• 생약학회지
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• 제26권4호
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• pp.419-425
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• 1995

Corydalis remota Fish. ex Max. (Papaveraceae) is a well known medicinal plant being used as analgesics or anticonvulsive in oriental medicine. As the alkaloid content is known to vary depending on the environmental factors, the technology of plant tissue culture can be adopted as source of Corydalis-alkaloids. The present study describes an establishment of tissue cultures of Corydalis which produce alkaloids consistently. Callus were induced from immature seeds of Corydalis remota by placing the seeds on MS static media containing NAA(0.25, 1.0 and 4.0 mg/l, respectively). The combined treatment of NAA(1.0 mg/l) with cytokinin(BAP 0.5 mg/l) improved the induction of callus. TLC scanning data followed by sequential extraction and purification revealed that the induced callus contains a significant amount of alkaloids. Cell suspension cultures were established by transferring the induced callus into the liquid media with the same condition of plant growth regulators as the callus culture.

• 식물조직배양학회지
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• 제22권1호
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• pp.29-33
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• 1995

시호 캘러스로부터 유도된 부정근의 saikosaponin 함량을 분석하였다. 캘러스로부터 부정근 발생은 0.1 mg/L a4-D가 첨가된 MS배지에서 가장 양호하였으며, 배양기간에 따른 부정근 형성은 오래된 캘러스에 비해 4주 배양한 캘러스가 효과적이였다. 캘러스당 부정근 형성율은 캘러스의 크기가 900-1000 $\mu$M에서 가장 높았다. Saikosaponin의 총 함량은 배양기간이 길어짐에 따라 현저히 증가하였다. Saikosaponin a와 d의 함량은 배양 150일에서 g부정근 건물 중량당 각각 2089 $\pm$ 124 $\mu$g과 4778 $\pm$ 214 $\mu$g이였으며 saikosaponin c의 함량은 60일 배양에서 3492 $\pm$ 123 $\mu$g 으로 높았다.

• 한국미생물·생명공학회지
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• 제21권4호
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• pp.329-335
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• 1993

Ascorbate oxidase activity in various cucumber tissue extracts was highest in young fruit peeling. Cucumber callus was induced from young fruit peeling and callus cell lines were selected for more than 7 months, which porduced high levels of ascorbate oxidase and had a high growth rate. Induction of callus was optimized with Linsmaier-Skoog(LS) medium at 25$^{\circ}C$ in dark phase. Ascorbate oxidase activity reached a maximum at 5 days after transfer to LS basal liquid-medium ant then declined. The enzyme activity in callus cells was stimulated by addition of 10${\mu}$M $CuSO_4$ in the early logarithmic phase of growth. And also, adding 10${\mu}$M $CuSO_4$ at 3rd day 7th day of culture period, ascorbate oxidase activity in callus cells was maintained to high level. Maximum yield of ascorbate oxidase was found at the 25th day by flask shaking culture, but three-fold of ascorbate oxidase activity was obtained at the 16th day by jar fermentation.

• 식물조직배양학회지
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• 제26권4호
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• pp.223-227
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• 1999

본 연구는 글라디올러스 캘러스로부터의 재분화 조건을 구명함으로써 캘러스 배양시스템을 확립하는데 기여하고자 하였다. 글라디올러스 캘러스로부터의 유식물체 획득은 캘러스를 2,4-D 무첨가 1/2 MS고체배지에 치상한 후 24시간 일장하에서 15$^{\circ}C$로 배양하였을 때 효과적이었다. 캘러스로부터의 기관형성에 미치는 배양방법의 영향을 검토한 결과, 부정근의 형성은 액체진탕배양이 그리고 부정아의 형성은 액채정치배양이 각각 우수한 것으로 확인되었다. 본 실험의 결과를 통해 글라디올러스 ‘Topaz' 캘러스로부터의 재분화를 위한 최적 배양 조건을 선발하는 한편, 액체진탕배양법에 의해 기관-캘러스 혼합체를 유도할 수 있었으며, 이를 이용하여 기내 유식물체를 대량생산할 수 있는 가능성을 확인할 수 있었다.

