• Journal of Plant Biotechnology
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• 제6권1호
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• pp.55-61
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• 2004

Callus induction and somatic embryogenesis are fundamental to cotton tissue culture biotechnology. An efficient protocol for callus induction, somatic embryogenesis and their maturation have been developed to regenerate plantlets from cotton (Gossypium hirsutum L.) variety coker 312. Embryogenic callus was initiated from hypo-cotyl region that was used as an explant at seedling stage when it was about 7-8 days old. Callus induction was achieved through culturing hypocotyls (5-7mm) on $MS_{1a} medium supplemented with 2,4-D (0.1 mg/L) and KT (0.5 mg/L) for six weeks. A friable, colorless, bulky and well proliferating callus becomes greenish with the addition of NAA (2.0 mg/L), ZT (0.1 mg/L) and removal of 2,4-D (M $S_{1b}$) cultured for two weeks then again transferred to $MS_{1a}. 2,4-dichlorophenoxyacetic acid (2,4-D) promoted the proliferation of embryogenic callus, but had a negative effect on the differentiation and germination of somatic embryos. ZT (0.1mg/L) and activated charcoal (2g/L), both hormones play an important role in differentiation and germination of somatic embryos in hypocotyls derived embryogenic callus but in case of cotton, such a capability have been observed on MS medium with 1.92 g/L $KNO_3$, but it is considered to attain somewhat more improvement. High embryogenesis frequency was achieved through nutrient deficient stress treatment. The frequency of globular embryogenesis (two-three folds) was achieved when well proliferating callus was (from $MS_{1a}$ media) cultured on MS (1/5 strength) medium for four weeks. Here the development of anthocyanins is the best indicator for somatic embryogenesis. However, when embryoid callus was cultured on MS (full strength) medium, the globular embryos were developed into normal plantlets immediately. In this procedure 27.49% cotyledenary embryos were developed. Of that 70% cotyledenary embryos were developed not only into normal plantlets but rooted simultaneously, when cultured on MS (with 0.05 mgg/L giberrelic acid) medium. So complete plants could be regenerated through somatic embryogenesis from hypocotyl explants within 6 months.s.

• 한국초지조사료학회지
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• 제18권3호
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• pp.267-272
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• 1998

5품종의 오차드그래스 종자로부터 직접 캘러스를 유도하고, 형성된 캘러스로부터 식물체를 재분화하는 조건을 확립하였다. 공시품종증 합성 19호가 캘러스 유도 및 재분화에서 가장 우수한 것으로 판명되었으며, SH, MS, N6 배지주에서 캘러스 유도시에는 MS 배지가, 재분화시에는 N6 배지가 유리한 것으로 나타났다. 각 단계별 배지중 호르몬 첨가는 캘러스 유도 및 증식시에 dicamba $3\;mg/\;{\ell}$, 뿌리와 잎의 유도시에 NAA $1\;mg/\;{\ell}$와 kinetin $5\;mg/\;{\ell}$, 재분화 완성기에 무첨가한 조건이 가장 효율이 좋았으며, 캘러스로부터 완전한 식물체로 재분화되는데 필요한 시간은 약 50~80일이었다.

• Journal of Plant Biotechnology
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• 제34권3호
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• pp.181-187
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• 2007

Orostachys japonicus A. Berger is a Perennial herbaceous plant which has been traditionally used as an anti-inflammatory agent to treat hepatitis and as an anticancer agent. The objective of this study was 1) to establish and proliferate in vitro plant of O. japonicus 2) to induce indirect somatic embryogenesis from O. japonicus. General calli and embryogenic calli in all ranges of 2,4-D and BA combination, were induced and were best at 22% (embryogenic cell) in 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. Embryogenic cell line was maintained by subculture at 2 week intervals and transferred to solid and liquid medium for embryo formation. In solid medium culture, globular and heart shaped embryos were observed in MS medium containing 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. The number of embryos was 6.5 per 0.5 g cell, and then the immature embryos transferred to MS basal medium for embryo development. In a suspension culture of embryogenic cells, globular and heart shaped embryos were emerged in MS medium supplemented with 3.0 mg/L 2,4-D and 0.3 mg/L BA combination after 10 days of incubation. The embryo formation rate was about 33% by suspension culture. The ratio of embryo germination was 60.9%, on the other side, the root formation rate was 74.3% in 1/2 MS continuously.

