• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.69-74
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• 2008

This study aims to establish the efficient soybean transformation system and develop soybean [Glycine max (L.) Merill] transformants using cotyledonary node explants. The cotyledonary node of soybean were co-cultivated with Agrobacterium tumefaciens strains (KYRT1, EHA105). These strains contain the binary vector pCAMBIA3301 which carries a herbicide-resistant far gene. Korean cultivars (Danbaekkong, Eunhakong) and foreign cultivars (Jack, Peking) were the most efficient in regenerating cotyledonary node. Therefore, they were chosen for the transformation. Results showed that the T-DNA transfer reached up to 60% and transformation efficiency reached up to 3% in the cotyledonary node explants from Jack cultivar, co-cultivated with EHA105 strain. Histochemical GUS evaluation showed that 12 individual lines, transformed with the 현 gene, have positive response. The transformed soybeans have been confirmed in the $T_0$ generation through phenotypic assay using herbicide $Basta^{(R)}$ and Southern blot analysis.

• Korean Journal of Medicinal Crop Science
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• v.13 no.1
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• pp.1-5
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• 2005

While the rusty-colored root is common in ginsengs culture and, often results in a severe economic loss, the major factors have not been found. This study was focused on the determination of a potential relationship between rusty root and endophytic bacteria. The number of endophytes was $9.6\;{\times}\;10^1{\sim}1.5\;{\times}\;10^2\;cfu/g$ fw in normal ginseng roots compared to $3.7\;{\times}\;10^6{\sim}5.1\;{\times}\;10^7\;cfu/g$ fw in rusty ones. Of 31 isolates from rusty ginseng roots, twenty-four isolates repeatedly induced severe to moderate rust on root while seven isolates induced slight rust. The bacteria responsible for rusty ginseng roots were mainly Gram negative aerobic. Rust inducing bacteria were identified as Agrobacterium tumefaciens, A. rhizogenes, Burkholderia phenazinium, Ensifer adharens, Lysobacter gummosus, Microbacterium luteolum, M. oxydans, Pseudomonas marginalis, P. veronii, Pseudomonas sp., Rhizobium leguminosarum, R. tropica, Rhodococcus erythropolis, Rh. globerulus, Variovorax paradoxus on the basis of bacteriological characters and 16S rDNA sequences analysis. The results in this study strongly suggested that the rusty ginseng roots were produced by infection and growth of endophytic bacteria.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.93-98
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• 2002

A VP6 fragments was subcloned with BamHI in the binary pMBP-1 vector under Califlower Mosaic Virus (CaMV) 355 promoter and neomycin phosphotransferase II (npt II) gene. The recombinant binary vector was mobilized into Agrobacterium-tumefaciens LBA4404 by the freeze-thaw method and potato (Solanum tubensum L. cv Desiree) was transformed by modified leaf-disc cocultivation. Shoots were induced on MS medium with 0.01 mg/L NAA, 0.1 mg/L GA$_3$, 2.0 mg/L Zeatin, 100.0 mg/L kanamycin, 500.0 mg/L carbenicillin. In order to identify the copy number of VP6 into potato plant, total genomic DNA was isolated from transgenic potato and analysed by Southern blotting. Genomic DNA and total mRNA analysis demonstrated the incorporation of the foreign gene into the potato genome, as well as their transcription.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.102-109
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• 2010

Fructan has been found to accumulate in various tissues during periods when light levels increased carbon fixation where low temperatures reduced growth rates while photosynthesis continued. In this study, we have cloned 1-sucrose:sucrose fructosyl transferase(1-sst) and 1-fructan: fructan fructosyl transferase (1-fft, a key enzyme for the synthesis of fuctan) from Jerusalem Artichoke (Helianthus tuberosus L.). The recombinant vector with 1-sst and 1-fft has been constructed under the control of 35S promoter of KJGV-B2 vector and transgenic plants obtained by Agrobacterium tumefaciens LBA4404. PCR analysis carried out on the putative transgenic plants for amplification of the coding region of specific gene (1-sst, 1-fft), and HPT genes. Transgenic lines carrying of 1-sst and 1-fft were confirmed for integration into the rice genome using Southern blot hybridization and RT-PCR. The transgenic plants in $T_2$ generation were selected and expression pattern analysis revealed that 1-sst and 1-fft were stable. This analysis confirmed the presence of low-molecular-weight fructan in the seedling of the transgenic rices. Therefore, cold tolerance and carbohydrate metabolism will be possible to develop resistant plants using the transgenic rice.

