• 식물조직배양학회지
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• 제27권5호
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• pp.387-393
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• 2000

The process by which Agrobacterium tumefaciens genetically transforms plants involves a complex series of reactions communicated between the pathogen and the plants. To identify plant factors involved in agrobacterium-mediated plant transformation, a large number of T-DNA inserted Arabidopsis thaliana mutant lines were investigated for susceptibility to Agrobacterium infection by using an in vitro root inoculation assay. Based on the phenotype of tumorigenesis, twelve T-DNA inserted Arabidopsis mutants(rat) that were resistant to Agrobacterium transformation were found. Three mutants, rat1, rat3, and rat4 were characterized in detail. They showed low transient GUS activity and very low stable transformation efficiency compared to the wild-type plant. The resistance phenotype of rat1 and rats resulted from decreased attachment of Agrobacterium tumefaciens to inoculated root explants. They may be deficient in plant actors that are necessary for bacterial attachment to plant cells. The disrupted genes in rat1, rat3, and rat4 mutants were coding a arabinogalactan protein, a likely cell wall protein and a cellulose synthase-like protein, respectively.

• Plant Resources
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• 제3권3호
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• pp.219-222
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• 2000

For the purpose of establishing an efficient propagation through airlift bioreactor system of Zingiber of officinale Rosc. Korean native Seosanjong, the effect of different factors and bioreactor on cultured plantlets were investigated. The highest number of plantlets, fresh weight per plant was obtained from explants when cultured in MS liquid medium including 0.3 mg/L NAA and 2.0 mg/L kinetin for 40 days. A 10 L bottle type bubble bioreactor, compared with 250 mL Erlenmeyer flask, was more efficient, producing 4.7 plantlets or from 1.5 to 1.6 times more than did the Erlenmeyer flask. The results demonstrate the rapid mass propagation of airlift bioreactor to produce normal ginger.

• 한국약용작물학회지
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• 제8권2호
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• pp.123-128
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• 2000

지황 기내 식물체의 줄기 조직을 재료로 생물반응기에서 부정아 발생 과정을 통하여 기내 종묘를 생산하기 위해 본 실험을 수행하였다. 공기부양형 생물반응기가 교반형에 생물반응기의비해 신초 형성에 유리하였다. 2.5L 규모의 생물반응기에서 적절한 배양 밀도는 배지 1.5L에 시료 50g(줄기절편 90개)이었고, 이 때 공기의 주입량은 0.5 v.v.m으로 조절하는 것이 효과적이었다. 신초의 생산 효율을 높이기 위해 배지에 pH 완충제인 MES를 첨가한 결과, 생성된 신초의 수가 증가하였다. 유리화 억제제를 5g/1의 농도로 배지에 첨가하였을 경우 신초의 유리화 현상이 뚜렷하게 억제됨과 동시에 신초의 형성도 증가되었다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1993년도 제7회 식물생명공학 심포지움 식물 세포 분화의 분자적 접근 Seventh Symposium on Plant Biotechnology -Approach to Plant Cell Differentiation-
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• pp.1-36
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• 1993

It is possible to regenerate plants from calli, single cells and protoplasts of numerous species via organogenasis or embryogenesis in cell and tissue culture systems. Also such regeneration of plants can directly occur from cells of explants. However certain plant species has not been yet provided cultures suitable for plant regeneration from cells or tissues. For example, we have to confirm the regenerability of plant from cells before preparing transformed cells for application. Even more, it is very important to notice that regenerated plants in cell and tissue cultures often show structural abnormality. The mojority of those plants is functionally disordered and eventually cases degenerated. One of such examples is vitreous plants which are manifested mainly in the leaves and manifesteds to a lesser extent in the stems and roots. Regenerants in suspension cultures show more frequent vitrification than on gelled media so that relative humidity and water potential are the key factors involved in abnormal morphogenesis in vitro. The other is that somatic embryos formed in media containing BAP or high concentration of sucrose show frequently cotyledon aberrancy such as polycotyledon and born type cotyledon. The embryos with aberrant cotyledon of Codonopsis lanceolata could not germinate or regenerate into plants in many cases. In contrast, the polycotyledon embryos of Aralia cordata germinated in higher percentage than two cotyledonary embryos, but horn type cotyledonary embryos rarely germinated. The major cause of poor germination is the abnormal development of plumule apex meristem.