• 한국지역사회생활과학회지
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• 제24권1호
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• pp.5-12
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• 2013

Caffeine is an alkaloid of the methylxanthine family known as a central nervous system stimulant, temporarily warding off drowsiness and restoring alertness in humans. There is a recommended upper limits of caffeine for health because a high dose can cause negative effects. Chlorogenic acid is a natural polyphenol compound known to have an antioxidant activity. In this study, the contents of caffeine and chlorogenic acid in coffee beans from different origins(Costa Rica, Indonesia, Vietnam) were determined by using liquid chromatography-tandem mass spectrometry(LC-MS/MS). The experiment offers more selectivity and sensitivity for those compounds compared with conventional methods such as UV/VIS spectrophotometry. The average concentrations of caffeine and chlorogenic acid in coffee beans origined in Costa Rica were 15.05 mg/g and 5.33 mg/g respectively. In the case of coffee beans origined in Indonesia, the average concentrations were 13.10 mg/g for caffeine and 3.75 mg/g for chlorogenic acid. Vietnamese coffee showed that the average concentrations were 17.79 mg/g for caffeine and 1.12 mg/g for chlorogenic acid. This study can contribute to a better understanding of the contents of caffeine and chlorogenic acid in various coffee beans in order to evaluate dietary intake.

• Journal of Plant Biotechnology
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• 제6권2호
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• pp.113-118
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• 2004

Somatic embryos were formed from calli obtained from axillary shoots (raised from nodal segments of glasshouse-grown plants under aseptic conditions), internodal segments (from in vitro-raised plants), and root and coty-ledonary leaf segments (from in vitro-raised seedlings) after 8 weeks of initial culture. Embryo formation was the highest (97.33%) from cotyledonary leaf callus on Mura-shige and Skoog's (MS) medium containing kinetin (KN) (3 mg/L). Somatic embryo induction was lesser with different combinations of auxins while it increased to 100% in internodal segment and cotyledonary leaf calli with 6-benzyladenine (BA) (2mg/L) along with 2,3,5-triiodobenzoic acid (TIBA) (2mg/L). The shoots were induced from somatic embryos raised from root, coty-ledonary leaf and internodal segment calli grown on MS medium containing BA in combination with indole-3-acetic acid (IAA). Maximum of 66.67% cultures formed shoots on MS medium containing BA (1mg/L) in combination with IAA (2mg/L). The shoots raised from somatic embryos were rooted on MS medium supplemented with indole-3-butyric acid (IBA) (2mg/L). The plantlets transferred to the field showed 70% survival rate after one year.

• Plant Resources
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• 제6권2호
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• pp.108-113
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• 2003

This study was carried out to obtain basic information for possibility of oral vaccine in carrot using Agrobacteruim -mediated transformation system. The epitope region of porcine epidemic diarrhea virus (PEDV) spike gene which is classified as a member of the Coronaviridae and causes an acute enteritis in pigs was successfully expressed in carrot (Daucus carota) using the Agrobacterium-mediated transformation system. Hypocotyl segments of in vitro germinated plantlets were infected with Agrobacteriun tumefaciens LBA 4404 harboring PEDV spike gene. Embryogenic callus (EC) was induced on MS selection medium with 1 mg/L 2,4-D, 50 mg/L kanamycin and 300 mg/L cefotaxime after 45 days of culture. Subcultured ECs on MS selection medium without 2,4-D were converted to somatic embryos (SE) of various stage; globular, heart and torpedo stage. Putative transgenic embryos were selected on MS medium with 50 mg/L kanamycin and 300 mg/L cefotaxime. Regenerated plantlets from transformed SE were induced on MS medium containing 50 mg/L kanamycin after 30 days of culture. Genomic PCR confirmed the integration of PEDV spike gene into nuclear genome of carrot and northern blot analysis demonstrated the expression of PEDV spike gene in transgenic carrot.

• 한국산림과학회지
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• 제81권2호
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• pp.183-190
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• 1992

기내배양된 현사시나무의 조직배양세포로부터 안토시아닌 생성과 세포생장에 적합한 요인을 구명하여 차후 기내배양을 통한 2차대사산물생산의 기초자료를 제공하고자 연구를 수행하였다. Callus는 0.5mg/l 2, 4-D, 0.1mg/l BAP을 첨가한 MS배지에서 유발시켜 같은 배지에서 증식하였다. 안토지아닌 생산은 MS 기본배지를 기준으로 하여 질산염은 12.5% 감량시키고 인산염은 400%로 증가시켜 5% sucrose와 1.0mg/l IAA 및 1.0mg/l BAP를 첨가한후 7,000 lux의 연속광 하에서 배양했을 때 가장 높게 나타났다. 그러나 세포의 생장은 MS 기본배지를 기준으로 하여 질산염은 50% 감량시키고 인산염은 400%로 증가시켜 5% sucrose와 0.5mg/l 2, 4-D를 첨가한 배지에서 가장 양호한 결과를 얻었다. 안토시아닌의 동정은 1% 염산-메탄올의 유기용매로 추출하여 정제 후 TLC와 UV spectrophotometer로 확인한 결과 pelargonidin 3-rhamnoside-5-glucose로 추정되었다.