• Korean Journal of Microbiology
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• v.48 no.3
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• pp.207-211
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• 2012

Autoinduction is mediated by signaling molecules known as autoinducers (AIs) that are produced, released and detected by bacterium itself. We recently reported that Corynebacterium glutamicum possesses an autoinduction system which secretes autoinducers during the stationary-phase of growth, triggering the expression of acyltransferase gene. However, it is still not clear what may act as autoinducers for the autoinduction in C. glutamicum. In this study, we compared the inducing effects of cell-free culture fluids obtained from a number of microbes including Agrobacterium tumefaciens, Vibrio harveyi, and Escherichia coli. Fluids from A. tumefaciens did not increase the expression of acyltransferase, whereas fluids from V. harveyi BB120 ($AI-1^+$, $AI-2^+$) did. Interestingly, the expression was increased by the fluids obtained from the early exponential-phase culture of BB120. Furthermore, this induction was not observed by the fluids from autoinducer mutants of V. harveyi MM77 ($AI-1^-$, $AI-2^-$) and BB152 ($AI-1^-$, $AI-2^+$). Unlike the effect shown by BB152, fluids from E. coli ($AI-1^-$, $AI-2^+$) still induced the acyltransferase expression. Taken together, these results suggest that C. glutamicum autoinducers seem to be unidentified molecules which do not belong to AI-1 or AI-2.

• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.482-487
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• 2009

The apple rootstock M.26 (Malus pumila) is very popular apple rootstock with semi-dwarf habit and the trees on M.26 produce high quality fruit at a young age. Since it is prone to poor prop ability to soil, however, young trees require staking in windy locations. The rolC gene was introduced into M.26 by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain its transformants with dwarfism and enhanced rooting ability. One regenerated transgenic line was confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of rolC gene. The characteristics of transgenic line in vitro were not significantly different from non-transgenic line except for the active root formation and lateral root number. The rolC transgenic line showed reduced stem length and increased root number in vitro. Rooting ability was examined in the isolated greenhouse after mound layering. Compared to non-transgenic M.26, rolC transgenic line showed significantly higher rooting ability. The transgenic line did not show any other observable variation in shoot phenotype compared with non-transgenic line excepting increased branching

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.445-451
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• 2021

In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

• Korean Journal of Microbiology
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• v.4 no.2
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• pp.5-10
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• 1966

The relationships between tumor score and peroxidase activities of tomato stems infected with Agrobacterium tumefaciens strain A6Kl, B6, T372 11BNV6, 11BV7 and wounded stem as a control were examined in relation to crown gall tumor development on purpose to study the lignification of tumor tissue which is affected to the development of crown gall tumor. As the previous paper has been mentioned the fact that the induction of tumor tissues were inhibited or limited in the lignified stem of host plant. It was presumed that the activities of peroxidase related to the development of lignification were decreased during the period of tumor development. But the experimental result in this experiment shows that the peroxidase activities of crown gall tumor-tissues infected with the A. tumefaciens strains which are already known as virulent are increasing during four weeks, however, in the strain 11BNV6 and wound the peroxidase activities are decreasing on the second week after the inoculation of the bacteria strains. These results could be explained on the basis of that possible regulatory agents of lignification which were accumulated in tumor tissues, IAA, ascorbic acid, glutathion(GSH) and caffeic acid esters, were postulated to act as antioxidants which has been suggested by Stafford. Total nitrogen contents in relation to crown gall tumor development were determined for the detection of protein synthesis related to the enzyme activities which are increasing in the time of plant growth. Generally six groups are contained the largest amount of nitrogen on the second week after the inoculation of the bacterium. Comparing to the tumor score, it is presumed that the all of enzyme activities including peroxidase in tumor tissues are increasing from the second week through the third week after the inoculation of bacterium and the protein synthesis is stimulated under the most appropriated temperature during the above periods.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.435-439
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• 2000

Explants of lettuce (Lactuca sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring ${\gamma}$-tocopherol methyltransferase (${\gamma}$-TMT) gene from Arabidopsis thaliana. These explants were transferred to MS medium supplemented with 50 mg/L kanamycin, 500 mg/L carbenicillin, 0.1 mg/L NAA and 0.5 mg/L BA. After 4 weeks, kanamycin resistant shoots were obtained from the explants on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 50 mg/L kanamycin and 250 mg/L carbenicillin. Stable incorporation of the Arabidopsis ${\gamma}$-TMT cDNA into lettuce genomic DNA was confirmed by PCR and Southern analysis. HPLC analysis showed that $\alpha$- to ${\gamma}$-tocopherol ratio increased over four fold in a transgenic lettuce line indicating successful expression of the transgenic Arabidopsis ${\gamma}$-TMT in lettuce.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.441-445
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• 2000

In order to improve phosphate use efficiency of rice using phosphate transporter (PT), transgenic rice plants containing a tobacco PT gene were developed. Calli from Dongjinbyeo (Oryza sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring PT gene. Multiplied calli were transferred to MS medium supplemented with 50 mg/L hygromycin, 500 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA. After 2 weeks, hygromycin resistant shoots were obtained from the calli on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 250 mg/L cabenicillin. Plant regeneration rate from the calli was about 52%. Stable incorporation of the tobacco PT gene into rice genomic DNA was confirmed by PCR and Southern blot analysis.