• Korean Journal of Plant Resources
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• v.22 no.4
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• pp.337-342
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• 2009

Present studies were conducted to evaluate the effects of medium strength(MS and Hyponex), carbon sources and their concentrations, agar concentrations, and inoculation amounts on prothallus propagation of Pterdium aquilinum var. latiusculum(Desv.) Underw. ex Hell in vitro. The optimum MS medium strength for prothallus propagation was 2MS concentration. Phosphate source was most effective for prothallus growth of P. aquilinum var. latisculum. The addition of 1% sucrose or glucose to MS medium promoted prothallus multiplication. Growth of prothallus was not affected by agar concentration. Propagation of homogenized prothallus was vigorous even in liquid medium. Chopped gametophytes(100 and 200 mg) were inoculated on 250 ml ${\Delta}$flask with 100 mL of 2MS concentration medium and suspension culture was done at 100 rpm for 22 days. After 20 days, prothallus multiplication slowed down, so 100 mg of chopped prothalli is recommended for initial inoculation, since initial amount of inoculum did not affect subsequent prothallus multiplication. Consequently after 20 days of suspension culture, prothallus should be subcultured or transplanted outside of growing vessels.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.69-76
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• 2011

This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing $0.4\;mg\;l^{-1}$ thiamine HCl, $100\;mg\;l^{-1}$ myo-inositol, $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at $25^{\circ}C$, the plating efficiency of cabbage protoplasts reached to $22.5{\pm}2.9%$ when cultured in modified MS medium supplemented with $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and 8% (w/v) of myo-inositol at a density of $2{\times}10^5$ protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing $2\;mgl^{-1}$ BA and $0.5\;mgl^{-1}$ NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.111-118
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• 2009

Multiple shoot induction and plant regeneration using apical bud and nodal explants of 100 year old tree of Anthocephalus cadamba, an important sacred and medicinal tree in India was achieved for the first time. Aseptic explants cultured in Murashige and Skoog (MS) medium augmented with different concentrations of BAP (0.1, 0.5, 1.0, 2.5, 5.0 and 10 mg/l), when maintained for 60 days, healthy shoots were induced in presence of BAP (1 mg/l). Lower concentrations of BAP (0.1 - 0.5 mg/l) induced only one shoot per explant. Increase in number of shoots per explant was observed in presence of higher concentrations of BAP (2.5, 5.0 and 10 mg/l). However, elongation of shoots was completely inhibited. Bud break and shoot regeneration was largely associated with seasonal factors. Apical buds cultured during June to August exhibited early bud break within two weeks of initial culture. In rest of the months, bud break and shoot regeneration was very slow irrespective of the various concentrations of BAP used in the medium. Explants sourced from three different maturity levels of shoots indicated that actively growing shoots from the mother plant with 1 - 2 nodal segments was more suitable for culture initiation than the explants collected from mature shoots at dormant stage. Regenerated shoots with 2 - 3 pairs of leaves when transferred to half strength MS medium fortified with IBA (1 mg/l), 60% of the shoots induced healthy roots, indicating the possibility of large scale micropropagation.

• Current Research on Agriculture and Life Sciences
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• v.29
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• pp.21-27
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• 2011

This study was carried out to produce multiple shoots and bulblets from flower stalk tissue cultures of Muscari armeniacum LEICHTLIN ex BAKER 'Early Giant', which were cultured in the half strength Murashige & Skoog's (MS) medium supplemented with auxin, NAA in combination with kinetin, BA, and TDZ, alone and/or. In flower stalk tissue culture, upper part explant was the most suitable as a source of culture material. Direct shoot formation was much more favorable in half strength MS medium supplemented with $0.1mg{\cdot}L^{-1}$ NAA and $1.0mg{\cdot}L^{-1}$ TDZ. On the other hand, bulblet formation was increased when cultured in half strength MS medium added with $0.01mg{\cdot}L^{-1}$ NAA and $0.1mg{\cdot}L^{-1}$, $0.2mg{\cdot}L^{-1}$ kinetin. Acclimatized plant flowered during the second year of the growing period without any phenotypic variations and formed average 1.5 bulblets per mother bulb.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.218-223
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• 1993

Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

• International Journal of Oral Biology
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• v.35 no.1
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• pp.1-5
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• 2010

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells ($1.09\;{\times}\;10^5\;cells/ml$). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% ($3.0\;{\times}\;10^4\;cells/ml$) or 20% KSR ($4.8\;{\times}\;10^4\;cells/ml$) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

• 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.

• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.4
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• pp.353-360
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• 2012

This study was conducted to evaluate the nutritive value and the quality of ensiled kenaf after fermentation with three cultivars at three different times of harvesting. Experimental plot were allocated with three harvest date (Early;8/3, medium;8/15 and late;8/28) and three cultivars (Tainung-a, Everglade, Whitten). The DM (dry matter) yield increased with maturity in all three cultivars, especially in Whitten which showed the highest yield at each harvest time. The DM content in Whitten at late harvesting time was higher than other treatments (231 g $kg^{-1}$ DM). The CP (crude protein) contents of the kenaf silage of all three cultivars ranged from 151 to 164 g $kg^{-1}$. Highest content of NDF (neutral detergent fiber) was observed in Everglade at medium harvesting date, but the ADF (acid detergent fiber) content was highest in Everglade at early harvesting date (p<0.05). All treatments produced a pH less than 4.0, which is sufficient for stable storage. The pH of Tainung-a was higher than other cultivars at all harvesting times (p<0.05). Whitten showed the higher content of lactic acid (25.8 g $kg^{-1}$ DM) showed at early harvesting date than other cultivars (p<0.05), while the acetic acid content (21.1 g $kg^{-1}$ DM) was higher in Tainung-a at medium harvesting date (p<0.05). No significant difference was observed in ammonia-Nand butyric acid concentrations among all treatments. These results indicate that a kenaf silage could be used as fodder for ruminants. Especially, the Whitten harvested at late growing stage showed promise as a forage silage crop under Koran environments.

• Journal of Information Technology Applications and Management
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• v.24 no.2
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• pp.25-37
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• 2017

The aim of this study is to analyze the effect of the growing activities of large purchasing enterprises on the trust of small and medium size contractors, which is mediated by 'purchase accompany activity'. This subject study includes the first and second contractor companies in Korea. The results of this study are as follow. First, the knowledge and cost price sharing activities of major purchase vendor with cooperative company had a positive effect on the trust of cooperative company, however this was not so with the information sharing activities in that case. Second, the trust of contractors on large purchasing enterprises was positively influenced by sharing activities with purchasing activities of major company. Third, the knowledge and cost price sharing activities of purchasing enterprise with contractors on the sharing activities were mediated by the trust of contracting company on the large purchase enterprise. According to this study, the mediated effect of trust relationship can influence a contracting company. Whereas preceding studies of sharing activities were conducted about unidirectional instruct of large purchase enterprises to small and medium size contracting companies, this study was conducted on bidirectional effects, which included knowledge, cost price, information sharing activities between contracting companies and large purchasing enterprises. Conclusively, this study showed the possibility of spreading cognitive and change about mutual benefits strategy between small and medium size contracting company and large purchasing enterprise.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.189-195
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• 2001

This study was carried out to investigate the effects of in vitro culture condition and among lines on growth pattern of lateral buds from nodal cuttings of Phalaenopsis flower stalks. The ratio of bud growing into shoot from nodal cuttings of flower stalks were 90.9% and 54.4% on MS and hyponex medium, respectively. The number of buds grown vegetatively were increased remarkably on the MS medium containing 5 mg/L BA. The rate of buds grown vegetatively was higher in basal and middle parts than in upper part of flower stalks. The flower stalk sections cultured at 25~28$^{\circ}C$ showed the highest ratio of vegetative growth. Medium contamination was decreased by pretreatment of etiolation to the flower stalk. However, the pretreatment did not show specific effects on shoot development and reduction of phenolic compound. Average shoot number which was formed from flower stalk segments in 27 of 30 accessions were 3.17, while high number of shoots were obtained from Phal. 3020 and Phal. 3039. The growth pattern of lateral buds in F$_1$hybrids was similar to that of their parents.