• Asian Journal of Turfgrass Science
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• v.25 no.2
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• pp.184-189
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• 2011

To optimize tissue culture conditions for genetic transformation of Zoysia matrella (L.) Merr., we investigated the effects of different plant growth regulators and medium supplements on plant regeneration using stolon explants excised from mature plant grown in a green house. Plant regeneration frequency was 33.3% when stolon tissues with a node were cultured on the regeneration medium supplemented with 0.5 mg/L 2,4-D and 1 mg/L of kinetin. Comparing the basal media tested, MS medium showed higher plant regeneration performance than N6 or SH medium. Addition of 5 mg/L $AgNO_3$ with 10 mg/L cysteine improved frequency of plant regeneration up to 40%. Among different carbon sources, 3% sucrose was found to show the best for regeneration frequency. This rapid and efficient plant regeneration system would be useful for using genetic transformation experiments of manilagrass without intervening callus-mediated regeneration.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.29-34
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• 1994

The possibility that DNA could move out of the single cells isolated from geranium (Pelargonium zonale hybrids, 'PintoScarlet') callus was determined by the elechophoretic DNA analysis after treatment of low pH, various concentrations of KNO$_3$, 2,4-D, and GA$_3$ followed by the centrifugal force, all of which are hewn to and the physico-chemical properties of the cell wall. The centrifugal force of l,800 xg was need for DNA migration after the above treatment, but 7k300 xg was required without the treatments. In this experiment the optimum concentration (300 mg/L) of sodium dodecyl sulfate (SDS) used as an anion detergent to collect the negatively charged DNA was very critical not to damage the cell wall It can be concluded that the centrifugal force played a key role for the DNA migration through the cell wall, and the treatments of low pH (4.0), 0.5% KNO$_3$, 1.5 mg/L GA$_3$and 1mg/L 2,4-D further increased the DNA migration.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.95-99
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• 1995

We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.23-28
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• 1994

This study was rimed out to examine the difference in the cell vitality between mesophyII protoplast (MP) and paraveinal mesophyII protoplast (PVMP) of Nicotiana tabaccum 'Xanti', Petunia hybrida 'Blue Star' and Chrysanthemum morifolium 'Baeckwang' by using urea permeability technique. The effects of various enzyme solutions and incubation time, NAA and thidiazron on plant regeneration from isolated protoplasts were also investigated. The vibratome technique was used for protoplast isolation and urea permeability test because the fresh living, thin tissue stripes (50 ${\mu}{\textrm}{m}$ of thickness) could be obtained with minimal damage with the vibratome. For the three plants examined, the urea permeability on the tested tissue stripes was relatively higher in PVMP than in MP by about Ks = 2.0 $\times$ 10$^{-5}$ cm/sec. The treatment of an enzyme mixture of 1.5% cellulase R-10, 1% Driselase, 0.5% Macerozyme R-10, and 0.5% Pectinase for 4 to 8 h was effective on the isolation of PVMP. The highest frequency of callus formation and plant regeneration from the isolated protoplasts was obtained with NAA 2 mg/L and thidiazuron 0.01 mg/L. Furthermore, the results demonstrated that cell devision and plantlet regeneration was more frequent in the PVMP than in the MP of the same leaf or plant We, therefore, conclude that UM is an excellent experimental material for the callus formation and regeneration from isolated protoplasts.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.140-145
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• 2014

This study were carried out for selection of proper transformation variety and development of efficient regeneration and transformation methods. The number of shoot in commercial varieties of gourd plant were 0 ~ 7.3. and fusarium wilt resistant pure lines were 2.0 ~ 6.5 per dish containing on MS medium supplemented with 3 mg/L BA. The shoot regeneration frequency of fusarium wilt resistant pure lines were wide variation on the deviation. The expression of GFP was high 67% and 100% at the co-cultivation with Agrobacterium. The effective shoot regeneration plant hormone were combination BA and 2,4-D. The number and elongation condition of shoot was good after 4 weeks change with MS medium supplemented with 1 mg/L BA. Effective callus production plant hormone were combination of 3 mg/L BA and 0.1 mg/L 2.4-D.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.134-139
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• 2014

