• Korean Journal of Plant Resources
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• v.19 no.3
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• pp.418-421
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• 2006

In this study, we surveyed the influence of IOA (iodoacetamide) and media upon protoplast fusion for the efficient production of the somatic hybrid among S. sisymbriifolium and other Solanum species (S. intergrifolium and S. toxicarium). Regardless of a breed, as the IOA concentration increases, the cell division tends to decrease. As the influence of the media on the colony formation, we could get 5 colonies from the fusion of S. sisymbriifolium and IOA-treated S. integrifolium protoplast, but none was observed in other fusions. As a result of analyzing their IDH isozyme, we found a somatic hybrid in 2 objects.

• Korean Journal of Microbiology
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• v.27 no.1
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• pp.27-34
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• 1989

In order to evaluate the applicability of enzyme electrophoresis for the identification of intra/interspecific hybride obtained by the protoplast fusion in Trichoderma, soluble proteins, intracellular soluble enzymes and extracellular $\beta$-glucosidase were analyzed by polyacrylamide gel electrophorsis. As the results, patterns of soluble protein, and isozyme patterns of peroxidase, malate dehydrogenase, and $\beta$-glucosidase in hydrids were defferent from those in parental and wild type strains. Therefore, it was established that the analysis of protein pattern by electrophoresis could be applied to isolate and identify the hybrids from the protoplast fusion.

• Archives of Pharmacal Research
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• v.16 no.4
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• pp.300-304
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• 1993

Genetic transformation of streptomyces gaespitosus by plasmid plJ 702 was camied out. Optimal conditions for the protoplast preparation of streptomyces casepitosus, its regeneration, and its transformation by plJ 702 were evaluated. Addition of 2% glycine to the culture broth was optimal for protoplast yield. Formation and regeneration of protoplasts were most efficient when the mycelium were harvested at between late log and stationary growth phase. The regeneration frequency of the protoplasts was 15% when the protoplats were regenerated on R2YE agar media containing 0.5M sucrose. Under the best condition for protoplats (M.W. 4,000) treatment for 2 minutes.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.274-277
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• 1990

The hyphal tip phenoloxidases of Coprinus congregatus were localized by the protoplast-concanavalin A method. Protoplast were generated from cultures grown on a solid medium which was solidified with a new gelling agent, Pluronid Polyol F127, instead of agar. the enzymes were associated with the cell membrane which might work as a transducer in the light recepter complex.

• Journal of Environmental Health Sciences
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• v.31 no.4 s.85
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• pp.327-331
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• 2005

Micromonosporas purpurea로부터 효율적 gentamicin 생산을 위해 protoplast fusion와 protoplast mutagenesis 방법이 검토 되었다. $CO^{60}\;irradiation\;(2.3{\times}10^5$ units, UV 3 min) 방법에 의해서 MP3-112, MP3-141, MP3-143을 분리 했다. 특히 MP3-143균주는 최대 gentamicin생산량이 얻어졌다. 개량된 MP3-143균주를 이용해서 탄소원 소비, 균체성장, 그리고 gentamicin 생산량이 batch culture에서 비교되었다. MP3-413와 parent 균주의 glucose 소비는 배양 2일과 3일 후에 각각 완전히 이루어졌다. 그러나 균체성장과 Soybean oil 소비는 비슷한 결과 얻어졌다. Gentamicin최대 생산량은 배양 5일 후 29756 U/ml였다. 이 결과는 parent 균주에 비해 생산량이 5.6배 증가했다.

• Biomolecules & Therapeutics
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• v.8 no.3
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• pp.235-240
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• 2000

Antitumor effect of LC43, a protein-bound ploysaccharide (M.W. 43 kDa) that was purified from intergeneric protoplast fusant of Lentinus edodes and Coriolus versicolor, was elucidated against mouse sarcoma 180 cell in vitro and in vivo. By injecting LC43 into ICR mice bearing solid or ascitic sarcoma 180, tumor regression and survival rates were investigated. To examine the effects of LC43 on immunopotentiation activity. immunoorgan weight, B cell differentiation, T cell activity and macrophage activation were determined. LC43 showed antitumor effects against both solid tumor and ascitic tumor of sarcoma 180. It did not change significantly the immunoorgan weight but potentiated immune responses such as B cell differentiation and the release of superoxide anion from macrophages. These results suggest that the protein-bound polysaccharide of LC43 exhibited antitumor activities through the activation of immune-related cells and acted as an immunmodulator.

