• Plant Biotechnology Reports
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• v.2 no.3
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• pp.171-177
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• 2008

This paper reports an improved protocol for isolation, culture and regeneration of Lotus corniculatus protoplasts. A range of parameters which influence the isolation of L. corniculatus protoplasts were investigated, i.e., enzyme combination, tissue type, incubation period and osmolarity level. Of three enzyme combinations tested, the highest yield of viable protoplasts was achieved with the combination of 2% Cellulase Onozuka RS, 1% Macerozyme R-10, 0.5% Driselase and 0.2% Pectolyase. The use of etiolated cotyledon tissue as a source for protoplast isolation proved vital in obtaining substantially higher protoplast yields than previously reported. Culture of the protoplasts on a nitrocellulose membrane with a Lolium perenne feeder-layer on the sequential series of PEL medium was highly successful in the formation of microcolonies with plating efficiencies 3-10 times greater than previous studies. Shoot regeneration and intact plants were achieved from 46% of protoplast-derived cell colonies.

• Journal of the Korean Professional Engineers Association
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• v.18 no.2
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• pp.16-22
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• 1985

This study was carried out to evaluate the quality of soybean sprouts grown under three different sources of light. The soybean was germinated under light through blue (251 ux), red (751 ux) and white (50 to 200 lux) polyethylene films at 25$^{\circ}C$ for 10hr./day. Vitamin C, chlorophyll, cellulose and total protein contents were determined and texture was evaluated by tasting soybean sprouts soup. protein pattern in the soybean sprouts were investigated using polyacrylamide gel electrophoresis. Vitamin C, chlorophyll and cellulose contents were increased by white light intensity. The texture of the sprouts grown under white light (50 lux, 100 lux) was better than darkness, but fought under 200 lux. Vitamin C contents of soybean sprouts grown under various sources of light (in order of light : blue > white > red) were higher than theses of ones grown in the darkness. Biosynthesis of chlorophyll was not correlated to Vitamin C content. Total protein contents of cotyledon was not changed significantly under light irradiation. But the soybean sprouts grown under different quality of light, hypocotyl was higher than those grown darkness. (37% and 20% higher for blue light and white light) Densitometric tracing of disc polyacrylamide gel electrophoretic patterns showed that protein of hypocotyl under white light had more high molecular weight protein.

• Journal of Bio-Environment Control
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• v.6 no.4
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• pp.317-323
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• 1997

Incidence of soybean sprout rot by Erwinia carotovora subsp. carotovora was examined under several artificial conditions. Under higher temperatures over 3$0^{\circ}C$, disease incidence was increased and the rate of soft rot incidence was 22% at 35$^{\circ}C$. Artificial injuries of inner cotyledon and seed coat induced the disease above 70% and inhibited the soybean sprout growth. Relative humidity above 90% increased the soft rot to 33% and inhibited soybean sprout growth. When the leaked water collected from soybean sprout was reused for irrigation, the disease incidence was increased.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.245-250
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• 2003

An efficient procedure was developed for adventitious shoot bud induction and plantlet regeneration from various explants of the ten genotypes of Pepper (Capsicum annuum L.) using Thidiazuron (TDZ). Among various treatments at 1.0-3.0 mg/L TDZ Induced maximum number of adventitious shoots depending upon the explant type and genotype compared to other treatments. Among the explants tested, leaf induced maximum number of adventitious shoots than the cotyledons. TDZ-mediated organo-genesis was possible in 10 pepper cultivars, the extent of the response being genotype-dependent. Of the ten genotypes tested, C. annuum cvs CA960, $G_4$ and X-235 were produced maximum number of adventitious shoots and Sell was the least, and all other genotypes gave moderate response. Elongation of multiple shoots was observed on medium supplemented with SA (0.05 mg/L) in combination of IAA (0.05 mg/L). Differences in ability for in vitro shoot regeneration and elongation depend upon the variety and explant type. The elongated shoots were success. Fully rooted on MS medium containing at 1.0 mG/L IAA. Plantlets regenerated from different explants of ten genotypes were found to be diploid (2n=24) and were devoid of any chromosomal aberrations. Regenerated plants were successfully established in soil where 85-90% of them developed into morphologically normal and fertile plants.

