• 아시안잔디학회지
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• 제12권4호
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• pp.195-202
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• 1998

The development of a protocol for high efficiency of embryogenic callus separation, maintenance and plant regeneration from the seeds of zoysiagrass cv. Zenith was studied. Embryogenic callus ratio is absolutely determined by genotype, but by adding high concentration of copper into medium, changing light condition and maintaining callus on initial induction medium for 8∼10 weeks, embryogenic callus can be easily distinguished and its growth can be promoted. There were significant differences among selected callus lines (each from one seed) according to their growth rates and regeneration percentages. Callus pre-treatment with activated charcoal inhibited callus growth, increased the level of precocious germination during culture and promoted shoot cluster formation after transfer to regeneration medium. For long-term callus maintenance, N6AA medium was better than MS medium, because the former inhibited non-embryogenic callus formation and kept vigor of embryogenic callus. The best callus lines Z-(5) has been successfully used for transformation and somaclonal variation selection in our laboratory.

• Journal of Plant Biotechnology
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• 제33권4호
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• pp.257-262
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• 2006

The paper evaluated the behavior of in vitro culture responses from a diverse set of Indica rice (Oryza sativa L.) genotypes. Significant differences were found in embryogenic callus induction frequency, callus growth and plant regeneration frequency when mature embryos of 11 cultivars, breeding lines and land races were compared. Genotype as well as plant growth regulator influenced the plant regeneration frequency. Callus induction frequency was not correlated with callus growth as well as plant regeneration frequency. The regenerated plants could grow to normal, fertile plants after they were successfully established in soil.

• 아시안잔디학회지
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• 제11권4호
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• pp.311-320
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• 1997

This study was carried out to induce and maintain callus from 59 zoysiagrass lines, to know the effective disinfestation method for zoysiagrass stolon as explant and the difference in the response of callus induction among 59 lines, and to investigate the effect of medium, growth regulators, light, temperature, stolon part and internode position on callus induction and emhryogenic callus(E.C.) formation. The treatment of 0.lmg/L $HgCl_2$for 15 min resulted in no contamination and the highest callus induction(46.6%). Callus was induced from the 59 zoysiagrass lines. The callus growth of Z. japonica and Z. sinica was generally better than Z. matrella Ten cell lines whose callus and stolon grow fast in culture and in field, respectively were selected to he used for breeding. Callus induction was the most effective at 2.0mg /L of both 2, 4-D and picloram in MS medium. MS medium was the best for callus induction and growth while LS medium was the best for embryogenic callus and shoot formation. Callus induction and growth was better at 28, 31$^{\circ}C$. than 25$^{\circ}C$. and dark condition was better than light condition in MS me-dium containing 2mg/L 2,4-D. While callus induction was better with node part as explant than with internode part, callus growth and embryogenic callus formation was better with internode part. In 'Japonica 1', the first internode was the most effective in callus induction, but third internode was the best in '$M_2$ X $S_2$'.

• Journal of Plant Biotechnology
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• 제32권1호
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• pp.31-35
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• 2005

양파의 재분화 체계 확립을 위해 성숙된 양파의 종자로부터 성숙배를 적출한 후 2,4-D와 BA의 다양한 농도를 이용하여 체세포배발생 캘러스의 형성율을 조사하였다. 체세포 배발생 캘러스는 쉽게 부서지는 특성을 가지고 있었으며 비체세포배발생 캘러스는 물기를 많이 포함하고 있어 쉽게 구분이 가능하였다. 체세포배발생 캘러스는 picloram $2.5\;\cal{mg/L}$ 또는 $5\;\cal{mg/L}$가 단독으로 포함된 MS배지를 이용하여 $25^{\circ}C$ 암배양 4주 후 가장 높은 유도율을 나타내었다. 그러나 유도된 캘러스를 shoot로 재분화 시킨 결과 2,4-D와 BA $1\;\cal{mg/L}$가 혼합된 처리구에서 가장 높은 shoot 유기율을 나타내었다. 따라서 다른 처리에서 유도된 캘러스를 2,4-D와 BA $1\;\cal{mg/L}$가 포함된 액체배지에 현탁배양을 시도하였다. 현탁배양은 캘러스의 증가뿐만 아니라 캘러스의 cell cycle을 synchronized 함으로서 이후 형질전환이 용이할 것으로 판단되었다. 현탁 배양된 캘러스는 Kinetin $1\;\cal{mg/L}$가 포함된 1/2 MS 배지에서 60일간 광배양 하여 shoot 유기하였으며 유도된 shoot는 NAA $0.5\;\cal{mg/L}$가 포함된 발근배지에서 발근 후 순화하여 완전한 식물체로 재생할 수 있었다.

