• Journal of Plant Biotechnology
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• 제30권3호
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• pp.293-299
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• 2003

The effects of low dosage ${\gamma}$-radiation on the cell growth and the formation of shikonin derivatives were investigated in callus cultures of Lithospermum erythrorhizon under different medium and light conditions. Gamma radiation significantly affected the cell growed and formation of shikonin derivatives, depending on the culture conditions. In the cell cultures grown on M-9 medium, 2Gy and 16Gy of ${\gamma}$-radiation increased the calli growth and the formation of shikonin derivatives, respectively under 16hr day light condition. When calli were cultured for 60 days in the dark after irradiation of ${\gamma}$-radiation, cell growth was increased at low dosage of 1Gy and 2Gy in LS medium containing BA 2mg/L and IAA 0.2mg/L. Interestingly, calli grown in M-9 medium by 2Gy irradiation for 60 days significantly stimulated the formation of shikonin derivatives(13.21mg/g cell fresh wt), which was approximately 6 times higher than untreated cells.

• Archives of Pharmacal Research
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• 제16권3호
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• pp.244-250
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• 1993

Calli and suspension cultures were obtained following inoculation of the explant from leaves of Ginkgo biloba L on the supplemented MS basal medium. The obtained calli and suspension cultured cells were able to produce detectable amounts of ginkgolides which are known as natural specific PAF antagonists. The production of ginkgolides in the calli and suspension cultured celles were identified using GC/MS, GC and HPLC with authentic ocmpounds. Since the production of ginkgolides A and B the calli and suspension cultured cells had been confirmed, effects of types and concentration of plant growth regulators, media and illumination on the induction and growth of the callus were studied. The concentrations of growth regulators for optimal callus were studied. The concentrations of growth regulators for optimal callus induction were studied. The concentrations of growth regulators for optimal callus induction were 1.0 to 2.0 mg/L for NAA and o.1 mg/L for kinetin. The growth of the Callus seemed to be more simnultaed with the combination of NAA and kinetin than NAA and BA with illumination at all concentration ranges of 1.0 to 4.0 mg/l for NAA and o.1 to 1.0 mg/L for kinetin or BA studied. Amogn 8 different media used, the induction rate of callus on Anderson, Eriksson, and Shenk and Hildebrant at 4 weeks after the innoculation was almost the same as that of MS. However, callus was rarely induced on Heller or White medium. Suspension cultures were easily initiated with 3 g of callus (fresh weight) derived from ginkgo leaves on supplemented MS medium. A typical growth curve of suspension cultured cells could be obtained by measuring the fresh weight of the suspension cultured cells at every 3 days. To improve the growth of suspension cultured cells of ginkgo, effects of concentrations of NAA, sucrose, phosphate ions and molar ratio of $NH_{4}^+\;to\;NO_{3}^-$ ions in the culture medium were studied. The maximum growth of the cells was achieved when the culture medium contained 1.0 mg/L of NAA, 30 g/L sucrose, 1.75 mM phosphate ions and 1:5 molar ratio of $NH_{4}\;to\;NO_{3}^-$ ions.

• Applied Biological Chemistry
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• 제34권2호
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• pp.142-148
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• 1991

개화후 15일된 대두미숙종자를 배양하였을 때 callus의 형성에 적당한 온도는 $24{\sim}27^{\circ}C$였으며, NAA를 생장조절제로 사용하였을 때 embryogenic callus(EC)가 많이 형성되었다. 또한 EC를 계대배양하였을 때 BA 2mg/1 사용한 배지에서는 shoot가, IAA 2mg/1 사용한 배지에서는 root가 많이 형성되었으나, BA 2mg/1 IAA 2mg/1를 동시에 사용한 배지에서는 callus의 형성만 왕성할 뿐, 기관분화의 기미는 보이지 않았다. 배양체에 함유되어 있는 total lipid는 온도가 낮은 배양조건일수록 많았으며, callus의 생육이 왕성한 $24{\sim}27^{\circ}C$에서 glycolipid, phospholipid, free sterol의 함량이 다소 많아졌으며, embryogenic structure가 많이 생겨날 때에는 free sterol의 함량이 많았다. 배양체의 지방산 조성은 EC에서 불포화 지방산의 함량이 많았으며, 불포화도는 0.837이었다. 또한 sterol조성에서도 EC에서 cholesterol의 함량이 다른 배양체에서보다 현저히 높았다.

