable results for the tetraploid production were obtained in the treatments of 1,000 ppm colchicine, 3,000 ppm oryzalin both for three hours, and 9,000 ppm caffeine for nine hours.

• Korean Journal of Plant Resources
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• v.26 no.2
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• pp.284-288
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• 2013

This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.207-217
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• 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 Life Science
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• v.14 no.4
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• pp.644-649
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• 2004

During plant development, active gibberellins (GAs) control many aspects of plant growth and development including seed germination, stem elongation, flower induction, anther development and seed growth. To understand the biosynthesis and functional role of active GAs in high plants, this study investigated GA 3$\beta$-hydroxylase gene en-coding $GA_1$ and$GA_4$ catalizing last step in GA biosynthetic pathway. The antisense GA 3$\beta$-hydroxylase gene was inserted into expression vector, pIG121-Hm. Calli derived from mature seeds of rice (Oryza satiiva L. cv. Donjinbyeo) were co-cultivated with Agrohacterium tumefaciens EHA101 earring a pIG121-Hm containing hygromycin resistance ($Hyg^r$) and antisense GA 3$\beta$-hydroxylase gene. Seventeen transgenic plants obtained inhibiting GA 3$\beta$-hydroxylase. Transgenic plants had shorter plant height more than that of the Dongjinbyeo. Stable integration of antisense GA 3$\beta$-hydroxylase gene was confirmed by polymerase chain reaction of genomic DNA isolated from the leaf organs of the $T_o$ generation.

• Horticultural Science & Technology
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• v.17 no.6
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• pp.770-772
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• 1999

This study was conducted to provide a basic system to develop a molecular marker for plant cultivar protection using a recombinant DNA technology. Using Nicotiana tabacum L. plants, the potentiality in the utilization of the developed marker was examined. After homology test with several plant genomes, mouse adenosine deaminase (ADA) gene was selected as DNA source of a molecular marker for cultivar protection. As a result of the digestion of ADA gene with BamHI and Pst I, six DNA fragments were obtained, and 513 bp DNA fragment among them was selected as a possible DNA marker for cultivar protection. Selected 513 bp DNA fragment was efficiently inserted into pBI101 plasmid vector for plant transformation by using phagemid vector pBluescript II SK (+/-) as an intermediate vector. The recombinant pBI101, carrying 513 bp DNA fragment, possible markers for cultivar protection, was transformed into A. tumefaciens LBA4404. Nicotiana tabacum was transformed with A. tumefaciens LBA4404 having the recombinant pBI101 and was confirmed the transfer of 513 bp DNA fragment, a possible molecular marker for cultivar protection.

• Korean Journal of Plant Tissue Culture
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• v.25 no.2
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• pp.119-123
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• 1998

In order to investigate the possibility of in vitro mass propagation via somatic embryogenesis from Sicyos angulatus L., effects of plant growth regulators and carbon sources on callus induction and somatic embryogenesis were evaluated. Optimal combinations of plant growth regulator for callus induction from cotyledon and inflorescence explants were 2,4-D 2.0 mg/L + BA 0.1 mg/L and 2,4-D 1.0 mg/L + BA 0.1 mg/L in MS basal medium supplemented with sucrose 30 g/L,, respectively. Somatic embryogenesis was observed from cultured inflorescence explants, but it could not be achieved from leaf or cotyledon explants. The most effective plant growth regulators for somatic embryogenesis from callus was NAA 1.0 mg/L + kinetin 10 mg/L in the half strength of MS basal medium supplemented with 20 g/L sucrose.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.21-21
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• 2018

본 연구는 북방계 식물에 속하는 두메우드풀[Woodsia ilvensis (L.) R.Br.]의 전엽체 증식 및 포자체 형성을 위한 적정방법을 구명하고자 수행되었다. 식물재료는 경상북도 의성군 일대에서 채취한 후 청주의 일반하우스에 이식하여 재배하였다. 포자 성숙기인 8월에 채종하여 발아시켰으며, 포자로부터 획득한 전엽체를 8주간 계대하면서 실험재료로 사용하였다. 전엽체 300mg을 메스로 다진 후 배지농도 및 종류(Knop, 1/4, 1/2, 1 및 2MS)를 달리하여 8주간 배양하였다. 배양실의 온도는 $25{\pm}1.0^{\circ}C$, 광도는 $30{\pm}1.0{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$(16/8h)로 조절되었다. 연구의 결과, 배지구성물질의 농도가 높았던 2MS배지에서 전엽체의 생체중이 12.4g으로 41배 증가하여 가장 높은 증식율을 나타냈다. 또한 배지구성물질의 농도가 적을수록 생체중의 증가폭도 함께 감소하는 경향을 보였다. 포자체 형성에 적합한 토양조건을 확인하고자, 원예상토, 피트모스, 펄라이트 및 마사토의 비율을 달리한 5종류의 혼합토양을 조성하여 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하였다. 다음으로 전엽체 1g을 핸드블랜더로 10초간 분쇄 후 충진된 사각분 위에 고르게 분주하여 14주간 재배하였다. 재배환경은 온도 $25{\pm}1.0^{\circ}C$, 광도 $43{\pm}2.0{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$(16/8h) 및 습도 $72{\pm}2.0%$로 조절되었다. 지베렐린 수용액이 포자체 형성에 미치는 영향을 조사하고자, 선행연구에서 선발된 토양조건을 기준으로 침지농도(0, 50, 100, $200mg{\cdot}L^{-1}$)를 조절하여 1시간 침지 후 상기와 동일한 방법으로 재배하였다. 연구의 결과, 원예상토와 펄라이트 또는 마사토를 2:1(v:v)로 혼용한 토양에서 사각분 당 109.5, 128.8개의 포자체가 형성되었다. 한편, 피트모스가 첨가된 처리는 포자체의 형성이 억제되는 경향을 나타냈다. 지베렐린 50과 $100mg{\cdot}L^{-1}$ 수용액에 침지한 처리에서 무처리에 비해 많은 사각분 당 각 177.0, 181.7개의 포자체가 형성되었으며, 생육도 양호하였다.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.82-88
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• 2017

Transgenic lily plants have been obtained after particle bombardment, using PDS-1000/He system and scale explants of lilies, followed by PPT (D-L-phosphinothricin) selection. In this study, scales of the lily plants cv. 'red flame' were bombarded with a plasmid containing the bar gene as a selectable marker, and the AtSIZ gene as a gene of interest, showing salt tolerance and drought tolerance respectively, and both being driven by the CaMV 35S promoter. For optimization of a protocol, factors which optimized and showed a high transformation efficiency under following conditions, were considered: a bombardment pressure of 1100 psi, a target distance of 6 cm and $1.0{\mu}m$ of gold particle, and 24-h pre-culture and post-culture on MS medium containing 0.2 M sorbitol and 0.2 M mannitol as osmoticum agents. After bombardment, all the bombarded scales of lily were transferred to MS medium without selective agents, for a week. Subsequently, these bombarded scales were transferred to a selection MS medium containing 10 mg/l PPT, and incubated for a month for further selection, after which they were cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. After transferring into hormone-free MS medium, the PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets. PCR analysis revealed that the surviving putatively transformed plantlets indicated the presence of both the bar gene and the AtSIZ gene. In conclusion, when 100 scales of lily cv. Red flame are bombarded, this study produced approximately 17-18 transgenic plantlets with an optimized bombardment protocol. The protocol described here can contribute to the breeding program of lilies.

• Korean Journal of Plant Resources
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• v.18 no.1
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• pp.15-21
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• 2005

Korean ginseng(Panax ginseng C.A. Meyer) is very difficult to obtain stable production of qualified ginseng roots because of variable stresses in soil environments. In transformation of ginseng with betain aldehyde dehydrogenase gene, compounds synthesized for controlling osmotic pressure such as proline, glycine, betaine, polyols and sugar were accumulated in cell for salt resistance in transgenic plants. 2 Agrobactgerium conjugants were acquired with bet A and bet B genes for solt resistant plants. A. tumefaciens MP90/pBetA and A. tumefaciens MP90/pBetB were recombined for increasing the tolerance to salt stress. To confirm the transformation of the binary vector, tobacco plant was transformed, and the transformant can grow on media containing high concentration of kanamycin. To identify NPT 11, BetA and BetB genes of the transformants, the band on the agarose was confirmed by PCR and RT-PCR techniques. The transformants of ginseng with bet A and bet B genes were acquired on the phytohormone free basic MS media containing only antibiotics and 1M mannitol used for selection of transgenic plant, but the transfomation efficiency for BetA and BetB was very low.