• Journal of Plant Biotechnology
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• v.32 no.2
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• pp.123-128
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• 2005

A series of experiments were performed to establish regeneration system through friable embryogenic callus (FFC) of Lilium longiflorum 'Nellie White'. Only hard and regular callus was induced from bulb scales on medium containing 2.0 mg/L dicamba and $30{\sim}90$ g/L sucrose. The induced hard callus was subcultured on medium with 2.0 mg/L dicamba and 30 g/L sucrose, and used as a material for induction of FEC. In order to induce FEC, induced hard and regular callus was chopped into $1{\sim}2\;mm$ segments, and re-cultured on medium with 2.0 mg/L dicamba and 90 g/L sucrose. FEC was induced from chopped hard calli by the subcultures of two months interval. The induction rate of FEC was enhanced when hard callus was subcultured on same medium. FEC was proliferated more than 5 times on medium with $1.0{\sim}2.0\;mg/L$ dicamba and 90 g/L sucrose. Bulblet differentiation from FEC was very favorable on MS medium supplemented with 0.1 mg/L BA, 1.0 mg/L NAA and 30 g/L maltose, but many differentiated bulblets were changed to vitrificated ones. The differentiation of normal bulblets was most effective on medium containing $0.5{\sim}1.0\%$ activated charcoal and 30 g/L sucrose.

• Journal of Plant Biotechnology
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• v.7 no.4
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• pp.259-265
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• 2005

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

• Journal of Plant Biotechnology
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• v.50
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• pp.11-18
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• 2023

Manihot esculenta Crantz, commonly known as cassava, is a staple aliment that is a significant source of revenue for farmers. The embryogenic callus is crucial in the genetic engineering of various crop species, including cassava. Four cultivar cassava landraces from East Java were assessed for their ability to produce friable embryogenic callus (FEC) for protoplast isolation. In this study, four cassava cultivars; (Kaspro, Kuning, Gajah, and Gendruwo); were used to obtain FEC, which involved the culture of immature leaf lobes (ILLs) and apical buds (ABs) media containing MS supplemented with 33 μM picloram and 2 μM of CuSO₄ (M1) or MS supplemented with 50 μM 2,4-D and 2 μM CuSO₄ (M2). The highest FEC induction efficiency ranged from 72% to 57%, and the highest FEC number ranged from 4.7 to 3.7 with AB explants in media containing MS + 33 μM pilocram and 2 μM CuSO₄ (M1). On the other hand, the efficiency of somatic embryogenesis induction ranged from 67% to 53%, and the number ranged from 4.4 to 3.4. The efficiencies of FEC induction ranged from 48% to 42%, and the number ranged from 3.1 to 2.6 with AB explants in media containing MS + 50 μM 2,4-D and 2 μM CuSO₄ (M2); the efficiency of FEC induction ranged from 56% to 50%, and the value ranged from 3.6 to 2.4 with ILL explants. The FEC induction of the Gendruwo cultivar, which was examined using AB and ILL explants, demonstrated the lowest efficiency. Nevertheless, all four cultivars showed the ability to generate FEC, even though their effectiveness differed depending on the explant genotype and the applied media.

• Korean Journal of Medicinal Crop Science
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• v.15 no.2
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• pp.73-81
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• 2007

An efficient regeneration system was developed using leaf, petiole, and internode explants. Highly embryogenic callus was obtained following cultivation on MS basal nutrient supplemented with 2 $mg/{\ell}$ 2,4-D. Globular, heart, torpedo and cotyledon shaped somatic embryo were produced from the surface of embryogenic callus. Direct shoot regeneration without intermediate callus formation has been achieved on MS medium supplemented NAA and BAP. The percentage of response varies with different concentration of auxin and cytokinin treated individually or in combination. The best shoot regeneration response (54.28%) and number of shoot per explant (12.67) were achieved on the medium supplemented with 0.1 $mg/{\ell}$ NAA and 1 $mg/{\ell}$ BAP. The regenerated shoot transformed into young plant when cultured into elongation and root induction medium. More than 90% of in vitro propagated plants could survive when transferred to the greenhouse for acclimation. This optimized regeneration system can be used for rapid shoot proliferation and genetic transformation.

• Korean Journal of Plant Resources
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• v.24 no.2
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• pp.168-173
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• 2011

This study was carried out to optimize the embryogenic callus induction and plant regeneration from mature seeds of Sorghum bicolor. The effect of growth regulators was investigated on formation of embryogenic callus. The highest frequency of embryogenic callus was observed when the mature seeds were cultured on B5 medium supplemented with 2 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D). The highest frequency of plant regeneration from embryogenic callus was observed on MS medium with 0.5 $mg\;l^{-1}$ 6-benzyl amino purine (BAP) and 0.25 $mg\;l^{-1}$ indole-3-butyric acid (IBA) to optimize the shoot regeneration. High concentration of BAP (1 $mg\;l^{-1}$) supplemented with IBA (0.25 $mg\;l^{-1}$) was effective combination for shoot multiplication. MS medium supplemented with 1 $mg\;l^{-1}$ IBA was found to be the most effective for inducing roots. Normal rooted plantlets were transferred to the greenhouse for hardening with over 90% survival rate. Hence, this reproducible protocol could be useful for mass propagation and genetic transformation of S. bicolor.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.245-249
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• 2003

This study was carried out to establish a regeneration system from leaf explant of purple sweetpotato(Ipomoea batatas L.) The optimal concentrations of plant growth regulators for callus induction and shoot formation were determined. The optimal combination for callus formation was 1$\mu$M 2,4-D 5$\mu$M BM, and highest yield of embryogenic calli were observed on Murashige and Skoog basal medium containing 0.5$\mu$M 2,4-D under light condition after 4weeks of culture. Embryogenec callus was subcultured on medium supplemented with 5$\mu$M ABA for 4 days. Subsequently, regeneration of adventitious shoots occurred when these embryogenic calli were transferred onto medium with 3∼6$\mu$M gibberellic acid. Regenerated shoots were developed into normal plantlets.

• Korean Journal of Plant Resources
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• v.29 no.1
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• pp.143-153
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• 2016

Plant regeneration via organogenesis and somatic embryogenesis was investigated in Korean soybean cultivars including Cheongja 3, Jinpumkong 2, Taekwangkong and Uram. Cotyledon, cotyledon+hypocotyl and hypocotyl segments of 7-day-old seedlings were cultured on MS medium containing various concentration (0, 1, 2 and 4 ㎎/L) of BA and TDZ. The results showed that MS medium supplemented with BA 2.0 ㎎/L yielded the highest shoot formation ratio of 83.3%. In 4 cultivars, Taekwangkong showed the highest ratio of shoot formation. When various sizes of immature cotyledons (S: 1∼ 2 ㎜, M: 3∼5 ㎜, L: 6∼8 ㎜) were tested on MS medium containing 2,4-D 40 ㎎/L for somatic embryogenesis, the optimum size for embryogenic callus induction was 3∼5 ㎜ in length of immature cotyledons. In 4 cultivars, Taekwangkong showed the highest percentage of embryogenic callus induction. The results indicate that Taekwangkong is the best soybean cultivar for plant regeneration via organogenesis and embryogenic callus induction among the 4 cultivars.

• Korean Journal of Plant Resources
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• v.12 no.2
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• pp.107-119
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• 1999

In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

• KSBB Journal
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• v.6 no.2
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• pp.201-205
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• 1991

We have investigated influencing factors on viability of Bletilla striata protoplasts electroporated in the presence of various electrical conditions. Cultures of embryogenic callus and embryogenic cell suspension were established with immature seeds of Bletilla striata. Viabilty of electroporated protoplasts was decreased according to the increaseing of electroporation voltage and capacitance. An optimal condition of electroporation for viable protoplasts was in HBM buffer at $4^{\circ}C$.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.187-193
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• 2005

In an effort to optimize tissue culture conditions for genetic transformation of tall fescue (Festuca arundinacea Schreb.), an efficient plant regeneration system from seed-derived calli was established. MS medium containing 6 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.1 mg/L benzyladenine (BA) were optimal for embryogenic callus formation from mature seed and had a strong effect on successive plant regeneration. The plant regeneration frequency above 50% was observed when embryogenic calli induced in this medium were transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. 'Kentucky-31' showed to have high frequencies of embryogenic callus induction and plant regeneration up to 58.3 and 50%, respectively. Addition of sucrose to the regeneration medium as a carbon source increased regeneration frequency up to 55%. A short tissue culture period and high-frequency regeneration system established in this study will be useful for molecular breeding of tall fescue through genetic transformation.