• Journal of Plant Biotechnology
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• 제5권3호
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• pp.181-185
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• 2003

Callus and suspension cultures derived from leaf explants of Plumbago rosea were established on Murashige and Skoog's medium containing 1 mg/L IAA, 0.5 mg/L NAA and 0.3 mg/L BAP. Callus cultures were tested for their growth and accumulation of plumbagin, a naphthoquinone and was identified by $^1H$ NMR and electron ionization mass spectroscopy. While auxins (not 2,4-D) influenced growth and plumbagin accumulation, cytokinins did not influence them much. Increasing concentrations of IAA in presence of NAA and BAP increased plumbagin in suspensions only up to 1 mg/L. Growth of callus was optimum (8.3 g DCW/I) at a hormonal combination of 1.5 mg/L IAA, 0.5 mg/L NAA and 0.3 mg/L BAP, but high plumbagin accumulation (4.9 mg/g DCW) was recorded at 1.0 mg/L IAA plus 0.3 mg/L BAP. Since instability in growth and secondary metabolite accumulation was noticed, several cell lines/clumps of callus were screened for plumbagin accumulation by visual and analytical methods. Biomass and accumulation of plumbagin showed a negative correlation in several cell lines. But one cell line showed stability both in terms of biomass and plumbagin accumulation over a period of 6 months.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권1호
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• pp.41-46
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• 2004

The influence of various culture conditions on growth and ginkgolides (GKA and GKB), and bilobalide formation in callus and suspension cultures of Ginkgo biloba were investigated. Callus induced from the leaf petioles exhibited distinct morphological and physiological responses. The cell biomass and ginkgolides content varied among the cell lines; brownish callus lines produced high levels of ginkgolides and bilobalide in spite of poor cell growth. Among the culture media used, MS medium showed significant effect on cell growth and ginkgolides production. Low concentration of sucrose (3%) improved cell growth, while higher sucrose levels (5 and 7%) improved ginkgolides production. Cultivation of callus cultures above 28$^{\circ}C$ dramatically reduced their growth rate; however the cell lines grown at 36$^{\circ}C$ showed increased levels of bilobalide content. A 2.5-L balloon type bubble bioreactor (BTBB) was successfully developed for the cell growth and ginkgolides production.

• 한국응용과학기술학회지
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• 제36권4호
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• pp.1485-1495
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• 2019

본 연구는 Trapa Japonica를 껍질을 벗긴 알맹이와 껍질, 그리고 알맹이와 껍질, 캘러스배양물을 추출한 추출물의 항산화, 항염증 효과를 평가하였다. RAW 264.7 세포와 HaCaT 세포를 사용하여 조사한 각각의 독성은 껍질 추출물에서 100 ㎍/㎖의 농도에서 각각 45.8 ± 1.5 %와 51.1 ± 1.0 %의 세포 생존률을 보였으며 그 이상의 농도에서 더 낮은 세포 생존률을 나타내었다. 총폴리페놀 함량은 껍질과 캘러스배양 추출물에서 각각 213.20 ± 15.78 mg/g, 205.20 ± 18.97 mg/g의 함량을 보였으며 플라보노이드 함량도 껍질 추출물은 29.30 ± 3.24mg/g, 캘러스배양 추출물은 37.4 ± 7.43 mg/g으로 다른 추출물에 비해 높은 함량을 나타내었다. 세포 독성은 알맹이 추출물과 캘러스배양 추출물에서는 세포 독성이 거의 나타나지 않았다. 항산화 활성을 확인하기 위한 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) 라디컬 감소는 추출물 농도 0.005 ~ 1000 ㎍/㎖ 범위에서 껍질 추출물 67.53 ± 1.5 % ~ 75.75 ± 0.5 %의 소거능을 보였으며 캘러스배양 추출물은 3.1 ± 0.1 % ~ 77.32 ± 0.5 %의 감소효과를 보였다. 모든 추출물을 6.25, 12.5, 25, 50 ㎍/㎖ 농도에서 LPS로 유도된 Nitric Oxide (NO) 생성량을 측정한 결과 유의한 (p < 0.05, p < 0.01)의 감소가 나타났다.

