• Plant Biotechnology Reports
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• v.5 no.2
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• pp.157-167
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• 2011

In order to establish a reliable and highly efficient method for genetic transformation of pepper, a monitoring system featuring GFP (green fluorescent protein) as a report marker was applied to Agrobacteriummediated transformation. A callus-induced transformation (CIT) system was used to transform the GFP gene. GFP expression was observed in all tissues of $T_0$, $T_1$ and $T_2$ peppers, constituting the first instance in which the whole pepper plant has exhibited GFP fluorescence. A total of 38 T0 peppers were obtained from 4,200 explants. The transformation rate ranged from 0.47 to 1.83% depending on the genotype, which was higher than that obtained by CIT without the GFP monitoring system. This technique could enhance selection power by monitoring GFP expression at the early stage of callus in vitro. The detection of GFP expression in the callus led to successful identification of the shoot that contained the transgene. Thus, this technique saved lots of time and money for conducting the genetic transformation process of pepper. In addition, a co-transformation technique was applied to the target transgene, CaCS (encoding capsaicinoid synthetase of Capsicum) along with GFP. Paprika varieties were transformed by the CaCS::GFP construct, and GFP expression in callus tissues of paprika was monitored to select the right transformant.

• Korean Journal of Plant Resources
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• v.28 no.4
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• pp.526-532
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• 2015

Scutellaria baicalensis Georgi (SJ) is a perennial plant and its root has been used in oriental traditional medicine for treatment of fever, inflammation, diarrhea and anticancer effect, etc. In this study, plant tissue culture system for SJ was developed. Stem piece of younger plant was optimum explant for callus induction and growth on MS medium supplemented with NAA 1.0 ㎎/L plus BA 0.5 ㎎/L. Plantlet regeneration through callus culture was well on MS medium containing NAA 1.0 mg/L. SJ has been known biologically active substances such as baicalin, baiclein, and wogonin. This study was carried out to examine the effect of plant growth regulators for production of baicalin, baicalein, and wogonin through callus culture. The HPLC pattern of callus extract was identical to that of standard solution, it shows that the callus produced by tissue culture has the same flavonoids composition of SJ. Baicalin, baicalein, and wogonin production was 471.5~52.8 ㎍/g, 137.6~4.0 ㎍/g, and 16.6~1.3 ㎍/g, respectively, on MS media with nine different plant growth regulator combinations. This may indicate that plant tissue culture of SJ possible to produce the biologically active substances effectively

• Journal of Animal Environmental Science
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• v.19 no.1
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• pp.55-62
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• 2013

Teff grass is a warm season C4 annual grass that is used for dry hay, silage and haylage. We have developed a high-frequency plant regeneration system for teff grass via callus culture using mature seeds. It was revealed that mature seeds cultured on MS medium supplemented with 2 mg/l 2,4-D, 0.5 g/L proline, 0.5 g/L casamino acid and 3 g/L Gelrite under light condition produced the highest percentage of callus formation (91.9%). Addition of cytokinins (BA) at 0.0~0.5 mg/L to media containing 2 mg/l 2,4-D enhanced callus growth. The most suitable medium for plant regeneration from dehydrated calli was MS agar medium supplemented with 0.1 mg/l NAA, 1 mg/l BA, 0.5 g/L proline, 0.5 g/L casamino acid 3 g/L Gelrite which induced the highest percentage of calli forming shoots (47.0%). The shoots were rooted at the highest rate (100%) when transferred onto 1/2 MS medium and acclimated in greenhouse conditions.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.3
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• pp.191-198
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• 2005

In order to optimize tissue culture conditions for genetic transformation of orchardgrass, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of 3 cultivars, 'Frode'. 'Roughrider' and 'Frontier' as explant tissues. Callus induction medium containing 3mg/L 2,4-D or 3mg/L dicamba each with 0.1mg/L BA was optimal fer embryogenic callus formation from mature seed and had a strong effect on successive plant regeneration. The regeneration frequency from embryogenic callus among cultivars were descending order of Roughrider > Frode > Frontier. Supplementation of the regeneration media with 10mg/L $AgNO_3$ and 40mg/L cysteine enhanced frequency of plant regeneration. Efficient regeneration system established in this study will be useful fur molecular breeding of orchardgrass through genetic transformation.

