• Journal of Life Science
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• v.11 no.3
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• pp.254-258
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• 2001

The biochemical change during regeneration of perilla callus were investigated by comparing total protein and lipid contents, protein band pattern in SDS-PAGE, and fatty acid composition in the calli cultured for various period(0, 1, 3, 5 and 6 weeks) Calli were induced from cotyledon and hypocotyl explants of peplants of perilla on perilla on MS medium containing BA(0.5 mg/L) and NAA(0.5mg/L). The protein contents reached the peak at 3 weeks after induction of calli, and then was decreased. Total lipid contents was decreased as the culture period increased. The band pattern of polypeptides showed that 30KD and 45KD polypeptides and 22KD and 45KD polypetides were major proteins in the cotyledon and hypocotyl explants, respectively. However increase of culture period only 30KD protein was highly accumulated.

• Journal of Ginseng Research
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• v.6 no.2
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• pp.162-167
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• 1982

Calli and leaflets of ginseng (Panax ginseng C.A. Meyer) were cultured on 1/2MS media supplement. with kinetin, 2 iP, NAA, 2,4-D and IBA to assess their capacity to regenerate embryoids and organs. Root calli produced numerous embryoids and shoots in 1/2MS medium supplemented with 2 mg/l NAA and 2mg/s 2iP, and the combination of 2 iP and NAA was more effective than the combination of kinetin and NAA in induction of embryoid and shoot from root calli. Culture of leaflet in the medium supplemented with IBA resulted in profuse root regeneration.

• Proceedings of the Ginseng society Conference
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• 1993.09a
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• pp.143-149
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• 1993

Ginseng root explants and calli induced from selected cell lines were cultured on modified Murashige and Skoog's media supplemented with different concentrations of organic or inorganic compounds and plant growth requlators to clarify the effects of chemical composition and plant growth regulators in the medium on the growth of ginseng calli and the production of ginseng saponin. For optimum growth of calli, the concentrations of 2, 4-D and sucrose were the range of 1 to 3 mg/${\ell}$l and 1 to $3\%,$ respectively. And it was clarified that sucrose, nitrogen, phosphate, calcium, magmesian plant growth regulators and their concentrations influcenced the relative biosynthesis of saponin in tissue cultures of Panax ginseng. The patterns of ginsenosides, pharmacologically useful component, were different among the cell lines and contents of ginsenosides were much higher in selected cell lines than in original cell line.

• Journal of Korean Society of Forest Science
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• v.86 no.3
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• pp.261-269
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• 1997

Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and ${\beta}$-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced about $25.75{\pm}2.77$(per ovule), $11.43{\pm}1.22$(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.73-80
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• 2003

Useful Dianthus species were collected and selected from two native and seven foreign species distributed in Gangwon province. For in vitro breeding,. callus was induced from the explants of apical meristem, leaf, stem and the in vitro adventitious shoots on MS basal medium with 2.0 mg/L 2,4-D and 0.5 mg/L BA at 27$^{\circ}C$ under continuous light. After 3 weeks of culture, calli initiated the most highly from the leaf explants of D. chinensis Organogenic calli were able to be selected from the adventitious shoot-derived calli. For shoot regeneration, these organogenic calli were cultured on MS medium with the combination of 0.1 mg/L NAA＋1.0 mg/L BA under continuous light. Multiple shoots were proliferated with low frequency (about 30%) from those adventitious shootderived calli. Also, shoots initiated directly from the adventitious shoot explants without callus formation at high frequency of 52% when cultured on N6 medium containing 0.1 mg/L NAA and 1.0 mg/L BA in D. gratianopol. Multiple shoots and plantlets grew well and rooted on MS medium supplemented with 0.1 mg/L NAA. Regenerants with well-developed roots were transferred to 8-cm pots containing vermiculite at 85% relative humidity and 27$^{\circ}C$ These plantlets were acclimatized in artificial soil mixture and transferred to the greenhouse for flowering with normal phenotypes. M28 Mutant line was selected with white flowers from 0.03M EMS-treated organogenic calli derived from in vitro adventitious shoot explants of D. chinensis and set seeds.

