• Korean Journal of Medicinal Crop Science
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• v.3 no.2
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• pp.100-106
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• 1995

This study was conducted to find the factors affecting somatic embryogenesis in suspension culture of Rehmania glutinosa and investigate the possibility of artificial seed production by encapsulation of somatic embryos. Linsmeier-Skoog medium was appeared as proper for somatic embryogenesis. Sucrose with $3{\sim}5%$ as carbon sources was good for somatic embryogenesis, and both ammonium and nitrate nitrogen were necesary for normal somatic embryo production. BA with NAA or kinetin with NAA were better than the use of cytokinin alone for both somatic embryogenesis and numbers of somatic embryos. $AgNO_3$ as protectant for vitrification of seedlings in vitro culture had no harmful effect on somatic embryos. Sphericity of encapsulated seeds was good at 3% gel of sodium alginate but germination was better at 2.5% sodium alginate level. Artificial seeds were germinated and developed normal shoots and roots under in vitro condition.

• Journal of Plant Biotechnology
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• v.39 no.2
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• pp.93-98
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• 2012

This study was conducted to evaluate effects of somatic embryos (SEs) induction with different kinds and concentrations of osmoticum, with gelrite and SEs germination with different abscisic acid (ABA) and embryogenic tissue lines (ETLs) in Japanese red pine (Pinus densiflora). In comparison of somatic embryos induction with different kinds and concentrations of osmoticum, the highest record (45/90 mg FW) was obtained from the treatment of 0.1 M maltose+3.75% PEG 4000. In addition, the higher one was also recorded from 0.2 M maltose (41 SEs), it turned out this treatment was also effective in induction of SEs with previous one. In effects of various gelrite concentrations for SEs production, no SEs were occurred in the treatment of 0.4 or 0.6% gelrite, however, the highest no. of SEs shown in 1.0% gellrite (41 SEs), and some comparable results were also marked with 0.8 (37.3 SEs) or 1.2% (39.7 SEs) gelrite. Therefore, SEs can be produced from above 0.8% concentration of gelrite. Finally, in comparison of SEs germination with different concentrations of ABA, ETLs, and AC, the best germination rates (45%) were obtained from the SEs derived from both concentrations of 150 and $200{\mu}M$ ABA in 06-6 ETL and when cultured on germination supplemented with 0.2% AC. When SEs were cultured on germination medium without AC, best germination rate (28.9%) came from the SEs which cultured on $250{\mu}M$ ABA in 06-6 ETL.

• Proceedings of the Korean Society for Bio-Environment Control Conference
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• 2001.11a
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• pp.174-175
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• 2001

미나리 묘를 대량으로 생산하는데 있어서 체세포배를 이용하는 방법이 매우 유리하다. 체세포배는 계절에 영향을 받지 않고 좁은 공간에서 한꺼번에 많은 량의 묘를 생산할 수 있다. 또한 생물반응기 등을 이용하여 대량 생산할 수 있으면 실생묘를 대체할 수 있을 것으로 생각된다. 배양기 내의 공기 환경의 영향은 삼각플라스크를 이용한 소규모 배양에서는 문제가 되지 않았지만 배양기가 커짐에 따라 공기 환경의 변화도 커져 체세포배의 발생과 발아에 영향을 미친다. 체세포배를 배양하는 기간은 배발생과 발아로 나누어, 세포에서 배가 발생하는 처음 2주 동안에는 밀폐 환경, 즉 환기가 거의 되지 않는 환경이 유리하지만 이후의 발아 동안에는 산소를 많이 요구한다. 삼각플라스크를 이용한 소규모 배양에서는 aluminum foil을 플라스크의 뚜껑으로 하여, 배지를 교체할 때를 제외하고는 배발생과 발아가 진행되는 전 배양 기간 동안 이를 유지한다. 배가 발생하는 초기 2주의 경우는 이 밀폐 환경이 발아를 촉진시킨다. 그러나 배가 발아하여 정상 식물체로 발달하는데는 유리하지 않다. 체세포배는 발아할 때 산소를 많이 필요로 하며 에틸렌이 많이 축적되면 발아율이 낮아지며, 적절한 공기 환경이 주어지면 체세포배 발아가 동조화되어 균일한 묘를 얻을 수 있고 수확시기도 예측할 수 있다. 본 실험에서는 배가 발아하는 기간 동안 플라스크 내의 공기 환경을 다르게 하기 위해 플라스크를 막는 뚜껑의 소재를 달리하여 배양한 후 발아율을 측정하였다. 또한 가장 효율적인 환기량을 구명하기 위하여 인위적으로 플라스크에 공기를 넣어 강제 환기시키는 실험을 수행하였다. 미나리 cell은 처음 8일까지는 생장을 하지 않다가 이후 급속히 생장을 시작하여 35일 정도까지는 생장을 하다가 다시 생장이 둔화되었다. 밀폐시킨 삼각플라스크에서 자라는 Cell은 상태도 좋지 않고 전반적인 증식량도 적었다. Cell은 환기정도에 민감한 것으로 판단되며 삼각플라스크에서 약 35일 정도의 생장 주기를 가지는 것으로 사료된다. 배양 3주까지는 플라스틱 뚜껑으로 밀폐시킨 bottle에서 가장 많은 체세포배를 얻었다. Air filter를 달아 2일 마다 신선한 공기를 넣어 주었을 때는 배의 발달이 많이 늦어져 배양 3주째에 다른 처리보다 배의 수가 훨씬 적었다. 체세포배가 발달하는 동안에는 산소를 많이 요구하지 않으나 성숙하는 동안에는 산소를 많이 요구하는 것으로 생각된다.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.77-77
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• 2002

본 연구는 체세포 및 체세포 핵이식란의 염색체 이상에 체세포의 계대의 영향성을 조사하고자 하였다. 한우 성축의 귀 조직으로 얻어진 세포를 공여세포를 체외성숙 후 제핵된 난자에 핵이식을 실시하였으며, 1.9kv/cm, 20$\mu\textrm{s}$/2times의 전기자극으로 융합후 5$\mu\textrm{g}$/ml의 ionomycin에서 4min, 1.9mM 6-DMAP에서 4h동안 배양함으로써 활성화를 유도하였다. 핵이식란은 CRlaa에서 4일간 배양 후 8 -cell단계에서 중기상의 유도를 위하여 상기 배양액 1ml당 0.05$\mu\textrm{g}$ colcemid에서 6-8시간 더 배양하였다. 이후, 6% Fetal bovine serum이 함유된 1% sodium citrate용액에 20분간 저장처리 후, methanol 5 : aceticacid 1 : distilled water 4로 1차, methanol 3: aceticacid 1 로 조성된 2차, methanol 4 : acetic acid 3 : distilled water 1의 3차고정액으로 1분간 재 침지시켰다. 고정 처리가 완료된 slide는 4% Giemsa용액으로 염색한 후 광학현미경 하에서 핵형 양상을 검경하였다. 체세포의 5계대에서는 684개의 spreads를 검경한 결과 염색체 수는 72%가 정상으로 60개이었고, 24%가 60개 이하였으며 4%가 60개 이상을 보였다. 10계대도 5계대와 비슷하여 71%가 정상, 26%가 60개 이하, 3%가 60개 이상이었고, 15계대에서는 55%가 정상이었고, 30%가 60개이하, 15%가 60개 이상을 보였다. 10계대 까지는 mixoploid의 비율의 변화가 없었으나 15계대에서 현저하게 늘어남을 볼 수 있었다. 또한 체외수정란과 핵이식란의 비교에서는 체외수정란은 250개의 spreads를 검경한 결과 염색체 수는 95.6%가 정상으로 60개이었고, 2.0%가 60개 이하, 2.4%가 60개 이상이었으나, 핵이식란은 204개를 검경하여 88%가 정상이었고, 4.9%가 60개이하, 7.1%가 60개 이상을 보임으로써, 핵이식란이 체외수정란에 비하여 염색체 이상의 비율이 높았다. 따라서 계대에 따라 체세포의 염색체이상의 비율이 상대적으로 증가하고, 체세포 핵이식에 따른 염색체 이상이 생길 수 있음을 알 수 있었다. (이 논문은 농림부 연구비에 의해 수행되었음)

• Korean Journal of Medicinal Crop Science
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• v.6 no.4
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• pp.323-327
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• 1998

The contents of saikosaponin a, c and d in the root of somaclones and wild plants of B. falcatum L. were determined. The saikosaponin contents of wild plants collected from Chongson-gun, Kangwon Province and Yongdok-gun, Kyongbuk Province were higher than those of the somaclones derived from 3-month and 2-year aged calli. The mean values of saikosaponin content in 2 year-old somaclones derived from the callus cultured three months were higher than those in 1 year-old somaclones from 2-year aged callus. Variation of saikosaponin content depending on the culture periods was not found. Four plants among 1-year old somaclones showed saikosaponin content twice as much as wild plants had. This result implies the possible use of somaclonal variation for crop improvement.

• Korean Journal of Plant Resources
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• v.26 no.4
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• pp.511-515
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• 2013

Hypocotyls explants of melon seedling were cultured on Murashige and Skoog's (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg/L benzyl aminopurine (BA) for 6 weeks to produce somatic embryos. In somatic embryos produced through intervening bright yellow friable (BYF) from the explants, somatic embryos with two-cotyledon (26%) and horn-type cotyledon (74%) were observed. The procambial strand of cotyledons was originated from circular procambial tissues of lower hypocotyls. The circular procambial independently divided into two procambial strand at the edge of cotyledonary-node, and then connected to each cotyledon to form somatic embryos with two-cotyledon. When cotyledon was horn-type, the circular procambial strand in lower hypocotyls would continuously remain connected to the cotyledon. However, somatic embryos with two or horn type cotyledon formed an abnormal shoot apex without the tunica-corpus structure or dome shape in the inter-cotyledonary area. These results demonstrated that the variation of cotyledon in somatic embryos was closely related to procambial tissue differentiation and shoot apical formation.

• Korean Journal of Medicinal Crop Science
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• v.3 no.1
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• pp.50-55
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• 1995

This study was conducted to determine the optimum conditions for induction and different somatic of somatic embryos as well as germination of encapsulated and stored somatic embryos. Somatic embryos was better formed in 1/2X MS medium than full - strength MS medium. 0.1 to 1.0mg/lBA and kinetin promoted shoot differentiation of somatic embryos. Higher concentration tend to inhibit differentiation. IAA affect positively both root and shoot growth. In vitro germination of somatic embryos encapsulated with 2% alginate matrix containing 1/2 MS nutrient medium and $AgNO_3$ 5mg/l was 86%. Storage of somatic embryos was effecive at $5^{\circ}C$ but the germination rate decreased with longer storage period. RAPD analyses with plants regenerated from the somatic embryos showed DNA polymorphism, indicating abolition of primer binding site by point mutation, deletion, or insertion of certain sequences.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.291-294
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• 1997

The experiments were carried out to determine the optimum conditions for the direct embrogenesis from the cotyledon derived zygotic embryo culture of Paeonia lactiflora Pall. Different peony tissues derived from zygotic embryos were cultured on MS medium with and without 2,4-D. Somatic embryos were formed from the cotyledons cultured on the medium without 2,4-D. The somatic embryogenesis from cotyledons was promoted in the growth regulator-free MS medium containing 1.65~3.3 g/L $NH_4NO_3$ and 30~40 g/L sucrose. The maximum frequency (80.0%) of somatic embryo formation was obtained from the cotyledons excised from zygotic embryos that cultured on MS medium containing 3.3 g/L $NH_4NO_3$. Epicotyl and roots were elongated from a somatic embryo by adding 0.3 mg/L GA$_3$ in the medium or the cold treatment at 4$^{\circ}C$ more than three weeks at 4$^{\circ}C$.

• Journal of Digital Convergence
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• v.15 no.7
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• pp.415-424
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• 2017

The purpose of this study was to examine the relationship among public's risk characteristics, risk severity, risk perception and risk acceptability of human stem cell technology. The subjects were 300 Koreans selected. The data were analyzed by the exploratory factor analysis, confirmatory factor analysis, correlation analysis and structural equation modeing analysis. The results were as followed. First, public's risk characteristics on human stem cell technology influenced positively on risk severity. Second, public's risk characteristics on human stem cell technology influenced positively on risk perception. Third, public's risk severity on human stem cell technology influenced positively on risk perception. Fourth, public's risk characteristics on human stem cell technology influenced negatively on risk acceptability. Fifth, public's risk severity on human stem cell technology influenced not significantly on risk acceptability. Sixth, public's risk perception on human stem cell technology influenced not significantly on risk acceptability. These results will contribute to develop the risk communication strategy on the acceptability of human stem cell technology.

• Korean Journal of Plant Resources
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• v.11 no.1
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• pp.9-14
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• 1998

Two types of somatic embryos were directly induced from the immature zygotic embryos of the wild prunus yedoensis in Mt. Halla after 16 weeks of culture on MS medium supplemented with 0.1mg/L $GA_3$ and 0.1mg/L BAPor 0.5mg/L $GA_3$ and 0.1mg/L BAP. One was normal single embryo with a single basal part. Normal somatic embryos germinated successfully on 1/2 MS medium. However, abnormal nulticotyledonary somatic embryos, formed shoots only on hormone free MS medium and about 80% of shoots rooted on MS medium with 0.5mg/L IBA. The mximum frequency (62.5%) of normal somatic embryos was directly obtained from the zygotic embryo 30 days after full blooming but it was decreased with further maturation.