• 한국식품과학회지
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• 제36권1호
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• pp.58-63
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• 2004

수박과 참외의 황용도 증진을 위하여 조직을 개개의 세포로 분리할 수 있는 단세포화 기술을 적용하였다. 그 결과 이들을 시판 식물세포분리효소인 Macerozyme A와 Sumyzyme MC로 30분에서 120분간 반응시간별 단세포 함유 반응물의 특성을 조사하였다. 단세포화로 얻을 수 있는 수율, 반응률, 단세포 함유 반응물의 색도, 점도, 총당, 총펙틴, 총폴리페놀, 무기질, 유리 아미노산 함량을 조사하였으며 또한 분리된 단세포의 형태변화를 입도분포로 조사하였다. 전반적으로 효소반응시간이 경과 할수록 반응물의 성분 함량은 증가하였으나 세포벽에 의하여 내부 구성성분이 보호를 받아 안정하게 유지되는 것으로 나타났다. 수박과 참외의 무기질 함량은 주스보다는 단세포 함유 반응물에서 상대적으로 함량이 높게 나타났다. 참외의 경우 무기질인 K의 함량은 효소반응물과 주스에서 각각 240.8 mg%, 172.7 mg%였다. 본 연구 결과는 수박과 참외로부터 효소반응 물의을 이용하여 새로운 형태의 음료 개발을 위한 기초자료로 활용 가능할 것으로 판단되었다.

• Plant Biotechnology Reports
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• 제2권1호
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• pp.87-92
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• 2008

An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

• Journal of Plant Biotechnology
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• 제1권2호
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• pp.117-121
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• 1999

Effects of jasmonic acid treatment, UV-B and white light treatment on the anthocyanin biosynthesis and cell growth were investigated using the cell suspension culture system of grape (Vitis vinifera L.). Cell growth was not affected by white light irradiation, while it was remarkably suppressed by UV-B irradiation from 8 to 32 h. Anthocyanin accumulation dramatically increased after 16 h from irradiation of UV-B. Simultaneous treatment of jasmonic acid and UV-B increased anthocyanin accumulation by 10-fold. The cell division was restored when anthocyanin was abundantly accumulated after 32 h from UV-B irradiation. Optimum concentration of jasmonic acid was found to be 5 uM for maximum accumulation of anthocyanin. Application of jasmonic acid to grape suspension cells rapidly induced the expression of CHS gene after 2 h from treatment and showed maximum level at 32 h. Simultaneous treatment of jasmonic acid and light also induced CHS gene expression after 2 h, but the maximum level of CHS transcript was observed at 16 h with white light and 8 h with UV-B exposure. The synergistical effects could be explained by the defense mechanism that UV irradiation is mediated in part by alterations in JA and its signaling pathway.

• 농업과학연구
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• 제49권4호
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• pp.795-807
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• 2022

The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10^-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD_95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.

• Journal of Plant Biology
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• 제32권4호
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• pp.247-253
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• 1989

Protoplasts were enzymatically isolated from suspension culture of sweet potato. High yields of single protoplasts were produced from nonembryogenic cell aggregates. However, most protoplasts obtained from embryogenic cell clumps were spontaneously fused during enzyme treatment; a small portion of them remained single. Upon transfer to Murashige and Skoog's(MS) liquid medium supplemented with 0.1 mg/1 6-benzyladenine(BA) and 1 mg/12,4-dichlorophenoxyacetic acid(2,4-D), protoplasts from nonembryogenic cell aggregates sustained cell divisions to form cellus. Upon subculture onto MS media with 0.2 mg/12,4-D or without growth regulators, the callus did not give rise to any organs. On the other hand, first cell division of single protoplasts from embryogenic cell clumps was sporadically observed.