• Journal of Plant Biology
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• v.29 no.3
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• pp.197-205
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• 1986

Leaf mesophyll protoplasts from five cultivars of tobacco (N. tabacum L.) were cultured. The protoplasts did not survive in culture medium containing Murashige and Skoog inorganic salts for over 6 days. NH4NO3 and FeSO4.Na2EDTA concentration of Murashige and Skoog medium were toxic in tobacco leaf mesophyll protoplast culture. Therefore we investigated optimum condition in Murashige and Skoog medium. High plating efficiency was obtained by reducing the concentrations of NH4NO3 and FeSO4.Na2EDTA to 1/3 and 1/10, respectively, on the supplemented with 5$\mu$M IAA, 0.5 $\mu$M 2.4-D 5 $\mu$M BAP. Plants were regenerated from protoplast-derived calluses.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.1
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• pp.33-41
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• 1997

This study was carried out to establish simple and easy methods to judge the survival, senescence and death of the protoplasts in culture system by identifying the marker substance related to metabolic status of the cells. When rice and tobacco protoplasts were cultured in MS and KM-8P media containing 2,4-D or coconut milk ABA decreased especially in the media containing coconut milk, but GA$_3$, IAA and zeatin increased as the cultures progressed. The decrease of ABA and increase of zeatin was especially remarkable. When the supraoptimal amount of osmoticum (mannitol) was added to the culture media ABA decreased after a momentary increase, but other growth hormones slowly increased as the concentration of the osmoticum increased. Contents of individual hormones were contrasted when protoplasts rice and tobacco were cultured on the same medium containing 10mM super mine or NaCl. Tobacco protoplasts were more sensitive to NaCl stress and stopped protoplast division at the late stage of culture. Protoplast viability decreased greatly in 48 hours when the protoplast were at 32$^{\circ}C$ on a medium lacking several components. ABA content increased up to 10 days from incubation in negative proportion to the protoplast viability. On the other hand contents of other growth hormones, especially zeatin, decreased. The present results clearly showed that the contents of individual growth hormones in the plant protoplasts in culture varied sensitively in response to environmental factors that they are faced with. This indicates that the physiological states of the protoplast, such as survival, senescence or death can be simply judged based on the quantitative analysis of those hormones by ELISA.

• Korean Journal of Microbiology
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• v.24 no.3
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• pp.243-250
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• 1986

Protoplast formation and regeneration from wild-type and auxotrophic mutants of Candida pseudotropicalis CBS 607 as well as fusion between complementary mutants were carried out. Frequencies of protoplast formation from wild-type and histidine or adenine requiring mutants ranged from 96 to 100% whereas those from methionine or tryptophan auxotrophs were 52 and 72%, respectively. When bovine serum albumin(4mg/ml, BSA) was added to protoplasting buffer for cells of methionine or tryptophan auxotrophs grown in a medium supplemented with myoinositol(0.5mg/ml), 96-99 % of cells were converted to protoplasts. Protoplasts were regenerated at the frequencies ranging from 18 to 20%. However, the addition of BSA to protoplasting buffer and the supplement of myoinositol to a medium of cell growth doubled the regeneration rate except adenine auxotroph in which such an improvement was not observed. It was found that optimal concentrations of polyethylene glycol and $CaCl_2$ are 20% and 100mM while optimal pH and exposure time are 6.0 and 30min. The fusion frequencies between complementary mutants ranged from $1.5{\times}10^{-3}\;to\;8.8{\times}10^{-3}$ and were enhanced by the improvement in the rate of protoplast regeneration. When histidine auxotroph was fused with tryptophan mutant, several fusion products were obtained which were found to be in the state of aneuploid or diploid, judging from DNA content and the presence of a large nucleus in the products.

• The Korean Journal of Mycology
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• v.14 no.3
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• pp.215-223
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• 1986

Protoplasts of P. cornucopiae were reverted to normal hyphal growth and reversion frequency was $0.04{\sim}19%$. The complete medium stabilized with 0.6 M sucrose was most effective for regeneration of protoplasts. When hypertonic mushroom complete medium not containing agar was overlaid, regeneration frequency of protoplasts was the highest rate among the others of topagar. The protoplast reversion frequency and mycelial growth of P. cornucopiae were increased when various amino acids, nucleic acid components and vitamin compound were added to the hypertonic minimal medium. The relation between sources increasing reversion frequency and sources accelerating mycelial growth was similar in amino acids and nucleic acid components but it was different in vitamins. The protoplast reversion frequency showed the highest rate when all sources were added to the regeneration minimal medium. Microscopically, regeneration patterns of protoplasts showed formation of a bud-like structure, direct germination, yeast-like cell chain of the protoplast, and the production of both direct germ tube and yeast-like cell chain from a protoplast.