• Plant Resources
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• v.6 no.2
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• pp.108-113
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• 2003

This study was carried out to obtain basic information for possibility of oral vaccine in carrot using Agrobacteruim -mediated transformation system. The epitope region of porcine epidemic diarrhea virus (PEDV) spike gene which is classified as a member of the Coronaviridae and causes an acute enteritis in pigs was successfully expressed in carrot (Daucus carota) using the Agrobacterium-mediated transformation system. Hypocotyl segments of in vitro germinated plantlets were infected with Agrobacteriun tumefaciens LBA 4404 harboring PEDV spike gene. Embryogenic callus (EC) was induced on MS selection medium with 1 mg/L 2,4-D, 50 mg/L kanamycin and 300 mg/L cefotaxime after 45 days of culture. Subcultured ECs on MS selection medium without 2,4-D were converted to somatic embryos (SE) of various stage; globular, heart and torpedo stage. Putative transgenic embryos were selected on MS medium with 50 mg/L kanamycin and 300 mg/L cefotaxime. Regenerated plantlets from transformed SE were induced on MS medium containing 50 mg/L kanamycin after 30 days of culture. Genomic PCR confirmed the integration of PEDV spike gene into nuclear genome of carrot and northern blot analysis demonstrated the expression of PEDV spike gene in transgenic carrot.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.341-346
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• 2003

In an effort to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), an efficient and high-frequency plant regeneration system from seed-derived calli was established. Embryogenic calli induced on MS medium containing 3mg/L 2,4-D and 0.1mg/L BA had significantly improved regeneration ability. Plant regeneration rate was 92％ when embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among three kinds of medium, MS and N6 medium were optimal for embryogenic callus induction and plant regeneration, respectively. Ho difference in callus induction frequency was observed among four cultivars of orchardgrass, however, "Roughrider" cultivar showed higher regenerability with the frequency of 61％. Addition of maltose to the regeneration medium as a carbon source dramatically increased regeneration frequency up to 69％. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of orchardgrass through genetic transformation.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.9
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• pp.911-916
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• 2005

In this study, variations of water quality, sediment size and contaminant substances are analyzed at upstream and downstream in sewer systems in order to evaluate the characteristics of wastewater transformation by physico-chemical and biological reactions. The differences of DOC concentration between up and down stream showed the range of $-5.8{\sim}18.6$ from the result of continuous measurement at up and down stream. About 8.4% of DOC concentration was reduced and reduction rate was 2.3 mg/L/km. SS reduction rate was measured by 5.5 mg/L/km, 0.22 mg/L/min from upstream to downstream, which was twice than DOC reduction rate. When pollution load reduction was evaluated considering infiltration/inflow effect, DOC load was eliminated from 1,230 ka/d to 1,167 kg/d by physi-chemical and biological reaction in a sewer and 7.8% of the SS in upstream station was reduced under dry weather condition. The results showed that the characteristics of transformation process of wastewater in sewer should be considered for design and operation of wastewater treatment plant.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.77-85
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• 2004

Development of efficient plant regeneration and genetic transformation protocols (using the Particle Inflow micro-projectile Gun and the shoot-tips as target tissue) of Sorghum bicolor (L.) Moench in terms of expression of the reporter gene, $\beta$-glucuronidase(uidA) is reported here. Two Indian cultivars of sorghum were used in the study, viz. M-35-1 and CSV-15. Plant regeneration was achieved from one-week-old seedling shoot-tip explants via multiple-shoot-clumps and also somatic embryos. The multiple-shoot-clumps were produced on MS medium containing BA (0.5, 1.0 or 2.0 mg/$L^{-1}$), with biweekly subculture. Somatic embryos were directly produced on the enlarged dome shaped expansive structures that developed from shoot-tip explants (without any callus formation) when cultured on MS medium supplemented both with BA (0.5, 1.0 or 2.0 mg/$L^{-1}$) and 2,4-D (0.5 mg/$L^{-1}$). Whereas each multiple-shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing 1.0mg/$L^{-1}$ IBA and successfully transplanted to the glasshouse and grown to maturity with a survival rate of 92%. The plant regeneration efficiency of both the genotypes were similar. After the micro-projectile bombardment, expression of uidA gene was determined by scoring blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The optimal conditions comprising a helium pressure of 2200 K Pa, the target distance of 11 cm with helium inlet fully opened and the use of osmoticum have been defined to aid our future strategies of genetic engineering in sorghum with genes for tolerance to biotic and abiotic stresses.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.187-192
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• 1994

Wheat (Triticum aestium L.cv Cho-Kwang) explants were transformed by electrporation. Excised root segments were elechoporated with plasmid DNA of pBI121 and transferred to medium containing 300 mg/L kanamycin. Transformed calli formed within 5-7 days of culture and were selected from electroporated tissue on medium containing kanamycin after 4 weeks. The highest transformation frequency was obtained after electroporation with a pulse of 200 V/800 uF and calli formed at frequencies up to 2.5%. GUS ($\beta$-glucuronidase) assay and dot blot analysis showed that the foreign gene was capable of expressing in root explants subjected to electroporation and calli derived from the explants..