• 식물조직배양학회지
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• 제26권1호
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• pp.15-19
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• 1999

Corydalis remota for. peatinata의 꽃자루에서 캘러스를 유도한 후 MS기본배지에 생장조절물질을 첨가하여 체세포 배발생 및 재분화를 시도하였다. 생장조절 물질에 따른 캘러스 유도율을 비교하면 2,4-D 2.0 mg/L 에 zeatin 2.0 mg/L를 혼용 처리한 배지에서 아주 양호한 캘러스 유도 및 생장율을 나타냈다. 식물생장조절물질 또는 호르몬 첨가에 의한 체세포 배의 발생율은 광조건에서 cytokinin류의 농도가 0.5mg/L 단독 첨가한 배지에서 가장 높았다. 또한 발생된 부정배중 MS기본배지에 zeatin 1.0mg/L을 단독처리한 경우가 가장 쌍자엽형성율이 높은 것으로 나타났다. 또한 2,4-D를 미량 첨가하고 cytokinin류를 혼용첨가한 배지나 BAP를 단독 처리한 배지에서는 소식물체의 배지 접지면인 뿌리부분에 활발한 2차 배발생이 나타났다.

• 한국산림과학회지
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• 제78권4호
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• pp.412-418
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• 1989

가중나무(A. altissima, heaven tree)의 형성층(形成層) 조직(組織)에서 callus을 유도(誘導)하여 식물체(植物體)를 재분화(再分化)시키는데 필요(必要)한 제요인(諸要因)을 조사(調査)하였다. callus 유기(誘起)가 가장 좋은 배지(培地) 조합(組合)은 MS 기본배지(基本培地)에 2, 4-D 1.0 mg/l와 BAP 0.1 mg/l을 첨가(添加)한 조합(組合) 이였으며, 유기(誘起)된 callus의 증식(增殖)도 같은 조합(組合)에서 좋은 결과(結果)를 얻었다. callus에서의 재분화는 MS 기본(基本) 배지(培地)에 2, 4-D 0.01mg/l와 BAP 0.5 mg/l 첨가(添加)한 조합(組合) 배지(培地)에서 평균(平均) 5.0개의 줄기를 얻어서 가장 높은 분화율을 관찰할 수 있었다. 재분화된 줄기를 1/2 MS 기본배지(基本培地)에 이식(移植)하였을때 전부 발근(發根)되었으며, 발근(發根)된 식물체(植物體)는 pot로 이식(移植)하여 활착(活着)시켰다.

• 한국자원식물학회지
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• 제6권2호
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• pp.171-179
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• 1993

This study was conducted to investigate the effect of growth regulators and medium composition on the growth of each stage in apical meristem culture for mass propagation of Zanthoxylum piperitum var. inerme Makino. The source material, shoot tip segments were taken from three-years old graft trees. Apical meristems were cultured in vitro on basal MS, GD, WS, half strength MS(1/2MS) and half strength GD(1/2GD) media supplemented with various concentrations for growth regulators(BA, IBA) and inorganic nutrients. The results summarized are as follows: 1. In culture establishment stage, ratio of culture establishment was 96.7% and the best resuit was obtained using MS medium supplemented with 1.0mg/l BA and 0.2mg/l IBA. 2. In shoot multitication stage, both shoot multiplication and growth were achieved in average 5.6cm. These results were obtained on in MS medium supplemented with 1.0mg/l BA and 0.2mg/l IBA. 3. In roothing stage, phloroglucinol(PG) acted as IBA synergist in root initiation. The most faverable combinations for root development was half-strength MS medium supplemented with 162mg/l PG and 0.2mg/l IBA, and ratio of rooting was 58.0%. 4. In Vitro formed plantlets were transplanted to paper pots in greenhouse with 85% of relative humidity. 96% of survival rate was obtained from artificial soil mix having same volume of sand, vermiculite, peat, and soil.