• Journal of Plant Biotechnology
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• 제37권2호
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• pp.228-235
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• 2010

The establishment of cell suspension culture and plant regeneration of the halophytic Leymus chinensis (Trin.) are described in this study for the first time. Callus induction solid medium containing Murashige and Shoog (MS) basic salt, $2.0\;mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), and $5.0\;mg\;l^{-1}$ L-glutamic acid with $30.0\;g\;l^{-1}$ sucrose and $4.0\;g\;l^{-1}$ gelrite for solidification induced the highest rate of cell division in Type 1 callus among calli of various types. Liquid medium with the same hormone distribution was therefore, used for cell suspension culture from Type 1 callus. Over a 30 d suspension culture at 100 rpm, great amounts of biomass were accumulated, with 71.07% average daily increment and 22.32-fold total fresh weight increment. Comparison of before and after suspension culture, the distribution of different size callus pieces and the maintenance of callus type were basically unaltered, but a slight increase in relative water contents was observed. To induce the potential of plant regeneration, the directly transferring on plant regeneration solid medium containing MS basic salt, $0.2\;mg\;l^{-1}$ $\alpha$-naphthalene acetic acid (NAA), $2.0\;mg\;l^{-1}$ kinetin (Kn), and $2.0\;g\;l^{-1}$ casamino acid and indirectly transferring were simultaneously performed. Even now growth rates of suspension-derived callus on solid medium were approximately half of those of Type 1 callus, but faster somatic embryogenesis was observed. Rooting of all regenerated shoots was successfully performed on half-strength MS medium. All plants appeared phenotypically normal.

• 생명과학회지
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• 제16권7호
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• pp.1141-1147
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• 2006

녹두 유식물체 줄기를 조직배양하여 캘러스조직을 형성하였고 이로부터 부정아 형성을 유도한 다음, 캘러스 조직과 부정아 분화에 대한 기원을 조직해부학적으로 연구하였다. 녹두 줄기절편으로부터 캘러스조직의 유도는 0.5 mg/L 2,4-D와 1.0 mg/L kinetin이 첨가된 MS배지가 효과적이었고, 캘러스로부터의 부정아 분화에는 0.75 mg/L NAA와 1.5 mg/L kinetin이 첨가된 MS배지가 매우 효과적이어서 약 21%의 기관형성율을 나타내었다. 캘러스 조직을 조직 학적으로 관찰한 결과, 캘러스조직은 유관속 형성층 외측에 새롭게 형성된 분열능력이 있는 캘러스 형성층환인_바깥쪽으로 생장을 함으로써 유도되었다. 부정아는 캘러스 조직의 외부 표면부위에서 기원된 부정아 정단분열조직으로부터 발생되었다. 부정아 정단분열조직은 엽원기를 형성하여 나중에 잎이 발생되었다.

• 환경생물
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• 제22권1호
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• pp.148-152
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• 2004

본 실험은 백두산 자생 참돌꽃의 발아조건과 종자, 유식물 및 뿌리유래 캘러스의 유리 아미노산 조성과 함량을 조사하여 다음과 같은 결과를 얻었다. 뿌리로부터 캘러스 유도는 2,4-D 및 NAA가 첨가된 MS배지에서 유도되었다. 캘러스 형성율은 2,4-D 2.0 mg $L^{-1}$에 kinetin 1.0 mg $L^{-1}$ 혼용 처리구가 NAA 2.0 mg $L^{-1}$에 kinetin 1.0 mg $L^{-1}$ 혼용 처리구보다 캘러스 형성율이 높았다. 유리 아미노산은 종자, 줄기, 뿌리 그리고 캘러스에서 aspartic acid, glutamic acid, serine 및 glycine 등의 25∼26종이 검출되었으며 종자, 줄기 그리고 뿌리부위에서는 lysine이 가장 많은 양을 나타냈으며 캘러스에서는 histidine이 높은 양을 나타냈다. 전체적인 유리아미노산 분포는 lysine, histidine 및 arginine의 함량이 높아 참돌꽃 유식물 및 캘러스의 경우 염기성 아미노산의 분포가 높은 식물임을 알 수 있었다. 또한 총 유리 아미노산 함량은 종자와 뿌리, 줄기에 비해 캘러스에서 신선중 g당 500∼l,000$\mu$mo1e정도 높아 우수한 약리작용을 지니고 있는 참돌꽃의 성분을 배양세포를 통하여 대량생산이 가능함을 시사하고 있다.

• Journal of Crop Science and Biotechnology
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• 제10권2호
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• pp.92-97
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• 2007

Antibiotics used for suppressing Agrobacterium in plant transformation procedure might have negligible effects on plant tissues and regeneration. The effects of antibiotics on growth suppression of Agrobacterium and plant regeneration were investigated for enhancing Agrobacterium-mediated transformation using wheat mature embryos. Antibiotics tested, except carbenicillin, were able to suppress that embryos were coated with a layer of Agrobacterium cells in callus induction medium. Agrobacterium growth was suppressed minimally at 50 mg/l of timentin, while cefotaxime and clavamox were completely suppressed at relative high concentration of 250 mg/l. In the treatment of carbenicillin, initiation of growth suppression of Agrobacterium occurred at 750 mg/l of concentration because Agrobacterium KYRT1 contains the carbenicillin resistant gene. In Agrobacterium inoculation, effects of antibiotics were significantly different on the rate of callus induction and shoot formation. Almost embryos were induced calli at 50 mg/l of timentin whereas callus induction rate was achieved above 90% at 100 mg/l and 250 mg/l of cefotaxime and clavamox, respectively. Shoot formation rate was higher in the treatment of timentin than that of cefotaxime and clavamox at 500 mg/l of concentration, respectively. Timentin can be used as a good antibiotics in Agrobacterium-mediated wheat transformation.

• 아시안잔디학회지
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• 제23권2호
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• pp.345-352
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• 2009

한국들잔디는 운동경기장이나 골프 코스 등에 폭넓게 쓰이는 난지형 잔디이다. 많은 품종은 영양체로 번식한다. 이 연구는 배아세포 유도와 식물체 재생을 위한 적정 배양액 개발과 배양조건을 연구하여, 한국잔디의 육종과 번식을 위한 세포배양 체계 확보를 목적으로 하였다. 그 결과 $Cu^{++}\;2.5mgL^{-1}$를 유도 배양액에 첨가하는 것이 캘러스 유도를 위한 최적 농도 조건 이였음을 보여주었다. Sub-culture의 횟수의 증가는 캘러스의 질을 향상시켰다. 세포 부유배양을 위한 최적 용량은 2.5에서 10mL 사이였으며 본 연구에서 58%의 식물체 재생산비율을 보였다.

• Journal of Plant Biotechnology
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• 제50권
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• pp.11-18
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• 2023

Manihot esculenta Crantz, commonly known as cassava, is a staple aliment that is a significant source of revenue for farmers. The embryogenic callus is crucial in the genetic engineering of various crop species, including cassava. Four cultivar cassava landraces from East Java were assessed for their ability to produce friable embryogenic callus (FEC) for protoplast isolation. In this study, four cassava cultivars; (Kaspro, Kuning, Gajah, and Gendruwo); were used to obtain FEC, which involved the culture of immature leaf lobes (ILLs) and apical buds (ABs) media containing MS supplemented with 33 μM picloram and 2 μM of CuSO₄ (M1) or MS supplemented with 50 μM 2,4-D and 2 μM CuSO₄ (M2). The highest FEC induction efficiency ranged from 72% to 57%, and the highest FEC number ranged from 4.7 to 3.7 with AB explants in media containing MS + 33 μM pilocram and 2 μM CuSO₄ (M1). On the other hand, the efficiency of somatic embryogenesis induction ranged from 67% to 53%, and the number ranged from 4.4 to 3.4. The efficiencies of FEC induction ranged from 48% to 42%, and the number ranged from 3.1 to 2.6 with AB explants in media containing MS + 50 μM 2,4-D and 2 μM CuSO₄ (M2); the efficiency of FEC induction ranged from 56% to 50%, and the value ranged from 3.6 to 2.4 with ILL explants. The FEC induction of the Gendruwo cultivar, which was examined using AB and ILL explants, demonstrated the lowest efficiency. Nevertheless, all four cultivars showed the ability to generate FEC, even though their effectiveness differed depending on the explant genotype and the applied media.