• Journal of Plant Biology
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• v.32 no.4
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• pp.313-322
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• 1989

A system was established for induction of multi-shoots and plant regeneration from mesophyll protoplasts of alfalfa, Medicago sativa L. cv. Vernal. Different hormonal effects were tested at each step of protoplast culture, i.e. cell division in modified Kao's liquid medium (K566-7). calli formation on SH semi solid medium, and multi-shoot regeneration from calli on SHa and SHb solid media. Frequency of multi-shoots and plant regeneration was affected by various combinations of phytohormones in final step. The evaluation of multi-shoots induction systems via protoplast culture was discused.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1390-1394
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• 2004

We have achieved efficient transformation system for forage-type tall fescue plants by Agrobacterium tumefaciens. Mature seed-derived embryogenic calli were infected and co-cultivated with each of three A. tumefaciens strains, all of which harbored a standard binary vector pIG121Hm encoding the neomycin phosphotransferase II (NPTII), hygromycin phosphotransferase (HPT) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. Transformation efficiency was influenced by the A. tumefaciens strain, addition of the phenolic compound acetosyringone and duration of vacuum treatment. Of the three A. tumefaciens strains tested, EHA101/pIG121Hm was found to be most effective followed by GV3101/pIG121Hm and LBA4404/pIG121Hm for transient GUS expression after 3 days co-cultivation. Inclusion of 100 $\mu$M acetosyringone in both the inoculation and co-cultivation media lead to an improvement in transient GUS expression observed in targeted calli. Vacuum treatment during infection of calli with A. tumefaciens strains increased transformation efficiency. The highest stable transformation efficiency of transgenic plants was obtained when mature seed-derived calli infected with A. tumefaciens EHA101/pIG121Hm in the presence of 100 $\mu$M acetosyringone and vacuum treatment for 30 min. Southern blot analysis indicated integration of the transgene into the genome of tall fescue. The transformation system developed in this study would be useful for Agrobacterium-mediated genetic transformation of tall fescue plants with genes of agronomic importance.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.283-290
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• 1995

This experiment was canted out to obtain embryogenic callus and to understand developmental mechanism of somatic embryogenesis in Oenanthe javanica ($B_{L.}$) DC. experiments included the examination of explant source and media for embryogenic callus production and the observation of developmental pattern of embryogenic cells and non-embryogenic cells. Embryogenic calli were formed on zygotic pro-embryos together with their endosperms when they were cultured on Ms media containing 1.0mg/L 2,4-D. Embryogenic calli were also formed on the intact surface in vitro grown stem or petiole segmentsafrer 6-8 weeks of culture, whereas non-embryogenic calli were formed on cut surfaces of the stem and petiole after 2 weeks of culture. Non-embryogenic calli were rhizogenic in suspension and solid media culture.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.233-238
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• 2003

Sweet potato cells derived from Yulmi were isolated from embryogenic callus and irradiated with 50 Gy dose. Resistant cells were selected on a MS medium containing 1.0 mM S-aminoethyl-L-cysteine (AEC). This level of AEC approximately inhibits non-selected wild type cells. The callus resistant to this analog of lysine was subcultured for 30 days in absence of AEC to proliferate. The three resistant calli (AR-1, AR-2 and AR-3) with better growth were divvied into 0.5~1mm diameter and placed on MS medium with 0, 0.4, 0.6, 0.8 and 1.0 mM AEC. There are considerable growth difference between control callus and AEC resistant callus on the AEC-medium. The selected calli were placed on the hormone-free medium for regeneration. Three plantlets, five plantlets and six plantlets were recovered from AR-1, AR-2 and AR-3 calli, respectively. Each two regenerants in AR-1, AR-2 and AR3 were randomly selected for RAPD and SDS PAGE analysis. RAPD polymorph isms between Yulmi and AEC resistant plant from irradiated calli were detected in several Wako primers. Also, it was identified that two AEC resistant plants had higher protein than the original variety Yulmi.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.233-239
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• 2003

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.