The aim of this study was to establish the condition of regeneration for white balloon flower (Platycodon grandiflorum DC. cv. Jangback) and to manage for the raising of seedling with in vitro regenerated plants. It was examined that 0.5 mg/L of NAA and 1.0 mg/L of BA was the best composition for the callus and shoot induction (up to 600%). NAA was better than IBA for the induction of root and it took 16.9 days for the induction of rooting on the MS soild media containing 0.5 mg/L of NAA and the final rooting ratio was up to 75%. Out of 5 different bed soils purchased from local market, "Tosil" was identified to be the best for the acclimation and growth of in vitro regenerated balloon flower. In detail, on 8 weeks after planting of in vitro regenerated plants in pots containing "Tosil" bed soils, the plant hight was increased up to 2-fold (12.8 cm), 3.5-fold (27) for the number of leaf and 1.5-fold (4.5 cm) for the leaf length when compared to the other four bed soils, respectively. Our preliminary results indicate that it is possible to prevent the occurrence of blue balloon flower in the massive cultivated area of white balloon flower by providing the seedlings raised from in vitro regenerated plants.

• Journal of Korean Society of Forest Science
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• v.95 no.4
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• pp.393-397
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• 2006

Thuja koraiensis Nak. is a short and creeping evergreen shrub which reaches about 3 m in height and only occurs in the northeast China and in high mountains over the Korea. It's designated as a rare and endangered tree species in Korea and DD (data deficient) in Red List Category & Criteria of IUCN. This study was carried out to develop the propagation technique by cutting for conservation of genetic resources of T. koraiensis. The rooting responses of branch cuttings, obtained from hard (May) and semi-hard wood shots (August) to three plant growth regulators (PGRs), namely, IAA, IBA, and NAA applied at various concentrations (0, 100, 500, 1000, 2000, and 3000 mg/l) were examined in sand and mixed soil media. Percentage of rooting showed significant difference between cutting time, among kinds and among concentration of PGRs. The optimum cutting time was April to May in hardwood cutting. The application of IAA 1000 mg/l and NAA 500mg/l were effective in callus formation and rooting of cutting. Relatively, rooting of cutting of the control taken in May was above 93%.

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.259-264
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• 2002

Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n＋4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.123-127
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• 2002

Effect of cytokinins (BA, TDZ, zeatin, 2iP, and kinetin) applied either singly or in combination on in vitro growth of two grape cultivars ('Cabernet Sauvignon' and 'Campbell Early') was investigated as a serial work for mass production of grapevine nursery stocks. In single treatment, shoot growth of two cultivars was most favorable in control. Shoot proliferation was satisfactory with 10 $\mu$M BA regardless of cultivars and cytokinin combinations, followed by TDZ. Other treatments resulted in very poor or no branching. Total explants ready for subculture produced by 10 $\mu$M BA outnumbered those by other treatments. TDZ was also effective. TDZ significantly increased the fresh weight and callus formation while shoot growth was unsatisfactory. Shoot growth response of two cultivars in combined treatments was also most favorable in control as was in single treatments. When TDZ was combined with zeatin, 2iP, and kinetin which failed to induce branching, proliferous branching was induced though the shoot number was behind that of single treatments of BA and TDZ. TDZ was very effective for total number of explants and fresh weight, showing 10-fold increase.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.129-134
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• 2002

194 plantlets regenerated from leaf explants of boxthorn 'Cheonyang' were cultivated to investigate their morphological characteristics in the field for 2 years. Based on the morphology of leaves, 66.1% of them had elliptical type leaf, the same as that of mother plants, while 22.2% in oval type, 7.2% in obovate type, 2.6% in long-obvate type and 2.1% in lanceolate type. They were classified to 4 groups; group A was selected with both high fruit size and fruit yield, group B with only high fruit size, group C with larger or thicken leaf, and group D with multiple brenches. In comparision of production efficiency between the selected groups and mother plants, group A (99741, 99781, 99854, 99870 and 99886) were longer (2.1 to 2.7 mm) in length of fruits and higher in fruit production (15 to 30%) as compared to mother plants. Croup D (99797 and 99892) was higher in leaf production (7.2%) as compared to boxthorn 'CL1-48', which is the highest in leaf production among boxthorn veriaties.