• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.135-135
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• 2003

본 연구는 체세포 핵이식 복제 수정란의 이식에 의한 고능력 한우를 다량 증식하기 위한 방안을 확립하기 위하여 수행되었다. 본 실험에 공여된 체세포는 육질과 육량 등급이 국내에서 100위 이내의 암소 귀세포를 채취하여 동결 및 계대배양하여 사용하였다. 한편, 핵이식 수정란의 준비를 위하여 도축장에서 채취한 난소에서 난자를 회수하여 22시간 성숙배양 후 난구세포를 제거하고 극체가 존재하는 난자만을 선별하여 recipient cytoplasm으로 이용하였다. 난자의 제핵, 체세포 핵이식, 전기융합 및 활성화 처리는 본 실험실의 방법에 준하여 실시하였으며, 핵이식란은 CR1aa 배양액 내에서 5% $CO_2$, 95% Air 및 39$^{\circ}C$의 기상조건하에서 7일간 배양 후 이식에 이용되었다. 한편, 수란축은 2회 이상 정상 발정주기가 확인된 경산우와 미경산우에 25mg의 PG $F_{2}$$\alpha$/를 투여하여 발정을 유기하거나 자연발정우를 선발하여 수란축으로 이용하였다. 그 결과, 배반포기배를 이식한 경우 14두중 5두에서 임신이 확인되었으며 그중 4두에서 유산되었고, 1두는 임신 6개월령으로 정상 발육되고 있는 것이 확인되었지만 상실배기단계에서 이식된 경우는 임신이 되지 않았다. (중략)

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.207-211
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• 1995

Immature zygotic embryos from immature seeds of Wasabia japonica (cv Dalma) were isolated and cultured on modified MS medium supplemented with 2,4-D, IAA, and BA. Immature zygotic embryos were classified into torpedo shape and cotyledon stage. The highest rates of callus formation were obtained of 1.0mg/L IAA(torpedo stage, 90.0%)and 1.0mg/L 2,4D plus 0.1mg/L BA(cotyledany stage,84.3%). Somatic embryos after 60 days of culture. These numerous somatic embryo could be seperated and subcultured on the same media for further propagation. After 90 days of culture, most somatic embryos were developed well organized embryos which were able to produce into whole plants.

• Korean Journal of Plant Tissue Culture
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• v.24 no.6
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• pp.345-349
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• 1997

Somatic embryos were induced through embryogenic callus derived from immature zygotic embryo culture of native Prunus yedoensis in Mt. Halla and regenerated into plantlets successfully. Embryogenic callus was induced most effectively on MS medium with 1.0 mg/L 2, 4-D and 0.1 mg/L BAP at an efficiency of approximately 60% using 45 day-old zygotic embryos after full blooming. Globular somatic embryos were induced from embryogenic callus on MS medium with 1.0 mg/L 2, 4-D and 0.1 mg/L BAP and these globular embryos developed to heart-shaped and cotyledonary embryos on hormone-free MS medium. Normal somatic embryos germinated 49% on 1/2 MS medium and the plants regenerated from the somatic embryos were morphologically normal.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.42-42
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• 2002

본 연구는 한국 재래산양의 핵이식에 있어서 공여핵은 귀세포를, 수핵난자는 소 및 돼지의 난포란을 이용하여 핵이식을 실시하여 복제수정란의 체외발달율과 이종간의 핵이식 가능성을 검토하였다. 공여핵은 재래산양의 귀세포를 채취하여 10% FBS 가 첨가된 TCM-199 배양액으로 체외 배양을 실시하여 monolayar Confluent 형성후 0.25% Trypsin-EDTA을 처리하여 계대배양을 실시하였다. (중략)

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.77-81
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• 1995

When cultured on MS medium supplemented with 0.5 mg/L NAA alone or 1.0 mg/L 2,4-D and 0.5 mg/L BA, immature zygotic embryos of Stewartia koreana formed embryogenic calli and somatic embryos. In investigate effect of sucrose concentration on somatic embryo development, embryogenic calli were transferred to MS basal medium containing 1.5,3, 6 or 9% sucrose. The greatest frequency of somatic embryos was obtained on medium containing 6% sucrose. However addition of 1.5 or 9% sucrose to medium inhibited somatic embryo germination and development into normal plantlet After 5 weeks of hardening culture on medium containing 6% sucrose, somatic embryos were transferred to half strangth MS medium supplemented with 0.1% charcol, wherein these embryo developed into the normal plantlets.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.231-236
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• 1998

Carrot cotyledon explants cultured on MS medium with 1 mg/L 2,4-D were transferred to a hormone-free solid medium overlaid with filter paper in order to elucidate the effect of simple physical treatment on the development and germination of somatic embryos. Transfer of the explants cultured for one week on MS basal medium overlaid with 3 sheets of filter paper on to MS basal medium increased somatic embryo production 2-39 times over the one week culture on medium without filter paper overlay. Maturation and germination of somatic embryos was more prominent on medium overlaid with filter paper than on medium without filter paper. The explants cultured for one week on filter paper overlay added with liquid medium showed prominent decrease in somatic embryo formation compared to filter paper overlay only. It is suggested that the filter paper overlay affected the moisture environment of the somatic embryos developing on it.

