• 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.

• 한국작물학회지
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• 제29권3호
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• pp.232-241
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• 1984

수도 약배양의 효율을 높이기 위하여 callus 유기와 녹색체 출현율에 영향하는 화분의 발육단계, 화분의 저온처리와 기관 및 배지의 영향에 대한 일련의 실험을 수행하였으며 이에서 얻어진 결과를 요약하면 다음과 같다. 1. 단핵기의 화분에서 callus 유기율이 높았으며 이때는 대체로 엽신간장이 5~8cm에 해당되나 품종에 따라 차이가 있다. 2. 이삭을 상, 중, 하로 나누어 조사한 결과 엽이간장이 5~8cm의 것은 상부와 중간부분에 단핵기에 해당되는 것으로 나타났고 9~10cm 정도의 것은 중부와 하부 부분이 단핵기에 해당하는 것으로 나타났다. 3. 일반적으로 저온처리는 8$^{\circ}C$에서 12$^{\circ}C$가 callus 유기에 알맞는 것으로 나타났다. 그러나 온도에 대한 반응은 품종에 따라 차이가 있어 4$^{\circ}C$에서 callus 유기가 잘된 것도 있었다. 4. 저온 처리기간은 일반적으로 8~12일 정도에서 callus 유기가 잘 되었으나 품종간 차이가 있다. 5. 본 실험에서는 N6D 배지보다 N6 배지에서 callus 유기율이 높았다. 6. 대체로 녹색체 출현율은 4$^{\circ}C$와 8$^{\circ}C$로 처리한 것이 12$^{\circ}C$ 처리보다 높았다. 그러나 품종간 차이가 있었다. 7. 녹색체 출현에 좋은 결과를 주었던 온도 처리기간은 대체로 8~12일로 나타났다. 8. 본 실험에서 녹색체 출현율은 N6 배지보다 개량 MS 배지에서 2배 정도 높았다.

• 한국초지조사료학회지
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• 제21권2호
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• pp.49-52
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• 2001

conditions for callus induction and plant regeneratin from seeds of lawngrass (Zoysia japonica Steud.) were confirmed in this study. MS (Murashige and Skoog) medium containg 2,4-D 3 or 5mg/l was used for callus induction, and MS medium with different volumes of BA, NAA and kinetin hormones was used to regenerate the plants from induced calli. MS basic medium containing agar with no hormones or kinetin 1.0mg/l and MS basic medium containing gelite and NAA 1.0mg/l were higher for green callus induction. MS medium containing agar and kinetin 1.0mg/l ws highest degree of efficiency for plant regeneration.

• Journal of Plant Biotechnology
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• 제49권3호
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• pp.187-192
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• 2022

식물 캘러스의 월간 또는 주간의 반복적인 계대배양은 노동 집약적이며 모 세포주로부터 somaclonal variation 발생 위험을 증가시킨다. 식물 캘러스를 보존하기위한 가장 효과적인 방법은 액체질소에 저장하는 초저온동결보존 방법이다. 하지만 초저온동결보존 방법은 동일한 방법으로 여러 식물의 캘러스에 적용할 수 없어 보존 방법 개발에 많은 어려움이 따른다. 또한 해동 후 냉동되어진 캘러스의 생존과 생존 후 재생 속도가 불확실 하다는 단점이 있다. 그러므로 활성 상태의 식물 캘러스의 계대배양 간격을 증가시키는 방법의 개발이 필요하다. 본 연구에서는 냉동과정 없는 활성상태의 다양한 종의 식물 캘러스를 계대배양 없이 4주에서 12주 동안 15℃에서 배양하였다. 25℃에서 12주간 배양한 8종류의 식물 캘러스들은 모두 2배 이하의 성장을 보였으나 15℃에서 12주간 배양 조건에서는 8종류의 식물 캘러스들은 최소 2배 이상의 성장을 하였다. 또한 15℃에서 배양 후 25℃에서 회복시킨 캘러스들 사이의 항산화 활성 역시 크게 변화하지 변하지 않았다. 이러한 결과는 배지조성이나 특별한 새로운 과정없이 15℃ 저온으로 계대배양 간격을 증가시킬 수 있음을 보여준다. 또한 여러 식물 종의 캘러스들 모두에서 긍정적인 결과는 여러 캘러스들에 동일한 방법으로 계대배양 간격을 증가시킴으로 노동력 감소는 물론 somaclonal variation을 상대적으로 줄여 줄 것으로 예상한다.

• Plant Resources
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• 제5권1호
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• pp.78-85
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• 2002

The material of Rhodiola sachatinensis collected from an alpine region of the west-northern China. For analysing the effect, 1 used Rhodiola sachatinensis's rhizome and cultivated callus. In EtOAc, BuOH, $H_2O$ separation the plant showed strong antioxidative activity, but not in Hexane. The radical scavenging effect of EtOAc(RC$_{50}$,35(g), BuOH(RC$_{50}$, 43(g), H$_2$0(RC$_{50}$, 50(g) fraction and MeOH extract(RC$_{50}$, 50(g) of the Rhodiola sachatinensis was comparable to that of synthetic antioxidant BHA(RC$_{50}$, 14(g) and $\alpha$-Tocopherol(RC$_{50}$, 12(g). Total amino acid concentration of plant of In nature condition were 18,009ppm, and major components were arginine, glutamic acid, aspartic acid and valine. The ratio of essential/total amino acid on plant of In nature condition was 46.93%. Total amino acid concentration of callus of In vitro condition were 32,435ppm, and major components were valine, histidine, lysine and leucine. The ratio of essential/total amino acid on callus of In vitro condition was 56.07%. was 56.07%.

