• KSBB Journal
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• v.24 no.1
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• pp.59-64
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• 2009

The callus formation from inferior leaf of Aloe saponaria was induced in M & S medium supplemented with 10-30 ${\mu}M$ NAA (${\alpha}$-naphthalene acetic acid) and 3-7 ${\mu}M$ kinetin under incubation in the dark at $25^{\circ}C$ for 6 weeks. The hot water extract ($100^{\circ}C$, 24 hrs) from cultured callus was obtained and the components analysis for the extract were examined to determine the callus can synthesized the bioactive component such as Aloe polysaccharide. The freeze dried extract contained the sugar of 53.2%, protein of 7.3%, ash of 18.5% and water of 21% (w/w). Two fractions (Fr-I and Fr-II) were obtained by Sepharose CL-4B gel permeation chromatography and Fr-I, major fraction was further purified with dialysis. From sugar analysis by TLC and GC, the purified Fr-I fraction consisted of glucose (77.6%), galactose (17.7%), mannose (4.7%, w/w) and uronic acid (trace). The molecular weight of purified Fr-I fraction determined by GPC was about 110 kDa.

• Korean Journal of Plant Resources
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• v.10 no.2
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• pp.107-113
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• 1997

Transformants of White Clover(Trifolium repens L.) were efficiently produced from immature seed derived callus cocultivated with Agrobacterium twnefaciens LBA4404 harboring plant binary vector. pBI121, using acetosyringone. The mean frequencies of transformants on the two kanamycin-containing media were 16 to 19% when the immature seed-derived calli were infected with bacteria cultured in the presence of 100$\mu$M acetosyringone compared with 7% in media without acetosyringone. Transgenic white clover was subject to molecular analysis for integration into plant nuclear genome and expression of $\beta$-glucuronidase(GUS) gene. PCR and Northern blot analyses demonstrated that GUS gene was integrated into white clover nuclear genome and expressed into its mRNA. The expression of GUS gene into its protein was confirmed by spectrophotometric assay of GUS activity.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.311-316
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• 1995

In an attempt to obtain intersubgeneric somatic hybrids of Pelargonium aridum and P.zonale, protoplast isolated from the two species were fused by using polyethylene glycol(PEG) and electorfusion methods. Protoplast were isolated from cotyledon and leaf tissues using MS medium containing 550 mM sucrose, 0.7% cellulase (Onozuka R-10) and 0.4% Macerozyme. The optimum number of protoplasts per mL of culture medium was 6 x 10$^4$. Protoplast fused by the electrofusion method were more active than by PEG method. Heterokaryotically fused protoplasts formed calli when cultured in MS medium containing 550 mM glucose, 1 to 2 mg/L NAA and 0.5 to 1 mg/L BA.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.275-279
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• 2003

Excised cotyledons and embryo axises of zygotic embryos of Rhus vemicifera were cultured on Murashige and Skoog(MS) medium with various concentrations of 2,4-D. About 3-5% of explants produced callus. Embryogenic callus was preferentially induced from basal parts of embryo axis of zygotic embryos seeds when they were cultured without removal of seed coats. Somatic embryos were developed from embryogenic callus in growth regulator-free medium after 2-3 subcultures on medium with 1.0mg/L 2,4-D and these embryos were matured to cotyledonary stage. Plantlets with well-developed shoots and roots from embryos were obtained on $\frac{1}{4}$MS medium with GA$_{3}$. After acclimatization of plantlets on artificial soil, they were exposed to soil pots.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.257-261
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• 2003

This experiment was carried out to investigate the effects of plant growth regulators on the organogenesis from the tissue culture of Arabidopsis thaliana stem, and the origin of the callus development. When the stem segments were cultured on medium with 2mg/L IAA or NAA, adventitious roots and trichomes were differentiated after 11 days of culture. Callus vigorously formed on medium with 2/L2,4 after 7 days of culture, but adventitious roots and trichomes were not differentiated from callus after 10 days of culture. This results suggesting that picloram is very effective auxin for the callus formation and organogenesis. Callus weakly formed on 0.05mg/L kinetin, and formed on combination of auxins(2mg/L) with 0.05mg/L kinetin. But the effect of combination of auxins and kinetin the callus formation was less than 2,4-D or picloram alone. A histological examination of callus formed on picloram showed that phloram showed that phloem parenchyma cells were divided and enlarged after 2 days of culture. Cortex parenchyma cells were divided and meristematic nodules were developed from these cells after 4 days of culture. Finally, callus formed on outside of cortex and epidermis by division of meristematic nodules after 7 days of culture.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.293-299
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• 2003