• 식물조직배양학회지
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• 제21권6호
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• pp.341-345
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• 1994

식물세포의 기내배양에 의한 ubiquinone 10의 생산연구의 일환으로 Nicotiana tabacum cv Xanthi 품종으로부터 ubiquinone 10의 생성여부을 확인하고, 식물호르몬의 종류와 농도의 조건에 따른 ubiquinone 10의 생산증대를 조사하고자 본 실험을 수행하였다. Xanthi 캘러스 생장은 NAA 및 2,4-D와 혼합처리시 생장이 양호하였으며 NAA 단독처리구에서도 생장량이 증가되었고 암배양보다는 광배양상태에서 더 양호하였다. 또한 HPLC에 의해서 ubiquinone 10의 생성 여부를 조사한 결과 어느 조건에서도 모두 ubiquinone 10이 검출되었다. 식물호르몬에 의한 ubiquinone 10의 함량은 2,4-D와 NAA 혼합처리구에서보다 NAA 0.5 mg/L 단독처리구에서 ubiquinone 10의 함량이 증가되었으며, 특히 IBA 1mg/L와 NAA 0.5 mg/L 혼합처리구에서 더 양호한 경향을 보였고 단위 플라스크당 ubiquinone 10의 생산량도 더 높았다. 그러나 2,4-D 0.5 mg/L와 kinetin 0.5 mg/L 혼합한 처리구에서는 캘러스의 생장량은 비록 적었지만 ubiquinone 10의 함량이 가장 높아 단위 플라스크당 생산량은 제일 높은 경향을 보였다.

• Journal of Plant Biotechnology
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• 제39권3호
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• pp.146-153
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• 2012

In this study, the antioxidant property of leaf and callus extracts of five selected in vitro grown Ocimum species (Ocimum sanctum, Ocimum kilimandscharicum, Ocimum gratissimum, Ocimum basilicum, and Ocimum americanum) and their respective callus extracts was investigated. The callus cultures were successfully initiated on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (1mg L) combined with different concentrations (0.1-0.4 mg L) of kinetin as plant growth regulators. Total phenolic contents were estimated using the Folin-Ciocalteu reagent. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power, $Fe^{2+}$ chelating activity, and ${\beta}$-carotenelinoleic acid bleaching assays were used to determine the biological effects of the extracts. Interestingly, all the callus extracts exhibited significant (p<0.05) increase in phenolic contents and antioxidant activity. Furthermore, a liner correlation was obtained between the total phenolic contents and free radical scavenging activity ($R^2$ = 0.783). The extracts of leaves and calluses of Ocimum species exhibited activity in all the in vitro antioxidant assays, but its extent was less potent that the positive controls butylated hydroxyl anisole (BHA) and ascorbic acid. A higher accumulation of phenolics in the callus extracts suggests that isolation of high-concentration materials with antioxidant activivity is possible from in vitro callus cultures rather than field-grown plant organs. Furthermore, these extracts may be used as an effective preservative in the food industry.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.275.1-275
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• 1995

No Abstract, See Full Text

• 농업과학연구
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• 제37권3호
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• pp.373-376
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• 2010

Peppermint (Mentha piperita L.) belongs to a member of the mint family (Lamiaceae) and is widely used in food, cosmetics and medicines. This study was carried to investigate the variation of essential oil content and its composition during callus subculture of M. piperita. For callus induction from the leaf explant of peppermint, the basal medium was supplemented with various concentrations of 2, 4-D. The best callus induction rate (93%) of M. piperita. was obtained in MS medium containing 2 mg/l 2, 4-D. The induced peppermint callus maintained on Lin-Staba medium were studied during a period of 20th subcultures for the stability of essential oil production. Growth rates of peppermint callus increased during prolonged subculture. However, there was a progressive decrease of essential oil content and unstability of monoterpene productions when callus cultures were serially subcultured.