• 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.

• Proceedings of the Korean Society of Grassland Science Conference
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• 2003.06a
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• pp.172.1-172
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• 2003

• 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.

• Proceedings of the Korean Society of Grassland Science Conference
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• 2002.09a
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• pp.25.1-25
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• 2002

• Journal of The Korean Society of Grassland and Forage Science
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• v.28 no.3
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• pp.171-176
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• 2008

Efficient plant regeneration system of birdsfoot trefoil (Lotus corniculatus L.) was development. The factors affecting the somatic embryo formation, its proliferation and regeneration capacity of leaf and stem explants of Empire cultivar was investigated. The highest number of somatic embryos was obtained on B5 medium supplemented with 1 mg/L NAA and 1 mg/L BA. Depending on different explants, highest frequency of embryogenic callus and regeneration were observed in Empire with leaf explants. The response from stem explants was slower and callus induction was less than that from leaf explants. Regenerated shoots formed complete plantlets in on 1/2 MS medium supplemented with 1 mg/L IBA. Regenerated plants were morphologically uniform with normal shape and growth pattern.

• Journal of The Korean Society of Grassland and Forage Science
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• v.27 no.4
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• pp.229-234
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• 2007

To select the most proper soil for root initiation from stem cuts of Birdsfoot trefoil (Lotus corniculatus L.), eight-week-old stem cuts were cultured on three types of soil [commercial bed soil, decomposed granite (DCG), and river sand] for one month. The results showed that the root initiation ratios on DCG (77.8%) and river sand (70.0%) were relatively high, but the ratio on commercial bed soil (41.1%) was very low. To examine the effect of rare earth (RE) and Indole-3-Butyric Acid (IBA) on root initiation from stem cuts of Birdsfoot Trefoil, stem cuts were cultured on two types of soil (DCG and river sand) with treatment of RE and IBA for one month. The root initiation ratios turned out to be 90.0% (DCG with 60 ppm of RE), 80.0% (river sand with 20 ppm of RE), 96.7% (DCG with 40 ppm of IBA), and 96.7% (river sand with 40 ppm of IBA). These results suggested that the most efficient way for root initiation of Birdsfoot trefoil was to culture the stem cuts on river sand or DCG over 30 days with IBA treatment (40 ppm).

• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.4
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• pp.303-308
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• 1999

The conditions for callus formation and plant regeneration were confirmed in birdsfoot trefoil (Lotus corniculatus L.). Among SH (Schenk and Hildebrandt), MS (Murashige and Skoog) and N6 medium (Chu), SH medium was highest degree of efficiencies respectively in callus formation and plant regeneration. In this study, we determined volume of hormones and other compounds appended in media. For callus formation, only $3mg/{\ell}$ of 2,4-D (2,4-dichlorophenoxy acetic acid) was appended in their media. For plant regeneration, we used BOi2Y medium (Bingham et al.). We obtained birdsfoot trefoil plants from callus by regeneration, about sixty days later transfer calli to regeneration media.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.1
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• pp.35-38
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• 2001

When root initiation ratio of cut Birdsfoot trefoil (Lotus corniculatus L.) stems was examined in several medium conditions containing different IBA concentration, the higher IBA contration of medium was elucidated superior then lower IBA concentration. Highest root initiation ratio was confirmed at 2.5 mg/$\ell$ of IBA and the ratio was 90~95%. When the stems from regenerated shoots from callus were treated at 8 kinds of medium for 12 days, the root iniation result was 9 (45%) at 1/2 SH-0 medium, 10 (50%) at SH-0 medium, 10 (50%) at SH-0.5IBA medium, 10 (50%) at SH-1.0IBA medium, 13 (65%) at SH-1.5IBA medium, 14 (70%) at SH-2.0IBA medium, 19 (95%) at SH-2.5IBA medium and 14 (70%) at SH-3.0IBA medium.

• Journal of The Korean Society of Grassland and Forage Science
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• v.24 no.4
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• pp.341-346
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• 2004

To determine lethal temperature of birdsfoot trefoil(BFT) and italian ryegrass(IRG) at heat-stressed conditions, seedlings grown in a small pots fur 4 weeks were subjected to different temperature regimes of heat treatment. No apparent damage was observed BFT and IRG were treated at 45, 50 or $60^{\circ}C$ for 1 h. And also heat treatments at 60, 65 and $70^{\circ}C$ for 1 h, both of them were withered and showed damage symptom on their leaves but it was not lethal conditions for the whole plants. By contrast, most of plants were prominently withered within one day after heat treatment at $80^{\circ}C/60min$. When BFT was exposed to $80^{\circ}C/60$ min, they were died within 6 days but there was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 55 min. IRC was also died within 2 days that exposed to $80^{\circ}C/20$ min but there was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 15 min. These results indicate that heat killing temperatures of BFT and IRG plants are $80^{\circ}C/60$ min an $80^{\circ}C/20$ min respectively. Simple viability assay system established in this study will be useful for selection and characterization of heat-tolerant transgenic BFT and IRG plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.3
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• pp.145-150
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• 2001

This study was conducted to obtain the transformed birdsfoot trefoil (Lotus corniculatus L.) plants with BcHSP17.6 gene using Agrobacterium turnefaciens LBA4404 and we confirmed transformed gene from the regenerated birdsfoot trefoil plants. The expression vector, pBKH4 vector, harboring BcHSP17.6 gene was used for production of transgenic birdsfoot trefoil plants. The callus of birdsfoot trefoil was cocultivated with Agrobacteriurn turnefaciens and transformed calli were selected on kanamycin-containing SH-kc medium to regenerate into plants. The transformed birdsfoot trefoil plants were produced 4 momths after cultivation on BOi2Y medium. The transgenic birdsfoot trefoil plants were analyzed by isolation of genomic DNA and genomic Southern hybridization using a -32P labelled BcHSPl7.6 fragments. (Key words : Birdsfoot trefoil, Transgenic plant. BcHSP17.6 gene, Callus induction, Plant regeneration)