• 한국약용작물학회지
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• 제17권3호
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• pp.192-197
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• 2009

Camptotheca acuminata, a native of South China is a well known natural source of monoterpene-indole alkaloid camptothecin(CPT), one of the most promising anti-tumoural compounds. This study was conducted to optimize plant growth regulators and culture conditions on plantlets regeneration through organogenesis from callus of Camptotheca acuminta. Callus were induced from various explants of in vitro germinated plantlets of C. acuminta using WPM medium containing 0.2 ㎎/L 2,4-D. Hypocotyl segments were exhibited higher embryogenic callus than the other explants. Shoot buds formation from embryogenic callus was affected by plant growth regulators, pre-treated dark condition and liquid culture. Organogenesis was optimal in WPM liquid medium containing 0.5 ㎎/L BA. The dark pre-treatment for 2 weeks before the solid culture was effective for organogenesis. The regenerated shoots were rooted in WPM medium with 0.2 ㎎/L NAA and successfully acclimated in green-house conditions.

• Journal of Plant Biotechnology
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• 제4권1호
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• pp.17-21
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• 2002

To establish an the mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of various combinations of auxins and cytokinins on friable callus production were studied. for friable callus production, 2,4-D was superior to other regulators. IAA at 2 mg/L induced callus from stem and petiole while NAA resulted in rooting. Callus induction rate increased with the 2,4-D level, and stem segments were superior to leaf blade or petiole, showing nearly 100% with 1 and 2 mg/L 2,4-D from petiole and stem. Combined treatments of 2,4-D + kinetin and NAA + BA also yielded friable callus from stem segments. In treatments with 1 mg/L 2,4-D + 1 mg/L kinetin and 1 mg/L NAA + 1 mg/L BA, callus induction rate was nearly 100%. The combination effect of 2,4-D and BA on anthocyanin production was not significant.

• 식물조직배양학회지
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• 제27권2호
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• pp.83-87
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• 2000

벼 소포자 유래의 반수성 세포집단을 기내 돌연변이 유기 연구에 이용하기 위하여, 액체배지를 이용한 약 부유배양에서 캘러스 유도에 미치는 생장조절물질의 영향과 유도된 캘러스의 갈변화에 따른 활력저하를 막기 위하여 배지내 항산화제의 첨가효과를 조사하였다. 약 부유배양을 위한 액체배지는 N6배지에 1 mg/L 2,4-D와 1 mg/L kinetin을 첨가한 배지에 서 캘러스 형성율 48.5%, 녹색체 분화율 6.8%, 총 식물체 분화율 9.0%로 가장 높았다. 약 부유배양에서 항산화제 (antioxidant mixture, Sigma Chemical Co.)의 첨가는 캘러스 형성율에 영향을 미치지 않았으며, 캘러스 갈변방지와 총 식물체 분화율 향상에 유의한 효과를 보였다. 항산화제의 적정농도는 250mg/L 였으며, 캘러스 유도배지와 식물체 분화배지에 모두 첨가하는 것이 가장 좋았다.

• Plant Resources
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• 제2권1호
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• pp.1-9
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• 1999

The callus from the hypocotyl region of immature embryo of Citrus junos Sieb. was efficiently induced in the $\frac{1}{2}$ MS medium containing 45uM BA after 8 weeks culture. The callus was developed into the two callus type, embryogenic callus and nonembryogenic callus, which can be distinguished by visual examination depending on color and appearence. In vitro regeneration of callus established efficiently in the hormone-free MS medium from the embryogenic callus. In order to investigate the physiological changes depending on the developmental stage of embryo, the embryo was formed in the MS medium. The embryogenic and nonembryogenic callus, and the various stages of the somatic embryo were examimed the changes of esterase activity, and their isozyme patterns as well. The protein content and esterase activities was gradually increased on the developmental stages of embryo. Total protein pattern were different by the SDS-PAGE and were appeared strong band of 23 KD in the torpedo stage. The pattern of the esterase isozyme was exhibited a difference between embryogenic callus and nonembryogenic callus. It was appered pI 6.0, 8.0, 8.2 in the embryogenic callus. Also the new band of pI 4.75 was appeared in the cotyledon. These results suggest that the changes of esterase activities and isozyme patterns are importent factor in the differentiation and development of citrus.

• Journal of Plant Biotechnology
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• 제37권1호
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• pp.96-101
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• 2010

Leaf explants of the Solanum hainanense plant, grown in vitro, were cultured in basal Murashige and Skoog (MS) media supplemented with 0.5 mg/L kinetin and 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for callus initiation. For maintenance and proliferation, the callus was cultured on MS medium supplemented with 1 mg/L 6-benzylaminopurine (BAP) and 0.5 mg/L 2,4-D. The glycoalkaloid content in the callus was at its maximum after ten weeks of culture (188.65 mg/g), whereas that of the one-year-old control was 22.22 mg/g in the root and 5.99 mg/g in the stem. The glycoalkaloid extracted from the callus inhibited the activity of collagenase on collagen gel. High performance liquid chromatography (HPLC) analysis showed that biotransformation occurred when a callus was grown on medium supplemented with various carbon sources. These results suggest that callus of S. hainanense is a good material for production of glycoalkaloid.