• Korean Journal of Microbiology
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• v.19 no.4
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• pp.192-198
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• 1981

Protoplasts from Trichoderma koningii were fractioned at varing digestion periods. There were distinct differences between the fractionated protoplasts in CMCase activity and protein content ; namely, protein contents and CMCase activities in the early-protoplasts were higher than the late-produced protoplasts. Reversion frequencies of the early-produced protoplasts. Reversion frequencies of the early-produced protoplasts (0-1hr.) and the late-produced protoplasts (1-2.5 hr.)were 1.25% and 0.56%, respectively. From these results it was assumed that the early-produced protoplasts were more active in physiological metabolism. Meanwhile, in osmotically stabilized liquid medium two distinct reversion patterns of these protoplasts were found. In the first, normal germ tube was developed from the opposite side of the protoplast after the production of an aberrent tube. In the second, reversion occurred through germination of protoplast, but this patterns was uncommon. It is suggested that the the physiological and morphological heterogeneities in protoplast reversion are related to the heterogeneous origins of protoplasts from highly differentiated mycelium.

• Journal of Life Science
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• v.7 no.4
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• pp.282-288
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• 1997

To obtain a basic information on the development of Genus Viola, ultrastructure and electrofusion process between the two protoplasts from wild Viola callus cells and pansy mesophyll cells were observed with a scanning electron microscopy(SEM) and transmission electron microscopy(TEM). In the ultrastructural observation of wild viola callus protoplasts and pansy mesophyll protoplasts using SEM, their cell walls were removed completely. A knob-like formation was observed on the enlarge surface of viola callus protoplasts. On the surface of pansy mesophyll protoplasts net-like chloroplasts were observed. In SEM observation of pansy mesophyll protoplasts, chloroplasts devoid of membrane were observed on the surface the protoplasts. Pearl chain was formed by applying AC field of 200 V/cm at 1.0 MHz for 43 sec. The lysis of plasma membranes and fusion process occurred by applying a 1,600 V/cm DC pulse twice for 1 sec. After 1-2 hours of a DC pulse application, it was observed that the two protoplasts were fused completely into one cell. In TEM observation of the fused cell, many small vacuoles were located in the fusion area of the two protoplasts. Indeed, two distinct regions were observed during fusing process; in one region, a nucleus was found, while in the other region, both nucleus and nucleous were found.

• