• Journal of Plant Biotechnology
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• 제4권3호
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• pp.123-127
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• 2002

Herbicide tolerant rice (Oryza sativa L. cv. Ilpumbyeo) cell lines were selected from $\gamma$-ray-irradiated anther-derived cell cultures. The anther-derived cell clusters were small (300 to 400 ${\mu}{\textrm}{m}$ in diameter) and uniform ones that were screened by miracloth filtering. The cell suspensions were very efficient to plate one layer onto agar medium and to screen target cell lines. Herbicide tolerant cell lines were selected by 5 mg/L cyhalofop butyl (CHB) treatment by using the small cell suspensions on agar N6 medium containing 1 mg/L 2,4-D and 0.2 mg/L kinetin. Of the cell lines, one line (CHB-1) showed stable tolerance at 10 mg/L concentration after 6-month culture without herbicide suspension. Growth stability of CHB-1 was similar to that of control cell line on 10 mg/L CHB containing medium. In this experiment we established herbicide tolerant cell line selection system by using anther-derived uniform-cell suspensions with $\gamma$-ray-irradiation.

• 한국식품영양과학회지
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• 제26권3호
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• pp.430-435
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• 1997

Protopectinases are heterologous group of enzymes that degrades insoluble protopectin which consists of middle lamella between cells of plant tissues. Tissues of potato and carrot could be separated to single cells by addition of protopectinase isolated from Rhizopus sp. Color changes ofthe suspensions treated with protopectinase and mechanically macerated after 2 weeks at 4$^{\circ}C$, were investigated. Color change of the latter was very serious, however, that of the former was insignificant. Furthermore, after heat treatment at 121$^{\circ}C$ for 5min, the constituents of mechanically macerted carrot suspension were separated into two layers, but those of single celled carrot were not. Yields of carrot juices extracted from single celled suspensions and mechanical maceration were 93.6% and 56.0%, respectively. These results support that treatment of protopectinase can increase yield of juices extracted from plant, and manufacture high value-added products in food processing.

• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.673-677
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• 2006

A perfusion culture of transgenic Nicotiana tabacum cell suspensions, transformed to express recombinant glucuronidase (GUS), was successfully performed in a 5-1 stirred tank bioreactor. With 0.1 $day^{-1}$ of perfusion rate, the maximum dry cell weight (DCW) reached to 29.5 g/l in 16 days, which was 2.1-fold higher than the obtained in batch culture (14.3 g/l). In terms of the production of GUS, the volumetric activity could be increased up to 12.8 U/ml by using perfusion, compared with 4.9 U/ml in batch culture. The specific GUS activities in both perfusion and batch cultures were maintained at similar levels, 200-400 U/g DCW. Consequently, a perfusion culture could be a good strategy for the enhanced production of recombinant proteins in a plant cell culture system.

• 한국작물학회지
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• 제42권5호
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• pp.615-625
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• 1997

Indica 벼의 원형질체들로 부터 효과적인 식물체 재분화 방법을 개발하였다. 이 방법은 배형성 진탕 세포 배양체로 부터 나출된 원형질체들을 feeder cell들이 agarose에 embedded된 배지 표면에 놓인 filter membrane 위에 배양하는 것이다. Feeder cell로서 Lolium multiflorum 세포배양체를 사용했을 때가 Oryza ridleyi를 사용했을 때보다 효과적이었고, 원형질체 평탄효율은 세포 배양체 age에 따라 달랐지만 최고로 0.68％ 까지 증가되었다. Carbohydrate source로서 maltose를 사용하거나 maltose와 sucrose를 1:1로 조합했을 때가 sucrose 단독으로 사용했을 때보다 식물체 재분화율이 증가되었고, 고농도의 agarose를 이용하여 원형질체로부터 유도된 캘러스를 dehydration시켰을 때 또한 재분화율이 괄목하게 증가되었다. 식물체 재분화율은 control 캘러스로부터 3.1∼30.6％ 였지만 dehydration처리한 캘러스로부터는 30.7∼70.7％까지 증가되었다. 원형질체로 부터 유도된 식물체들은 형태적으로 정상이며 개화했다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1993년도 학술대회지
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• pp.150-154
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• 1993

• Plant Biotechnology Reports
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• 제2권1호
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• pp.33-39
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• 2008

In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

• 한국약용작물학회지
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• 제15권5호
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• pp.346-351
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• 2007

An efficient somatic embryogenesis and plant regeneration protocol was developed for Schisandra chinensis Baill, using embryogenic cell suspensions and optimized media conditions. Friable embryogenic callus was induced from cotyledonary leaf and hypocotyl explants of 7 days old seedlings on MS agar medium supplemented with 1.0 to $4.0\;mg\;l^{-1}$ of 2,4-dichlorophenoxyacetic acid (2,4-D). Fast growing and well dispersed embryogenic cell suspensions were developed within two months when embryogenic calli were transferred to MS liquid medium containing $1.0\;mg\;l^{-1}\;2,4-D$. One third strength of MS medium was the best for both overall growth and development of somatic embryos in liquid culture. Over 3400 viable somatic embryos were produced from each 150 ml flask with an initial cell density of 30 mg in 30 ml medium. Germinated somatic embryos developed in liquid medium converted into plantlets after transferred to half-strength MS semi-solid medium. Approximately 90% of the converted plantlets were successfully transplanted to soil and grew into fertile plants.

