• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11b
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• pp.30-30
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• 2002

꿩의비름 (Sedum erythrostichum)은 매우 우수한 지피식물이며 건조에 강한 대표적 식물로 바위정원 (rock garden)을 가꾸는데 있어서 중요한 수종으로 이용되며, 유럽등지에서는 지붕에 식재하기도 하며 최근에는 빌딩옥상녹화의 대표적 수종으로 식재되고 있다. 또한 한방에서는 경천이라 불리우기도 하는데 피부상처 치유 및 미백효과가 탁월하다고 알려져 있다. 본 연구에서는 Agrobacterium을 매개로한 꿩의비름의 형질전환 시스템을 개발하고 아울러 phosphinothricin-N-acetyltransferase (PAT) 유전자를 도입하여 제초제 저항 식물을 개발하고자 수행되었다. 꿩의비름 잎을 Agrobacterium에 담근후 0.5 mg/l NAA와 2 mg/1 BA가 첨가된 MS 배지에 3일간 공동배앙 하였다. 그 후 300 mg/1 cefotaxime이 첨가된 같은 배지에 옮겨 계대하면서 Agrobacterium을 제거하였다. 약 3주후에 잎 절편으로 부터 직접적으로 부정아가 형성되기 시작 하였는데 이 시기부터 잎 절편을 25 mg/1 kanamycin이 첨가된 선발배지에 옮겨 주었다. 이 결과 배양된 잎 절편 절편 중 3.75%에서 kanamycin에 저항하는 부정아를 얻을 수 있었다. 형질전환체는 X-gluc 반응, PCR, Southern, Nothern analysis를 통하여 확인하였다. 약 94%의 형질전환 식물체는 성공적으로 토양에 옮길 수 있었으며 약 3개월후에 꽃을 피웠다. 형질전환체는 제초제인 Basta ($^{(R)}$ phosphinothricine at 200 mg/1)를 살포하여 주었을 경우 생존함을 확인 하였다.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.147-152
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• 1998

AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.120-125
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• 1998

A plasmid pNPG contains a genomic DNA complementing npgA1 which is located on the left arm of linkage group I. It transformed Aspergillus nidulans at a high frequency. No abortive transformants were observed and the $Trp^+$ transformants were all $Npg^+$. The 10.4 kb Psti fragment of the genomic DNA was subcloned into pILJ16, which increased the transformation efficiency by more than 200-folds. The transformants were mitotically unstable and yielded $Arg^-$ conidia at the frequency of more than 80%. An additional gene cloned into the plasmid containing the fragment was always lost with $argB^+$ marker. These characteristics strongly indicate the possibility that the plasmids autonomously replicate. The full activity of enhanced transformation was retained on the 4.9 kb EcoRI-HaeIII fragment. The DNA segment was similar to AMA1 rather than ANS1 in function and designated AMA2.

• The Journal of the Convergence on Culture Technology
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• v.5 no.3
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• pp.327-332
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• 2019

Today's scientists try to remove heavy metals with many new technologies such as phytoremediation. One of the best cutting edge technologies is developing transgenic plants to remove certain heavy metal in soil. I constructed the transformation vector expressing T. goesingense Metal Transport Protein1 gene and TgMTP1: GFP genes. The transgenic plants were selected and confirmed the transformed genes into Arabidopsis thaliana genome. Expression was confirmed in several parts in Arabidopsis cells, tissues and organs. When TgMTP1 overexpressing Arabidopsis thaliana were subjected, transgenic plants showed higher heavy metal tolerance than non-transgenic. For further study I selected the transgenic plant lines with enhanced tolerance against four different heavy metals; Zn, Ni, Co, Cd. The accumulation of these metals in these plants was further analyzed. The TgMTP1 overexpressing Arabidopsis thaliana plant of selected lines are resistant against heavy metals. This plant is characterized by the expression of the MTP1 gene accumulating heavy metal in the vacuole and being simultaneously expressed on the plasma membrane. In conclusion, these plants may be used in plant purification applications, and as a plant with increased tolerance.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.313-317
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• 1997

Leaf segments of the potato (Solanum tuberosum L.) genotypes, ND860-2, Norchip, Russet Norkotah, Goldrush, and Norqueen Russet were transformed with the coat protein gene of potato virus Y (PVY). The white-skinned genotypes, ND860-2 and Norchip, were easily transformed and regenerated into shoots, whereas the three russet-skinned genotypes had low frequencies of regeneration. Transformed shoots were generally recovered in four to six weeks. Antibody to PVY coat protein detected a single band of 30 kD in western blots of transgenic plants. Transformed plants had a normal phenotype in the greenhouse and many showed a delayed buildup of PVY following inoculation. Several transgenic lines had negative ELISA readings 85 days after inoculation. Transgenic lines which did not show detectable levels of PVY antigen will be further tested for resistance to PVY.