The effects of low dosage ${\gamma}$-radiation on the cell growth and the formation of shikonin derivatives were investigated in callus cultures of Lithospermum erythrorhizon under different medium and light conditions. Gamma radiation significantly affected the cell growed and formation of shikonin derivatives, depending on the culture conditions. In the cell cultures grown on M-9 medium, 2Gy and 16Gy of ${\gamma}$-radiation increased the calli growth and the formation of shikonin derivatives, respectively under 16hr day light condition. When calli were cultured for 60 days in the dark after irradiation of ${\gamma}$-radiation, cell growth was increased at low dosage of 1Gy and 2Gy in LS medium containing BA 2mg/L and IAA 0.2mg/L. Interestingly, calli grown in M-9 medium by 2Gy irradiation for 60 days significantly stimulated the formation of shikonin derivatives(13.21mg/g cell fresh wt), which was approximately 6 times higher than untreated cells.

• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.95-100
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• 2000

The cytosolic glutathione reductase (GR) gene of Brassica campestris L. was introduced into several Japonica cultivars of rice by Agrobacterium tumefaciens and a large number of transgenic plants were produced. Three-week old calli were co-cultivated with A. tumefaciens strain EHA101 carrying the plasmid pIGR1. The efficiency of transformation was differed from rice cultivars. A Japonica cultivar, 'Daeribbyeo' appeared the highest efficiency (42.5%) of transformation among the four cultivars tested. The addition of acetosyringone (50 $\mu$M) during co-cultivation was a key to successful transformation. Transgene fragments were identified by PCR amplification and further confirmed by Southern blot analysis. Mendelian inheritance of the transgenes was confirmed in T$_1$ progeny.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.165-168
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• 1995

The embryos formed from anther culture of peony exhibited divergent morphologies. The frequency of normal embryos with two cotyledons was about two times higher in the embryos formed through direct embryogenesis than those formed from callus. About 69% of the embryos with two cotyledons converted into normal plants, but convention rate of the abnormal embryos with one and three or four cotyledons was only 4 to 9%. The embryos of hem and bowling pin shape did not undergo development into normal plants.

• Journal of Korean Society of Forest Science
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• v.97 no.4
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• pp.452-457
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• 2008

In order to develop an efficient micropropagation technique effect of plant growth regulators (PGRs) affecting on shoot proliferation from shoot apex in Pyrus ussuriensis was tested. Generally, there was no conspicuous effect on shoot induction by the treatment of PGRs and one or two shoots/explant were induced when cultured on MS medium supplemented with BA and/or BA plus NAA. Both apical shoot necrosis and hyperhydric shoots were observed frequently in multiplied shoots, and callus was formed at the basal part of shoots. About 20% spontaneous rooting was achieved in growing shoots, however the proliferated shoots exhibited poor rooting rate in gelrite supported media. When we tried to ex vitro rooting of the shoot cutting, the shoot cuttings rooted up to 50% with 100 mg/L IBA application. The rooted plantlets grew normally after acclimatization in the greenhouse.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.341-346
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• 2003

In an effort to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), an efficient and high-frequency plant regeneration system from seed-derived calli was established. Embryogenic calli induced on MS medium containing 3mg/L 2,4-D and 0.1mg/L BA had significantly improved regeneration ability. Plant regeneration rate was 92％ when embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among three kinds of medium, MS and N6 medium were optimal for embryogenic callus induction and plant regeneration, respectively. Ho difference in callus induction frequency was observed among four cultivars of orchardgrass, however, "Roughrider" cultivar showed higher regenerability with the frequency of 61％. Addition of maltose to the regeneration medium as a carbon source dramatically increased regeneration frequency up to 69％. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of orchardgrass through genetic transformation.