• Journal of Plant Biotechnology
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• 제31권3호
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• pp.249-253
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• 2004

생물반응기의 형태에 따른 인삼부정근의 생장과 ginsen-side의 생산능력에 대하여 조사한 결과, 원형의 상부와 하부에 5 cm의 bubble column을 가진 bulb type bubble bioreactor (BU)에서 건물중은 41.92 g으로 가장 많이 증가하였으며, cylindric tube bioreactor (CT)에서 건물중이 38.55g으로 가장 낮게 나타났다. 이들 두 생물반응기의 초기 kLa 값은 BU 생물반응기에서 6.98 h$^{-1}$로 가장 높게 측정되었고, 반대로 bubble column이 없는 CT 생물반응기에서 5.25 h$^{-1}$으로 가장 낮게 측정되었다. 그러나 이와 같은 초기 kLa값의 차이는 부정근내의 이차대사산물인 ginsenoside의 함량에는 영향을 주지 않는 것으로 나타났다. 또한 BU 생물반응기에서 bubble column의 길이를 기존의 5 cm에서 10 cm로 연장시켰을 때, 초기 kLa값이 6.52 h$^{-1}$에서 7.80 h$^{-1}$로 증가하면서 인삼부정근의 생장을 42.13 g에서 50.30 g으로 약 16% 증가시킨 것으로 나타났다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1057-1061
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• 2001

In this study we explored the possibility of performing nuclear transfer in the domestic cat and assessed the ability of different culture media to support in vitro development of reconstructed cat embryos. Donor somatic cells were derived from cultured cumulus cells or explants of oviduct tissue, and recipient cytoplasts from in vitro matured oocytes. A higher percentage of cleavage (84.6% and 86.5%) and development to the morula stage (35.9% and 44.2%) was found when reconstructed embryos receiving cumulus or oviduct cells were cultured in MK1 medium, compared with those cultured in CR1aa (58.7% and 72.5%, 13.8% and 13.6%, respectively). There was no significant difference between MK1 and CR1aa media with respect to the proportion developing to the blastocyst stage (15.4% and 17.3% vs 6.8% and 8.6%, respectively, p>0.05). There was no significant effect (p>0.05) of donor cell type (cumulus and oviduct cells) on the rates of fusion (65.0% and 52.5%), cleavage (84.6% and 86.5%), development to the morula (35.9% and 44.2%), and blastocyst (15.4% and 17.3%) stages when reconstructed embryos were cultured in MK1 medium. Similar results were found for the reconstructed embryos cultured in CR1aa medium. These results show that culture medium has a significant impact on the early development of reconstructed cat embryos, whereas donor cell type does not have a significant effect.

• 생물환경조절학회지
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• 제14권4호
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• pp.298-301
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• 2005

Potato (Solamum tuberosum 'Dejima') plantlets were investigated on culture type and initial quantity of inoculation in bioreactor and survival rate by hydroponics for mass production. rode stems (1 to 1.5cm in length) of potato plantlets multiplied in vitro were grown for 3 weeks in liquid Murashige and Skoog (MS) medium with sucrose $30 g\; L^{-1}$. When plantlets (80-node inoculation) were raised in 10L balloon type bubble (BB) bioreactor, the healthiest growth of plantlets was obtained from explants cultured in ebb & flow culture with medium supplied periodically 12 times per day. The suitable inoculation quantity of 20L BB bioreactor was 120 pieces of stem segments (mean 2.2g fresh weight) in ebb & flow culture. Number of nodal shoot was eight on the average. In controlled culture room, survival rate of plantlets at 7 days after stem cutting was above $70\%$ when they were acclimatized by hydroponics grown in deep flow and solid medium culture. The highest survival rate of the stem cutting plantlets was in nutrient solution adjusted to EC $1.4dS{\cdot}m^{-1}$. Stem cutting plantlets through one culture could be obtained $670\~900$, when plantlets were grown in ebb & flow culture during 3 weeks using a 20L bioreactor with initial 120 pieces of nodal segments. 11 is possible In do mass production of seedlings cultured in bioreactor and hydroponics.

• Journal of Plant Biotechnology
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• 제35권3호
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• pp.223-228
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• 2008

This study was conducted to investigate an optimal condition for shoot proliferation and regenerate shoots from in vitro leaflet and embryogenic calli from in vitro roots in rose. The effect of BAP on shoot proliferation was somewhat different depending upon genotypes or gelling agents. Leaflets with petiole cut from donor shoots which had been cultured in MS medium supplemented with 0.1 $mg{\cdot}L^{-1}$ NAA for six weeks was effective for regeneration of adventitious buds(ABs) as well as shoot elongation of Rosa hybrida cv. Sweet Pink. Culturing seven leaflet explants per petri plate($100mm{\times}15mm$) was effective for regeneration of ABs. Embryogenesis was shown in the calli induced from roots of Rosa hybrida cv. Sweet Pink cultured in the SH medium supplemented with 11 $mg{\cdot}L^{-1}$ 2, 4-D for four weeks. Color of calli induced from roots was yellow although their color was a little different as type of basal medium.

• Plant Biotechnology Reports
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• 제5권3호
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• pp.255-263
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• 2011

Codonopsis lanceolata (Campanulaceae) has been used in traditional medicines, as its roots contain several kinds of triterpenoid saponin with high medicinal values. In this work, we induced transgenic hairy roots of C. lanceolata and analyzed triterpenoid saponins from the hairy roots and hairy root-derived transgenic plants. Hairy roots were obtained from leaf explants by the transformation of Agrobacterium rhizogenes R1000. Transgenic hairy root lines were confirmed by the transcriptional activities of rolA, B, C, and D genes by RT-PCR. Transgenic root lines actively proliferated on hormone-free medium but not in nontransformed roots. Hairy roots contained richer triterpenoids (lancemaside A, foetidissimoside A, and aster saponin Hb) than nontransformed roots. Transgenic plants were regenerated from the hairy roots via somatic embryogenesis. They showed phenotypic alterations such as shortened shoots and an increased number of axillary buds and adventitious roots. The transgenic plants also contained higher triterpenoid levels than wild-type plants. These results suggest that hairy roots and transgenic plants of C. lanceolata could be used as medicinal materials for the production of triterpene saponins.

• 한국작물학회지
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• 제60권3호
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• pp.394-400
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• 2015

본 연구는 2배체와 4배체 도라지 단백질 발현지도 제작과 도라지의 생리활성에 관여하는 메카니즘을 분자적 수준에서 해석하기 위한 기초자료를 얻고자 실시하였다. 2배체 및 4배체 도라지의 단백질의 발현 양상은 배수성에 따른 특이성은 관찰 할 수 없었으며, 모두 분자량 15~100 kDa 크기, pH 4.0~8.0의 범위에 분포하는 것으로 나타났다. 동정된 39개의 단백질 중 2배체에 비해 4배체에서 2개의 단백질이 up-regulated 되었고, 37개의 단백질이 down-regulated 되었다. 단백질을 기능별로 분류한 결과, 산화환원효소의 활성(oxidoreductase activity)기능을 갖는 단백질이 23.7%의 비율로 가장 많았고 다음은 nucleotide binding 기능의 단백질이 15.8%의 비율로 높았다.