• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.157-163
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• 2004

An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.4
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• pp.322-328
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• 2006

A series of methanol extracts from leaf and root parts in spring- and winter-barley plants were assayed to determine their allelopathy and antioxidant activity. The methanol extracts applied on filter paper in a Petri-dish significantly inhibited root growth of Chinese milkvetch (Astragalus sinicus L.) seedlings. Leaf extracts at 25 and 50 g $L^{-1}$ inhibited root growth of Chinese milkvetch seedlings more than root extracts. No difference in phytotoxic effects of spring- and winter-barley seedlings extracts on root growth of Chinese milkvetch was observed. Methanol extracts dose-dependently increased DPPH free radical scavenging activity in vitro. DPPH free radical scavenging activity was higher in the methanol extracts from winter-barley seedlings than in those from spring-barley seedlings, and from leaf extracts than from root extracts. The antioxidant potential of the individual fraction from the methanol extracts of spring-barley seedlings was in order of n-butanol>ethyl acetate>water>chloroform>n-hexane fraction. By means of HPLC analysis, spring-barley (200.62 mg $100g^{-1}$) had more amount of total phenol acid than winter-barley (114.08 mg $100g^{-1}$). Especially, ferulic acid was detected in spring-barley extract (183.46 mg $100g^{-1}$) as the greatest amount. These results suggest that early seedlings of barley plants had potent allelopathy and antioxidant activity, and their activities were differently exhibited depending on plant parts and growing condition.

• KSBB Journal
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• v.11 no.3
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• pp.263-269
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• 1996

Anticancer agent taxol was synthesized from baccatin III which was extracted from cell cultures of European yew, Taxus baccata Pendula. Callus and suspension cultures of T. baccata Pendula showed the formation baccatin III. The content of baccatin III in cell cultures reached 0.015% (w/w) on dry weight basis. The semisynthetic approaches were made with baccatin III and taxol side chain. To prepare taxol side chain, (-)-N-((S)-2-hydroxy-1-phenylethyl) benzamide was synthesized first from (S)-(+)-phenylglycine. Then (-)-N-((IS,2S)-2-hydroxy-1-phenyl-3-butenyl) benzamide was synthesized with vinyl magnesium bromide. The synthesis of (2R, 3S)-(-)-2-(1-ethoxyethoxy)-3-phenyl-3-(phenylmethanamido) propanoic acid with RuCl3, catalyst was the final step to prepare taxol side chain. The semisynthetic reactions yielded 0.0002% taxol, 0.0005% 7-epi-10-deacetyltaxol, and unidentified taxanes on dry weight basis. It is suggested that the semisynthesis of taxol from baccatin III could be an alternative source of taxol and related taxanes.

• Plant Biotechnology Reports
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• v.2 no.4
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• pp.245-252
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• 2008

The genetic status of somatic embryo-derived plantlets of Cymbopogon flexuosus was examined by randomly amplified polymorphic DNA (RAPD) analysis. Auxins such as 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1-4 mg/l) were used in Murashige and Skoog (MS) medium for induction of calli from rhizomatous explants of Cymbopogon flexuosus. Optimum calli were induced on MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (2, 4-D) (3.5 mg/l) alone or in combination with $N^6-benzyladenine$ (2 mg/l). Somatic embryogenesis was achieved from long term calli when cultured on MS medium containing 2, 4-dichlorophenoxyacetic acid (2, 4-D) (2 mg/l) along with $N^6-benzyladenine$ (BA) (1-2 mg/l). Regeneration was achieved when freshly induced embryogenic calli were sub-cultured on MS medium supplemented with $N^6-benzyladenine$ (3 mg/l) alone. Long-term cultured embryos showed profuse minute rooting on regeneration medium supplemented with N6 -benzyladenine (3 mg/l). Microshoots were rooted in the presence of indole-butyric acid (IBA) (2 mg/l). DNA samples from the mother plant and 18 randomly selected regenerated plants from a single callus were subjected to RAPD analysis with 6 arbitrary decamer primers for the selection of putative somaclones. A total of 64 band positions were scored, out of which 19 RAPD bands were polymorphic. From genetic similarity coefficient based on RAPD band data sharing, it was found that the majority of the clones were almost identical or more than 92% similar to the mother plant, except CL2 and CL9 (66%) which showed highest degree of genetic change with CL2 and CL9 showing presence of two non-parental bands each.