• Korean Journal of Medicinal Crop Science
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• v.5 no.1
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• pp.7-13
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• 1997

Calli induced from the leaf segment of Bupleurum falcatum were cultured on Mu-rashige and Skoog's(MS) medium supplemented with 2, 4-D, IBA, IAA and NAA of 0.1 mg/l , The induction of adventitious roots from callus was the best in MS medium supplemented with 0.1 mg/l 2, 4-D and the lateral root was the same. The pretreatment of 0.1 mg/l 2, 4-D for 120 hours was most effective for the formation and grwoth of adventitious roots. The number of adventitious roots per micro callus pre-treated with 0.1 mg/l 2, 4- D was 5. 3 which was the highest level. The callus subcultured for 4 weeks were best for the adventitious root formation. The callus subcultured for more than 4 weeks decreased the adventitious root formation and turned to brown in color.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.3
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• pp.223-228
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• 2011

The present study was conducted to determine the optimum in vitro culture condition for callus induction and plant regeneration from mature seeds of bermudagrass (Cynodon dactylon cv. Common). It was revealed that mature seeds cultured on MS medium supplemented with 2 mg/L 2,4-D, 0.5 g/L proline, 0.5 g/L casamino acid and 3 g/L Gelrite under light condition produced the highest percentage of callus formation (39.2%). The most suitable medium for plant regeneration from dehydrated calli was MS agar medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L BA, 0.5 g/L proline, 0.5 g/L casamino acid 3 g/L Gelrite which induced the highest percentage of calli forming shoots (57.7%). The frequency of callus induction and plant regeneration were the highest on sucrose, followed by maltose. The shoots were rooted at the highest rate (100%) when transferred onto 1/2 MS medium. Regenerated plants were morphologically uniform with normal growth pattern.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.3
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• pp.217-222
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• 2011

Success in molecular breeding for better adapted varieties to environmental stresses depend upon the concerted efforts by various research including tissue culture, transformation, genetics and breeding. In order to optimize tissue culture conditions of Siberian wildrye grass, the effects of plant growth regulators on callus induction and plant regeneration were investigated with mature seeds. The highest callus induction frequency was observed when the mature seeds were cultured on MS medium supplemented with 5 mg/L 2,4-D. The highest plant regeneration frequency was observed when callus was transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots were transplanted to the soil. A short tissue culture period and regeneration system would be beneficial for molecular breeding of Siberian wildrye grass by the production of transgenic plant.

• 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.

• Korean Journal of Plant Resources
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• v.12 no.1
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• pp.20-26
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• 1999

To investigate phytotoxic substances in Equisetum arvense, the aqueous extracts(25 and 8$0^{\circ}C$) or the freeze-drying matter from aqueous extracts($25^{\circ}C$) of E. arvense were tested at different concentrations for biological activities on callus induction and growth, seed germination, seedling growth and antibacterial function. Callus induction and growth of Oryza sativa and Brassica campestris ssp. pekinensis were inhibited by the aqueous extracts at 8$0^{\circ}C$ than at $25^{\circ}C$ and the higher concentrations. Callus growth of four receptor species was inhibited in order of Platycodon grandiflorum, Sesamum indicum, Brassica campestris ssp. pekinensis and Oryza sativa. Seed germination of Ο. sativa, S. indicum, and B. campestris ssp. pekinensis was not affected at low concentration, but it was proportionally inhibited by the higher concentrations. The greatest inhibition of seed germination was 28.3% compared to control, when 2,000$\mu\textrm{g}$/ml of freeze-drying matter was applied to B. campestris ssp. pekinensis. Shoot growth was stimulated by 500$\mu\textrm{g}$/ml of freeze-drying matter, but it was inhibited by the higher concentrations. Root growth was significantly inhibited compare to control at all concentrations. Antibacterial activity of freeze-drying matter didn't showed against Xantomonas oryzae and Eschrichia coli, but a small clear zone was formed by 500$\mu\textrm{g}$ of freeze-drying matter against Erwinia carotovora ssp. carotovora.

• Journal of Life Science
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• v.8 no.1
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• pp.8-13
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• 1998

The maize mitochondrial mutant (NCS2) is derived from homologous recombination between genes encoding NADH dehydrogenase subunit 4 and subunit 6. Plants from mitochondria mutants exhibited severe related growth and development including dwarfism and striping on the leaves. Aborted embryos from NCS2 mutants have been rescued and cultured on the N6 medium supplemented with 2,4-D 1 mg/l. Most calli from NCS2 aborted embryos showed slow growing pattern at first stage. However, upon continuous culturing them on the medium, those were segregated into mutant and normal callus lines. These segregations could be detected by using PCR method with three primers. Such segregation seems to be resulted from the preferential growth of normal cells over the mutant cells on the normal culture condition. Therefore, this method can be used for determining rate of indirect cytoplasmic segregation by estimating amplified band intensities. When NCS2 mutant callus lines cultured on regeneration medium, no adventitious shoot induction was observed. However, callus lines with more mitochondria induced adventitious shoots. These studies suggest that mitochondria NADH-dehydrogenase for electron transport in the inner membrane of mitochondria is essential for the differentiation and development of plants.