• Journal of Plant Biotechnology
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• v.38 no.4
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• pp.258-262
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• 2011

Embryogenic callus was induced from the cultures of immature embryos of Camellia japonica L. on Murashige & Skoog's (MS) solid medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), and then the embryogenic callus was proliferated on same medium for 4 weeks over. The embryogenic callus was sub-cultured on MS basal medium without 2,4-D to produce coyledonary stage of somatic embryo. The frequency (%) of somatic embryogenesis was 25.1%, and the majority of somatic embryos formed had a abnormal morphology with cupshaped cotyledon (48.3%), one cotyledon (12.6%), three cotyledons (9.4%), four cotyledons (1.9%), whereas was only normal morphology with two cotyledon (27.5%). When the somatic embryos with normal or abnormal cotyledons transfer to MS basal medium or $\frac{1}{2}$ MS medium with/or without plant growth regulators ($GA_3$, IBA) for regeneration, the frequency (%) of two-cotyledon embryos regenerated into plantlets was higher 11.1% than one cotyledon (0.0~8.3 %), three cotyledons (0.0~5.8%), four cotyledons (0.0%), cup-shaped (0.3~4.2%). These results demonstrated that the anomalous cotyledons of somatic embryos could caused to decrease the rate of plant regeneration.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.143-148
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• 1995

The competence of callus formation and plant regeneration from root derived callus was higher in japonica cultivars than those of Tongil-type cultivars of rice. A japonica type cultivars Yeongdeogbyeo, showed the highest capacity (13%) for plant regeneration from root calli of 6 cultivars tested. The callus induced from seed and root tissues maintained higher capacity for plant regeneration during 7 passages of subculture on N$_{6}$ solid media at 2-week intervals. The maximum frequency (2 x 10$^{5}$ mL) of round cells and their cell colonies showed about 24 days after suspension culture of root-derived callus in N$_{6}$ medium with lmg/L 2,4-D, 300mg/L casein hydrolysate, 10mM L-proline, 20g/L sucrose and 30g/L sorbitol. The frequency of somatic embryo formation in suspension cultures of root-derived callus increased with prolonged advance of subculture time from 30 to 90 days, but their regenerative capacities decreased.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.196-204
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• 2012

This study was carried out to develop reliable systems for somatic embryogenesis in oil palm tree (Elaeis guineensis Jacq.), and to verify the somaclonal variants by RAPD analysis. Embryogenic callus was induced successfully on modified half-strength MS medium containing $NaH_2PO_4{\cdot}2H_2O$ and casein. Embryogenic callus was further developed to somatic embryo mass (SEM), which is very hard and bonded tightly each other. Plantlets were proliferated when SEM was cultured on modified MS medium containing half strength $NH_4NO_3$, casein and L-ascorbic acid. Plantlets were transplanted into pots containing artificial soils. When RAPD analysis was conducted using randomly selected 95 in vitro plantlets and 19 random primers, somaclonal variation was detected using BNR35 primer. There was missing band around 1 kb in #22, #28, #35, and #77 plantlets. In addition, bands obtained from #28, #35, and #77 was much stronger than other normal bands. The blast results at NCBI revealed that somaclonal variation observed in this study was related to chloroplast genome of oil palm. The results also revealed that oil palm reproduction system through somatic embryogenesis is quite reliable and early detection of somaclonal variants seem to be possible at in vitro stage by RAPD analysis.

• Korean Journal of Medicinal Crop Science
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• v.13 no.1
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• pp.57-62
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• 2005

Salt-tolerant transgenic Panax ginseng plants were produced by introducing the SAL1 geue (3'(2'), 5'-bis-phosphate nucleotidase) that confers tolerance to the salts through Agrobacterium tumefaciens co-cultivation. Cotyledon explants of immature ginseng zygotic embryos cultured on Murashige and Skoog medium lacking growth regulators formed somatic embryos directly with below 10%, but the 74% tranformation rate were observed at the treatment of phytohormone with 1.0 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos were initially cultured on MS medium supplemented with 250 mg/l cefotaxime for 3 weeks and subsequently subcultured five times to a medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime. Upon development into the cotyledonary stage, these somatic embryos were transferred to on the medium containing 50 mg/l kanamycin and 10 mg/l gibberellic acid to induce germination and strong selection. Integration of the transgene into the plants was confirmed by polymerase chain reaction with specific primers. The ginseng transformants with well-developed shoots and roots were successfully acclimatized in a greenhouse when they were planted in soil.