• Journal of agriculture & life science
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• v.45 no.6
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• pp.175-181
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• 2011

Effective Agrobacterium-mediated transformation of Trichoderma harzianum could be achieved using the $Al_2O_3$ particles-abraded mycelia pellets. Transformation efficiency, as percents for the number of hygromcin-resistant mycelia pellets out of total pellets tested, was about 20 in average for $Al_2O_3$ experiment. No transformed mycelium was obtained from the intact mycelia pellets. After second round of antibiotics selection, DNA integration of hygromycin resistant gene and the expression of target gene could be confirmed by PCR and RT PCR, respectively. This is the first report of Agrobacterium-mediated T. harzianum transformation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.29 no.4
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• pp.299-306
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• 2009

Oxidative stress is the main limiting factor in crop productivity. To solve global environmental problems using the plant biotechnology, we have developed on the oxidative stress-tolerant transgenic tall fescue plants via Agrobacterium-mediated genetic transformation method. In order to develop transgenic tall fescue (Festuca arundinacea Schreb.) plants with enhanced tolerance to multiple environmental stresses, nucleotide diphosphate kinase gene under the control of CaMV35S promoter were introduced into genome of tall fescue plants. Proteomic analysis revealed that transgenic tall fescue not only accumulated NDP kinase 2 protein in their cells, but also induced several other antioxindative enzyme-related proteins. When leaf discs of transgenic plants were subjected to cold stress, they showed approximately 30% less damage than wild-type plants. In addition, transgenic tall fescue plants showed normal growth when transgenic plants were subjected to $4^{\circ}C$ for 3 days treatments. These results suggest that transgene is important in ROS scavenging by induction of antioxidative proteins, and could improve abiotic stress tolerance in transgenic tall fescue plants.

• Korean Journal of Medicinal Crop Science
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• v.9 no.2
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• pp.156-165
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• 2001

Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.269-272
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• 1997

Cotyledonary petioles of Brassica napus cocultivated with Agrobacterium vectors for 72 h were transferred to MS medium with 0.5 mg/L NAA, 2.0 mg/L BA, 30 mg/L kanamycin, 100 mg/L cefotaxime, 30 g/L sucrose, 3 mg/L $\textrm{AgNO}_{3}$ and 2 g/L Gelrite. The cotyledonary petioles with green shoots were selected at a frequency of 17.5% in a selection medium and then rooted. Southern blot analysis confirmed the rolC and NPT IIgenes were incorporated into the regenerated plants. The stable inheritance of rolC gene was confirmed in progeny test of transgenic plants.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.1-8
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• 2011

Recently the increasing of vinyl and green houses and development of reclaimed land including Saemangeum induced the need for breeding salt-tolerant crops which can survive and grow in high salinity soil. So we try to develop salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum.) lines by using anti-porter gene TANHX and HVNHX. Through marker selection and plant regeneration step, we could get 284 putative transgenic chrysanthemum lines. On selected putative transgenic plants, 40 candidates were used for genetic analysis and 30 lines could be made up of target size band on PCR, so about 75% of marker selected lines were decided as real transgenic lines. Selected 284 transgenic lines were also used for salt-tolerance test as a range of NaCl 0.2 ~ 1.2% (300 mM). As a result of salt-tolerance test, 15 selected transgenic lines could live and grow on the continuous supply of 0.8% (200 mM) NaCl solution and another 7 lines were could survive under 1.2% (300 mM) NaCl solution. This salt-tolerant transgenic lines under salt stress also lead a cell alternation especially a guard cell. A stressed guard cell be swelled and grow larger in proportion to NaCl concentration. TTC test for cell viability on transgenic chrysanthemum lines pointed out that more strong salt-tolerant lines can be live more than another under same salt stress. The numerical value of strong salt-tolerant 7 transgenic lines were 0.206 ~ 0.331 under 1.2% NaCl stress, and then it's value is more larger than middle salinity lines' 0.114 ~ 0.193 and non-transgenic's 0.046. And the proline contents as indicated stress compound also pointed out that HVNHX introduced salt-tolerant transgenic lines were less stressed than other under same salt stress. The contents of strong salt-tolerant transgenic lines were 2.255 ~ 2.638 mg/kg and it is much higher than that of middle salinity lines' 1.496 ~ 2.125.