• Journal of The Korean Society of Grassland and Forage Science
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• v.26 no.2
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• pp.83-90
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• 2006

To develop transgenic birdsfoot trefoil (Lotus corniculatus L.) plants tolerant to environmental stress, Arabidopsis NDPK gene (AtNDPK) was introduced into birdsfoot trefoil plants using Agrobacterium-mediated transformation and expressed powerfully under the control of the E35S promoter. The expression vector, pEN-K was used for introduction of AtNDPK gene into birdsfoot trefoil plaits. The transformed calli were selected on kanamycin containing medium and then regenerated. The transformed birdsfoot trefoil plants were cultivated for 4 months on BOi2Y medium. Genomic DNA PCR and Southern blot analysis confirmed the incorporation of AtNDPK into the birdsfoot trefoil genome.

• Journal of The Korean Society of Grassland and Forage Science
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• v.37 no.3
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• pp.223-230
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• 2017

Arabidopsis nucleoside diphosphate kinase 2 (AtNDPK2) is an upstream signaling molecule that has been shown to induce stress tolerance in plants. In this study, the AtNDPK2 gene, under the control of a stress-inducible SWPA2 promoter, was introduced into the genome of tall fescue (Festuca arundinacea Schreb.) plants. The induction of the transgene expression mediated by methyl viologen (MV) and NaCl treatments were confirmed by RT-PCR and northern blot analysis, respectively. Under salt stress treatment, the transgenic tall fescue plants (SN) exhibited lower level of $H_2O_2$ and lipid peroxidation accumulations than the non-transgenic (NT) plants. The transgenic tall fescue plants also showed higher level of NDPK enzyme activity compared to NT plants. The SN plants were survived at 300 mM NaCl treatment, whereas the NT plants were severely affected. These results indicate that stress-inducible overexpression of AtNDPK2 might efficiently confer the salt stress tolerance in tall fescue plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.4
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• pp.267-274
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• 2005

To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, a thermptolerance gene, DgP23, was introduced into orchardgrass using Agrobacterium - mediated transformation method. PCR and Southern blot analyses using genomic DNA showed specific DNA band on agarose gel and hybridization signal on X- ray film in transgenic orchardgrass harboring the recombinant DgP23 gene, but not in the wild type and empty vector control plants. RT-PCR and Southern blot analyses using total RNA also showed specific DNA band and hybridization signal. Transgenic orchardgrass did not showed ny morphological aberration both in the green house and field cultivation. Thermotolerance of transgenic plants was not detected in laboratory test. but may detected in field test.

• Proceedings of the Korean Society of Grassland Science Conference
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• 2005.06a
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• pp.188-189
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• 2005

• Proceedings of the Korean Society of Grassland Science Conference
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• 2005.06a
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• pp.152-153
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• 2005

• Proceedings of the Korean Society of Grassland Science Conference
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• 2005.06a
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• pp.150-151
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• 2005

• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.3
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• pp.237-242
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• 2004

A system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of mature seed-derived embryogenic callus. Seed-derived calli were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase (HPT), neomycin phosphotransferase II (NPTII) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of $200\mu\textrm{M}$ acetosyringone (AS) in inoculation and co-culture media lead to a increase in stable transformation efficiency. Transformation efficiency was increased when embryogenic calli were co-cultured for 5 days on the co-culture medium. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agyobacterium in the presence of 0.1% Tween20 and $200\mu\textrm{M}$ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and Southern blot analysis of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.4
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• pp.281-286
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• 2005

To develop transgenic birdsfoot trefoil (Lotus corniculatus L.) plants tolerant to environmental stress, Arabidopsis NDPK gene (AtNDPK) was introduced into birdsfoot trefoil plants using Agrobacterium-mediated transformation and expressed powerfully under the control of the SWPA2 promoter. The expression vector, pCAMBIA2300 was used for introduction of AtNDPK gene into birdsfoot trefoil plants. The transformed calli were selected on kanamycin containing medium and then regenerated. The transformed birdsfoot trefoil plants were cultivated fur 4 months on BOi2Y medium. Genomic DNA PCR and Southern blot analysis confirmed the incorporation of AtNDPK into the birdsfoot trefoil genome.

• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.235-242
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• 2004

A system for the production of transgenic plants has been developed for Italian ryegrass(Lolium mult리orum Lam.) via Agrobacterium-mediated transformation of embryogenic callus. Mature seed-derived calli were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase(HPT), neomycin phosphotransferase II (NPTII) and intron-oontaining $\beta$g1ucuronidase( intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of 200${\mu}M$ acetosyringone(AS) in inoculation and co-cultivation media lead to a significant increase in stable transformation efficiency. Increasing Agrobacterium cell density up to 1.0 in $OD_{600}$ during infection increased transfonnation efficiency of embryogenic calli. The highest transfonnation efficiency was obtained when embryogenic calli were incoulated with Agrobacterium in the presence of 0.1% Tween20 and 200${\mu}M$ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and PCR analysis of transgenic plants demonstrated that transgenes were integrated into the genome of Italian ryegrass.

• Journal of The Korean Society of Grassland and Forage Science
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• v.26 no.2
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• pp.77-82
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• 2006

An efficient transformation system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of seed-derived callus. From the point of morphogenetic capacity, three types of callus were selected. High frequency of plant regeneration was obtained by selection of type II callus, and the plant regeneration frequency was 52.6% when embryogenic callus were cultured on the regeneration medium. Supplementation of the media with 10 mg/L $AgNO_3$ and 40 mg/L cysteine enhanced frequencies of plant regeneration up to 65.3%. The highest transformation efficiency was also obtained when type II callus were inoculated with Agrobacterium. Southern blot analysis of PCR products of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue. Efficient regeneration system and transformation established in this study will be useful for molecular breeding of tall fescue through genetic transformation.