• 아시안잔디학회지
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• 제12권4호
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• pp.189-194
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• 1998

The effect of proline, glutamine, aspartic acid and their combinations on callus induction and embriogenic callus formation from 3 creeping bentgrass (Agrostis palustris) cv. Regent, Mariner, Cato and 1 colonial bentgrass (Agrostis tenuis) cv. Tiger was estimated in both light and dark condition. The addition of amino acids to the growth medium did not have a significant stimulatory effect on the induction of embryogenic callus, instead, they were inhibitory, particularly at higher concentration (40 mM). But supplement of amino acids at lower concentrations (5 or 10mM) to basal medium was beneficial in inhibiting the formation of hairy outgrowth on the surface of embryogenic callus.

• 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.

• Journal of Plant Biology
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• 제31권1호
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• pp.41-49
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• 1988

Several cultivars of rice were examined for induction of embryogenic callus on a medium containing MS salts, vitamins and 2, 4-D under darkness. Embryogenic callus was obtained from cultivar Cheonma with high ratio and embryo-like structures were formed from the callus on a medium with or without reduced 2, 4-D. Somatic embryoids with a plumule and radicle axis surrounded by a scutellum were observed. These embryoids germinated and produced plantlets in 30 days on the same medium. Protoplasts isolated from an embryogenic cell suspension culture derived from embryogenic callus were cultured either in liquid or in agar medium and protoplast derived cell colonies were obtained in 3-4 weeks.

• 문화기술의 융합
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• 제7권3호
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• pp.605-608
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• 2021

나리는 세계 5대 절화에 속하는 주요 절화 중의 하나이며, 아시아, 유라시아 및 북아메리카 등 다양한 지역에 서식한다. 나리 신품종개발을 위해서는 교배, 돌연변이 및 선발육종 기술 외에 조직배양 기술을 포함하는 생명공학기술 도입을 통한 신품종 육성이 필요한데 조직배양 체계확립은 나리 육종체계에 있어서 필수적인 요소 중 하나이다. 조직배양의 많은 분야 중 배발생캘러스를 이용하여 식물체 재분화 증식 체계확립이 많은 작물에서 연구 중인데 본 연구에서도 배발생캘러스 유도에 사용되는 오옥신 중 picloram을 다양한 농도로 처리하여 최적의 배발생캘러스 유도 농도를 선정하고자 실험을 수행하였다. 실험 결과, 3-4주 후부터 CEC (compact embryogenic callus), FEC (friable embryogenic callus) 및 백색캘러스 이렇게 3가지 형태 캘러스가 발생했는데 1.0 mg/l 처리구에서 CEC와 FEC 모두 높은 효율을 나타내었으나 갈변율이 높은 관계로 0.75 mg/l를 나리 배발생캘러스 유도에 적합한 최적 농도로 선정하였다. 이러한 연구결과는 향후 배발생캘러스를 이용한 나리 식물체 재분화 체계 확립 그리고 우량품종 증식체계 확립에 기여할 것이다.

• 식물조직배양학회지
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• 제28권5호
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• pp.277-281
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• 2001

자생 왕포아풀 (Poa pratensis L.)의 배발생 캘러스 유도 및 식물체 재분화 조건을 규명하였다. 도입된 다섯 품종과 국내에서 수집한 네 가지 생태형의 미성숙 화서와 완숙종자를 sucrose 30 g/L, Phytagel 3 g/L와 2,4-D 1 mg/L가 함유된 MS배지에 치상하여 24$^{\circ}C$로 조절된 항온실에서 배양하면서 캘러스를 유도하였다. 대부분의 배양에서 캘러스가 형성되었으나 배발생 캘러스는 완숙종자를 절편체로 썼을 때보다 미숙화서에서, 온실에서 재배되어 형성된 화서보다는 자연 재배조건에서 형성된 화서에서 배발생 캘러스의 유도율이 높았다. 배발생 캘러스는 85~98%의 효율로 높게 유도되었으며 계대배양 및 액체 진탕배양을 통해 6개월 이상 장기 증식되었으며, 생장조절제가 함유되지 않은 MS 재분화 배지에 이식하면 다수의 식물체로 재분화되었다. 재분화 식물들은 온실재배에서도 정상적으로 생장을 하였다.

• Plant Biotechnology Reports
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• 제3권3호
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• pp.243-250
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• 2009

In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production of ${\beta}-carboline$ alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium supplemented with $5.0{\mu}M$ 6 benzyl adenine (BA) and $2.5{\mu}M$ ${\alpha}-naphthaleneacetic$ acid (NAA). The embryogenic callus was maintained on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was at a maximum ($18.1{\pm}0.9$ per g of callus) on MS medium containing $5.0{\mu}M$ BA and $2.5{\mu}M$ NAA together with $75mg\;1^{-1}$ casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation of ${\beta}-carboline$ alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus accumulated equivalent amounts of harmaline ($66.4{\pm}0.5{\mu}g/g$ dry weight), harmine ($82.7{\pm}0.6{\mu}g/g$ dry weight), and diosgenin ($170.7{\pm}1.0{\mu}g/g$ dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals.