• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.243-250
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• 2004

In an effort to optimize tissue culture responses of Italian ryegrass(Lolium multiflorum Lam.) for future genetic manipulations to improve forage characteristics, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of three cultivars, 'Jeanne', 'Florida-80' and 'Metro', as explant tissues. For all explants, MS medium containing 5mg/L 2,4-D was optimal for embryogenic callus induction from mature seed and had a strong effect on successive plant regeneration. The optimal concentration of dicamba for the induction of embryogenic callus from mature seeds was 7mg/L. The highest plant regeneration frequency was observed when embryogenic callus was transferred to N6 medium supplemented with 1mg/L 2,4-D and 5mg/L BA. Plant regeneration frequency of callus cultured in the dark was higher than that of cultured in the light. Casein hydrolysate and L-proline improved both in embryogenic callus induction from mature seeds and plant regeneration. High-frequency regeneration system established in this study will be useful for molecular breeding of Italian ryegrass through genetic transformation.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.388-395
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• 2015

Somatic embryogenesis is as an excellent technology for potential use in plant mass production, germplasm conservation, or genetic engineering. We examined the effect of cold storage using 3 embryogenic callus lines with different levels of embryogenesis competence derived from immature zygotic embryo cultures of Kalopanax setemlobus. Somatic embryo induction, germination and plant conversion were evaluated after 1, 3 and 6 months storage at $4^{\circ}C$ in the dark. Most cold-stored embryogenic calli formed somatic embryos normally even after 6 months; however, the induction rate was gradually decreased by increasing the storage period. The most competent line tended to show a slight decline in somatic embryo induction rate, as compared with other lines after cold storage. In general, cold storage resulted in reduced somatic embryo germination and plant regeneration, although 93% somatic embryo germination and 91% plant conversion were achieved regardless of the storage period. Cold storage led to cell browning and degradation. Additionally, the cell structures were confirmed by the aceto-carmine and evans blue dye evaluation. Collectively, our results showed that embryogenic callus of K. septemlobus could be preserved at $4^{\circ}C$ without subculture for 6 months, and suggested the need for storage of relatively more competent embryogenic calli lines to support somatic embryo induction.

• Plant Biotechnology Reports
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• v.3 no.3
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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.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.170-179
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• 2017

Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

• Korean Journal of Plant Resources
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• v.10 no.2
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• pp.122-127
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• 1997

This study was conducted to establish mass propagation system from the tissue culture using immature embroys in Eleutherococus senticosus. Immature embroys from seeds were removed under the microscope and placed on modified SH and WPM medium containing several plant growth regulators. The calli were well formed on media containing 1mg/l of 2,4-D on modified SH medium and 1mg/l of 2,4-D and 3mg/l of TDZ on WPM medium. Shoot regeneration was better on modified SH or WPM medium with combination of high concentration of TDZ and low concentration of 2,4-D. Treatment of 2,4-D alone was better than treatment of TDZ alone in callus induction on modified SH medium but plant regeneration reversed. Treatment of 2.4-D and TDZ combination was better than treatment of 2,4-D alone in callus induction on WPM medium. The results of callus formation and shoot regeneration on WPM media differed to those of SH media. The rate of callus formation was nearly 83% when 2,4-D was added to SH medium on concentration of 1mg/l. The rate of callus formation was nearly 38% when combination treatment of 2,4-D 1mg/l and TDZ 3mg/l was added to WPM medium. Also, plant regeneration differed depending on the mature degree of immature embryo.

• Journal of Plant Biology
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• v.39 no.4
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• pp.249-256
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• 1996

Beet curly top virus is the DNA virus that is providing useful for basic studies of the infection of Arabidopsis thaliana with viral host and provides a system for studying both resistance and the molecular basis of symptom development. An importnat aspect of symptom development observed in BCTV-infected A. thaliana (ecotype Sei-O) was the induction of cell division on phloem and surrounding cortex cells. Analysis of the expression of GUS reporter gene activity in transgenic plants containing constructs with promoter of the auxin-inducible saur gene showed that saur promoter activity was induced concomitantly in symptomatic tissues at the inflorescence shoot tips of the transgenic lines. The auxin sensitivity tests showed that hypersusceptible ecotype, Sei-O produced more amounts of callus than susceptible ecotype, Col-O. These studies indicated that changes in auxin concentration were involved in the induction of cell division in BCTV-infected plants and clearly demonstrated that there was a strong correlation between auxin-induced gene expression and the activation of cell division.

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.247-252
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• 2004

In vitro cultures of watermelon were treated with antimitotic agent colchicine to induce ploidy alterations, particularly the induction of tetraploids. Explants cotyledon, embryonic end of seed, transverse sections of epicotyl and hypocotyl were cultured on MS media supplemented with BA ($1{\mu}M$) and colchicine ($0.01\%,\;0.05\%\;and\;0.1\%$). Explants were subcultured on colchicine free media after 4 and 7 days. Colchicine had negative effect on in vitro regeneration but this exhibited explants related response. However, hypocotyl section of seedlings induced maximum callus on $0.01\%$ colchicine. Shoot proliferation was more in cotyledon explants cultured on colchicine ($0.01\%$) for four days. Maximum root induction and root number were recorded in embryonic end explants. Overall, cotyledon and embryonic end explants, and low colchicine concentration ($0.01\%$) was found optimal in watermelon regeneration.

• Plant Resources
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• v.4 no.1
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• pp.53-58
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• 2001

The effect of carbenicillin on the dedifferentiation and the regeneration efficiency of plant tissues of horseradish(Armoracia rusticana) was evaluated, Inhibition effect for callus initiation was observed when leaf blade, root and petiole segments were grown on MS medium containing 500 mg/L to 2000 mg/L carbenicillin and 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The regeneration of horseradish shoots from leaf blade, root and petiole explants were decreased as the addition of carbenicillin increased from 1000 mg/L to 2000 mg/L in MS medium containing 0.5 mg/L of 6-benzylaminopurine (BAP) or kinetin. Especially, 500 mg/L carbenicillin treatment significantly inhibited shoot induction when leaf blade explants were grown on hormone-free MS medium. It was suggested that the toxic effects of combinations of carbenicillin and 2,4-D may be due to high auxin activity levels.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.267-270
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• 2003

Oleanolic acid is a triterpenoid compound that exists in Oldenlandia diffusa. Recently, oleanolic acid has been noted for antitumor effect. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the induction of callus and suspension cells. Optimum induction condition for callus and suspension cells of Oldenlandia diffusa was determined to be 0.5 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. The retention time of oleanolic acid was 12.6 min and the specific content of oleanolic acid was 0.41 mg/g dry weight.

• Korean Journal of Plant Tissue Culture
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• v.22 no.1
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• pp.13-17
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• 1995

In order to induce haploid plant through pollen culture, pollens of Paeonia albiflora were cultured on MS liquid medium The development of micospore through pollen culture was examined The effect of low temperature (5$^{\circ}C$, 10 days) pretreatment on callus induction and embryogenesis in pollen culture was not evident Calli derived from pollen gave rise to globular embryos when transferred onto solid medium containing 0.5 mg/, 2,4-L. The effect of low temperature pretreatment and medium. combination to pollen viability was unrecognized. Pollen viability was reduced as the culture proceeded.