• Applied Biological Chemistry
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• 제36권5호
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• pp.358-363
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• 1993

박하(Mentha piperita) 세포현탁배양에(-)-menthol 생합성 중간체를 투여하여 배양된 세포의 대사경로를 연구하였다. (-)-Limonene을 투여 하였을 때 이는 다른 대사물로 변환되지 않는 것으로 관찰 되었다. (+)-Pulegone은 (+)-isomenthone 및 (-)-menthone으로 변환되었으며, (-)-menthone은 (-)-menthol로 변환되었다. 이 실험은 현탁배양세포가 대부분의 생합성 활성을 유지하고 있으며 (-)-limonene hydroxylase의 활성이 제한적임을 보여 주었다. (-)-Isopiperitenone을 투여하였을 때는 (+)-pulegone, piperitenone, (-)-7-hydroxyisopiperitenone, (R)- 및 (S)-6-hydroxyisopiperitenone이 생성되었다.

• Journal of Plant Biology
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• 제36권3호
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• pp.211-218
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• 1993

Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

• 식물조직배양학회지
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• 제21권1호
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• pp.47-53
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• 1994

본 실험은 포도와 미국자리공의 세포현탁배양계에서 세포생장과 athocyaninn 및 betacyanin 색소생성에 미치는 광의 영향을 조사하였다. 그 결과, 포도의 세포현탁배양계에서의 세포증식 패턴은 광의 효과가 미약하였으나, 미국자리공의 경우는 암배양에 비하여 광조사구에서 뚜렷한 세포증식 패턴을 나타내었다. 포도세포는 12일간의 배양기간중 한번의 생장 피크를 보여주는데 비하여 미국자리공의 배양세포는 두 번의 증식 피크를 보여 광이 세포의 생장 및 분열 주기를 촉진시킬 뿐만 아니라 색소의 축적도 촉진시키는 것으로 나타났으며, 미국자리공 배양세포는 배양 후 4일째에 액포화에 따른 색소의 축적이 확인되었다. 한편, 색소생성에 미치는 광의 영향은 포도 현탁배양계에서는 배양후 6일째부터 생성되기 시작하여 배양 후 10일 전후에 최고치를 보이는데 비하여, 미국자리공의 현탁배양계에서는 세포증식 패턴에 따라 배양 후 4일째와 8일째에 두 번의 피크를 보였다. 그러나 이들 두 가지 모두 암배양하에서는 거의 색소 생합성이 이루어지지 않고 광상태하에서만 색소의 축적을 볼 수 있었다. 한편, 포도 세포배양계에서는 배양 후 4일째에 처리한 광은 약간의 효과가 있었으나, 배양 후 7일째 12시간 이상의 광에 의하여 색소생성이 크게 촉진되는 것을 알 수 있었다. 미국자리공의 경우에는 배양 후 7일 째보다는 오히려 배양 후 4일째에 광이 더욱 효과적이었다. 이상의 사실은 이들 두 가지 색소의 생합성과정에는 광이 절대적으로 필수적이지만 광을 필요로 하는 시기 및 요구하는 광량 등이 서로 다르다는 것을 시사해 준다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1985년도 워크샵 및 심포지엄 북한산국립공원의 식생
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• pp.23-33
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• 1985

Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

• Journal of Plant Biology
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• 제31권2호
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• pp.91-100
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• 1988

The effect of Ca2+ and polyamines on the activity $\beta$-glucan synthetase II(GSII) related to cell wall synthesis was studied in carrot suspension cultured cells. The activity of GS II is four times higher than that of $\beta$-glucan synthetase I in carrot suspension cultured cells and in vitro expreiment, the activity of GSII was increased in response to increase in concentration of Ca2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca2+ and polyamines, the GSII activity was high at 0.1mM of Ca2+ and 1mM of putrescine. Also, polycationic poly-L-lysine and poly-L-ornithine increased about 50% the GSII activity than that of the control, respectively. These results may imply that Ca2+ and polyamines were related to the enzyme activity as a polycationic nature. In addition, verapamil as the calcium channel blocker and flunarizine as an antagonist of calcium mechanism in cytoplasm decreased GSII activity ramarkably, Ca2+ and calmodulin stimulated GSII activity as Ca2+ of free ion rather than Ca2+ calmodulin complex. The effect of 2,4-D on the GSII activity in culture medium is shown to be low at 0.1mg per liter and GSII activity increased about 30% more than that of the 0.1mg/l at the range of 0.3-1.0mg per litere. Cummulative results suggest that Ca2+ and polyfamines stimulate the cell wall synthesis by means of the enhancement of GSII activity responsible for synthesizing the cell wall components.