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.52-52
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• 2018

Development of herbicide resistant transgenic legume plants through Agrobacterium-mediated transformation has been worked in many previous studied. Plant regeneration after infection is the important step to obtain successful transgenic plants. Many attempts try to find the optimum media condition for plant regeneration after infection. However, the transformation efficiency of legume plants is still low. In this study, regeneration of some Korean legume species including two soybean cultivars (Dawon and Pungsan) and pea have been done with organogenesis which is used various kind of explants such as cotyledonary-nodes in soybean and bud-containing tissue in pea. We developed the optimum media condition for plant regeneration regulators under Agrobacterium-mediated transformation using different kind and various concentration of plant growth. As the results, B5 medium containing 2 mg/L of 6-benzylaminopurine was selected in this study for the optimum plant regeneration media. The segments were inoculated with Agrobacterium suspension harbored an IG2 vector containing bar gene which confers resistance to phosphinotricin (PPT) in 3, 5 and 7 days. The transformation efficiency was achieved in Dawon 3.03 % and pea 1.46 % with co-cultivation period of 7 days which is showed a high number of GUS positive expression period.

• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.66-75
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• 2016

Agrobacterium-mediated gene transfer has recently been developed to improve rice transformation. In this study, 3 different transformation methods were tested including soaking, co-cultivation, and vacuum infiltration. Agrobacterium tumefaciens GV3101 harboring the binary vector pGreen:: LeGSNOR was used in this experiment. This study aimed to identify the most appropriate method for transferring LeGSNOR into rice. Vacuum infiltration of the embryonic calli for 5 min in Ilpum resulted in high transformation efficiency based on confirmation by PCR, RT-PCR, and qRT-PCR analyses. In conclusion, we described the development of an efficient transformation protocol for the stable integration of foreign genes into rice; furthermore, the study results confirmed that PCR is suitable for efficient detection of the integrated gene. The vacuum infiltration system is a potentially useful tool for future studies focusing on transferring important genes into rice seed calli, and may help reduce time and effort.

• KSBB Journal
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• v.6 no.3
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• pp.223-230
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• 1991

Using the shuttle vector pECCGl between Escherichia coli and Corynebacterium glutamicum and C. glutamicum strain JS231 grown in the medium supplemented with penicillin-G, which inhibits the formation of cross-links in the peptidoglycan of bacterial cell wall, various parameters involved in electroporation system including resistance, electric field strength, capacitance, DNA concentration, and cell density were investigated independently and optimized for the high efficiency transformation of coryneform bacteria. Using cells grown with 0.3U/ml of penicillin-G and harvested at A600 of 0.7-0.8, transformation efficiencies of 107-l08 transformants/$\mu\textrm{g}$ of DNA with Corynebcctertum glutamicum strain JS231 and wild type ATCC13032 were achieved under conditions of 12.5kV/cm of electric field strength, 400 ohms of resistance, $25\mu$F of capacitance, 3$\times$108 cells per transformation(1.2$\times$1010 cells/ml) and 100ng of plasmid DNA per transformation.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.388-393
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• 2017

We report an Agrobacterium-mediated transformation procedure optimized for 'Kyoho' that is a major table grapevine cultivar in Korea, and its transgenic plants with antifreeze protein gene of carrot (DcAFP). The full length of DcAFP coding region in accordance with the previous report was isolated from young leaves of carrot and recombined into a plant transformation vector. Ethylene inhibitors such as silver nitrate and aminoethoxyvinylglycin (AVG) supplemented in a co-cultivation medium distinctly increased frequency of shoot regeneration when explants were sub-cultured in a selection medium: particularly ten-fold higher in treatment with 0.1 mg/L AVG than one without ethylene inhibitor. Among various antibiotics and their concentrations, the combination of 150 mg/L cefotaxime plus 150 mg/L $Clavamox^{TM}$ was selected for elimination of Agrobacterium cells in addition to minimization of adverse effect on shoot regeneration, while 50 mg/L kanamycin monosulfate effectively suppressed regeneration of non-transgenic shoots. Applying the elucidated culture condition, we finally obtained a total of 5 transgenic 'Kyoho' plantlets with DcAFP, of which integration with the grapevine genome and transcription was confirmed by nucleic acid analyses.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.947-954
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• 2015

To select suitable spray chrysanthemum cultivars for Agrobacterium-mediated transformation, thirty-nine (39) spray cultivars bred in the National Institutes of Korea and a standard cultivar Jinba from Japan were collected and tested for regeneration rate and Agrobacterium infection assays. MS medium with $0.5mg{\cdot}L^{-1}$ IAA and $1.0mg{\cdot}L^{-1}$ BAP was used for shoot regeneration from leaf disks and internodes. The shoot regeneration rate in leaf disks was the highest in cultivar BRM, followed by cultivars VS, WW and YTM. The cultivar JB (Jinba) used as a transformation material in previous reports ranked similarly to cultivars PK and SPP. In shoot regeneration from internodes, the shoot regeneration rate was the highest for cultivar PA, followed by cultivar WW. The infection rate of leaves and internodes of 40 chrysanthemum cultivars with agrobacterium was investigated. Cultivars WPP, YNW, VS, PP, WW, FA, PA and YMN showed the highest infection levels in leaves, whereas cultivars WPP, PA, PK and YNW had the highest infection levels in internodes. Considering all of these results, cultivars VS and WW were the most appropriate for gene transformation of chrysanthemum using leaves, while cultivar PA was for internodes.