• 한국식물병리학회지
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• 제3권3호
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• pp.180-186
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• 1987

단감나무와 차나무의 표면에서 분리한 Pseudomonas syringae 2계통 PS8401, PS8402를 $2.5\%$ glycerol이 첨가된 nutrient agar에서 배양한 다음, 증류수로 세포현탁액($10^8$ colony forming unit/ml)을 조제하여 마이크로피펫법으로 동결온도를 측정하여 본 결과, 각각 -2.5와 $-3.8^{\circ}C$에서 동결하여 빙핵활성이 인정되었다. 한편 동일한 방법에 의한 증류수의 동결온도는 $-21.8^{\circ}C$였고 옥수수를 비롯한 8가지 작물즙액의 동결온도는 $-11.6^{\circ}C$ 이하였다. PS 8401의 nutrient broth 현탁액$(10^8\;cfu/ml)$을 살포한 옥수수유묘는 $-2^{\circ}C$에서부터 얼기 시작하여 $-4^{\circ}C$가 되면, 거의 전체 식물이 상해를 입었으나 nutrient broth만 살포한 대조구의 유묘는 온도가 $-9^{\circ}C$로 내려가기 전까지는 피해를 입지 않았다. PS8401 현탁액을 살포한 후 48시간에 측정한 유묘의 피해는 살포한 세균의 량이 증가함에 따라 심해지는 경향이었다. 상기 PS 8401의 빙핵활성은 현탁액내 세포의 수가 많아짐에 따라 증가하였으며 세균을 배양한 배지의 조성이 배양온도에 따라서도 빙핵활성이 변하였는데 $2.5\%$ glycerol 혹은 $2.5\%$ glucose를 첨가한 nutrient agar와 탄소원을 별도로 첨가하지 않은 nutrient agar에서 자란 세균현탁액$(10^2\;cfu/ml)$의 동결온도는 각각 -4.0, -4.4, $-7.2^{\circ}C$였고, 배지를 $2.5\%$ glycerol을 함유한 nutrient agar로 고정하고 온도를 달리하여 생육시킨 세균현탁액 $(10^2\;cfu/ml)$의 동결온도는 $-4.0^{\circ}C$(생육온도가 $15\~25^{\circ}C$인 경우)와 $-7.6^{\circ}C$(생육온도가 $30^{\circ}C$인 경우)였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.301-304
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• 2002

참당귀 (Angelica gigas Nakai) 현탁세포의 고농도 배양을 위하여 perfusion 배양을 한 결과 세포의 생장이 증진됨을 확인하였다 . 배양 5 일째부터 sucrose를 4배 농축한 배지로 2 mL 씩 교환해준 결과, 세포의 생장이 계속 유지되었고 최대 세포농도도 23.7 g/L 로 대조구에 비하여 1.7 배 증가하였다. 초기 당농도를 50 g/L 로 하여 배지 교환을 한 결과, 최대 세포농도도 23.8 g/L 로 30 g/L 당농도 대조구에 비하여 1.6 배 증가하였고, 배지교환을 하지 않은 50 g/L 대조구에 비하여 1.1 배 증가하였다 . 따라서 초기 당농도를 높이고 연속적으로 배지교환을 해준다면, 고농도의 세포를 얻을 수 있으며 결과적으로 참당귀 세포가 생산하는 다당류의 생산량을 증진시킬 수 있을 것이다.

• Journal of Plant Biology
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• 제34권1호
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• pp.19-23
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• 1991

벼의 종자로부터 embryogenic callus를 유도하고 이로부터 개체로의 재분화를 유도하였으며, embroyogenic cell suspension culture를 통하여, 이로부터 원형질체를 분리, 배양하여 embryogenic callus를 형성하였다. 또한, 분리된 원형질체를 electroporation하였을 때 생존율(viability)에 미치는 여러 요인들을 조사하였다. 원형질체의 생존율은 voltage와 capactiance가 증가할수록 감소를 보였으며, HBM buffer에서 $4^{\circ}C$로 electroporation 하였을 때 생존율에 보다 효과적이었다.