• FLOWER RESEARCH JOURNAL
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• v.17 no.1
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• pp.23-28
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• 2009

This study was conducted to examine the effect of plant growth regulators on the formation of adventitious bud callus(ABC) and plant regeneration in shoot tip culture of Zantedeschia spp. 'Florex Gold'. Treatment of $0.1mgL^{-1}\;N$-phenyl-N'-1,2,3-thiadiazol-5-ylurea(thidiazuron, TDZ) was more promotive for formation of ABC in shoot tip culture than 6-benzylaminopurine(BA) treatment, and short shoots were developed. Comparing to treatment of BA, mixing treatment of BA and 1-naphthaleneacetic acid(NAA) inhibited the formation of ABC and multiple shoot. The proliferation of ABC derived from sections(0.3 cm) of ABC produced by shoot tip culture in medium supplemented with $0.1mgL^{-1}\;TDZ$ was more effective in medium with $0.1mgL^{-1}\;TDZ$ or $2.0mgL^{-1}\;BA$ than the other treatments. The shoot regeneration and growth from sections of ABC was more promotive in treatment of $0.001mgL^{-1}\;TDZ$. Also, the root growth from sections of ABC was better in medium with $0.001mgL^{-1}\;TDZ$ or $0.2mgL^{-1}\;BA$. Consequently, in vitro mass production of Zantedeschia spp. 'Florex Gold' can be obtained via indirect organogenesis through plant regeneration and proliferation of ABC which was derived from shoot tip culture at $0.1mgL^{-1}\;TDZ$.

• Journal of Plant Biotechnology
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• v.7 no.2
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• pp.97-103
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• 2005

This study was carried out to obtain basic information for gene manipulation in potent edible tobacco (Nicotiana tabacum cv. TI 516). N. tabacum cv. TI 516 is a plant for a possible candidate to use as an edible vaccine, since it contains a low level of nicotine. The effective plant regeneration system through leaf disc culture was achieved using a MS basal medium supplemented with 0.1 mg $1^{-1}$ NAA and 0.5 mg $1^{-1}$ BA. In order to transform the N. tabacum cv. TI 516 with the green fluorescent protein (GFP) gene, Agrobacterium tumefaciens LBA 4404 containing the GFP gene was used. Genomic PCR confirmed the integration of the GFP gene into nuclear genome of transgenic plants. Expression of the GFP gene was identified in callus, apical meristem and root tissue of transgenic N. tabacum cv. TI 516 plants using fluorescence microscopy. Western blot analysis revealed the expression of GFP protein in the transgenic edible tobacco plants. The amount of GFP protein detected in the transgenic tobacco plants was approximately 0.16% of the total soluble plant protein (TSP), which was determined by ELISA.

• Archives of Plastic Surgery
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• v.34 no.3
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• pp.298-304
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• 2007

Purpose: This study was undertaken to investigate the osteogenic induction potential of PGA & FBS mixture on a calvarial defect in the rabbit. Methods: Twenty New zealand white rabbit, weighing from 3.5-4kg were allocated into each of the three groups. Four 8 mm sized bone defects were made on the parietal bone by drilling. In group I, the bony defects were implanted with $50{\mu}m$ thickness film containing mixture of PGA and FBS. In group II, with PGA only film, & in group III, the bony defects were left with no implants. Results were evaluated by using morphologic change, radiographic study, biochemical study and histologic examination at 1 week (group I n=7, group II n=7, group III n=14), 2 weeks (group I n=6, group II n=6, group III n=12) and 3 weeks (group I n=7, group II n=7, group III n=14) following implantation. Results: In the morphologic & radiographic study, the formation and corticalization of callus were observed earlier in group I than in groups II and III (p < 0.05). In histological examination, group I showed more abundant and faster new bone formation than in group II and III. In biochemical analysis, group I displayed more activity than in group II and III. Group I also showed more abundant osteopontin, osteocalcin than groups II and III. Conclusion: In conclusion, the results demonstrate that the mixture of PGA and FBS has an effect on osteoblastic formation in the rabbit model. It is considered that further evaluation of long term results on resorption, immunologic tissue reaction and response of applied mixture in the human model will be needed.

• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.69-74
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• 2012

Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.33 no.3
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• pp.248-253
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• 1988

The longer pollen stage grew to flowering stage, the higher anther respiratory rate in vivo became. and it was rapidly increased just before flowering. The anther respiratory rate in vitro showed the first and second peak points after 3-7 days and 9-1l days in culture, respectively, and fastest and highest in Daecheongbyeo with high sporophytic potentiality. It was lower in cold-pretreatment than non-treatment at the early days, but higher from 15 days after culture. The frequency of browning anthers was promoted by cold-pretreatment. The respiratory rate was not different between uncolored and browned anthers at 12 days, but it was higher in browned anthers after 24 days in culture.

• Journal of Ginseng Research
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• v.36 no.3
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• pp.322-326
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• 2012

Results of karyological study of intact plants and some callus lines of Panax ginseng are presented. In the native plants of P. ginseng the nucleus with 1 nucleolus (90%) dominate, and nucleus with 2 nucleoli is rare. One nucleolar nucleus also dominate in interphase nuclei of cells of cultivated P. ginseng (from 2006), but we also found nucleus with 2 to 3 nucleoli in the same cell lines. Interphase nuclei of P. ginseng in long cultivated lines (from 1988) contain 1 to 9 nucleoli, with a predominance of nuclei containing from 3 to 4 nucleoli. It was shown that long-time cells (cultivated since 1988) had cytogenetic changes such as increase level of polyploid and aneuploid cells, increase of nucleoli number into interphase nucleus and decrease of nuclei/nucleoli ratio. These long-time cultivated cells had very low ginsenoside content.