• Korean Journal of Plant Tissue Culture
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• v.28 no.6
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• pp.353-357
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• 2001

Transformation of ginseng plants was achieved by biolistic system with cotyledon explants and callus using phosphinothricin acetyl-transferase (PAT) gene resisting to a herbicide of Bialaphos. The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 355 promoter. The introduced NPT II and PAT genes of the transgenic ginseng plants were successfully identified by the PCR, and the survival test on the medium with basta. The transgenic ginseng plants were propagated using repetitive secondary embryogenesis. The transgenic ginseng plantlets had normal structures of roots and shoots, and dormant buds for new year sprouting. We transferred the transgenic plants to greenhouse and observed the continuing growth until a new year.

• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.149-156
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• 2009

Application of exogenous polyamines (PAs) reduced the paraquat (PQ)-induced cotyledon injuries in radish seedling plants with 1 mM spermidine (Spd) being the most effective protectant. PQ injury symptoms in the cotyledons, e.g., large accumulation of $H_2O_2$, and losses of fresh weight, chlorophyll, and proteins, were significantly alleviated. Likewise, analysis of $H_2O_2$-scavenging enzymes such as catalase (CAT) and guaiacol peroxidase (GPX) showed that pretreatment with Spd among PAs remarkably increased total CAT activity and strongly retarded PQ-induced rapid decline in total GPX activity. In a native gel assay, one CAT isozyme (CAT1) and two GPX isozymes (GPX1 and a newly synthesized GPX isozyme) proved to be more responsible for PQ tolerance, as manifested by the strong increases in their activities by Spd pretreatment. Based on these results, we can suggest that PAs (especially 1 mM Spd) may function as antioxidant protectors by invoking CAT and GPX enzymes which control the endogenous $H_2O_2$ level in radish cotyledons exposed to PQ.

• Journal of Ginseng Research
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• v.16 no.1
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• pp.37-43
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• 1992

Structural changes of the endosperm of Panax ginseng C.A. Meyer from fertilization to germination were investigated by light microscope. The endosperm of the ginseng seed is cellular type. Since endosperm cells adjacent embryo continuously breakdown and disappear with the elongation of embryo, the real of endosperm is gradually decreased. As the anatropous ovules of immature seed with green seed coat developes more and more, ovary cells adjacent ovary cavity become abundant by the periclinal division, their size is decreased, hypotrophy of cell wall discern, and they are gradually differentiated in seed coat. Though embryo responds strongly to basic dye at the stage of completion of endosperm formation, tissue of endosperm responds to acidic dye positively Cell wall of embryo and endosperm are composed of primary cell wall not lignified. Endosperm cells adjacent embryo begin to breakdown in the endosperm tissue of indehiscent seed before the beginning of the after-ripening. Dehiscent seed of which seed coat is opened through after-ripening represent the form as a seedling in the result of embryo developments with the formation of organs; radicle, cotyledon, plumule. Umbilifom layer represents strong positive response to the toluidine blue and the basic function. Umbiliform layer that endosperm cells breakdown and disappear is observed clearly at the periphery of the embryo cotylemon, while slightly at the periphery of the radicle.

• The Plant Pathology Journal
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• v.19 no.4
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• pp.203-204
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• 2003

Decayed samples of marketed mungbean sprout were collected from Sacheon, Suncheon, and Dangjin cities in Korea. Initial symptom on hypocotyls was dark-brown diamond speck, which developed into enlarged sunken brownish-black spot with irregular margin, followed by softening yellowish decay. Brown speck on cotyledon further developed into irregular lesions. This study isolated the fungus Colletotrichum sp. The fungal colony was pale orange, which turned greenish gray after 1 week at $25^{\circ}C$. Colony of reverse side in a petri dish was pink. Neither conidiomata nor setae were present in the culture. Typical fusiform conidia sized 7.5-15.0$\times$2.5-2.9 $\mu\textrm$were hyaline, aseptate, smooth, and had salmon color in mass. Conidiogenous cells were phialidic, hyaline, smooth, and cylindrical with terminal distinct collarette. Basedon these mycological characteristics, the casual organism was identified as Colletotrichum acutatum. This is first report of Colletotrichum acutatum in Korea.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.55-58
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• 1994

Agrobacterium tumefaciens LABA4404 harboring plant binary vector, pBI121, was used for genetic transformation of lettuce (Lactuca sativa t.). Cotyledon segments were infected with A. tumefaciens LBA4404 by cocultivation method and regenerated. Regenerated letture was subject to molecular analyses for integration into plant nuclear genome and expression of ${\beta}$-glucumnidase (GUS) gene. Southern and Northern blot analyses demonstrated that GUS gene was integrated into plant nuclear genome and expressed into its mRNA. The expression of GUS gene into its protein was confirmed by specetrophotometric assay of GUS activity.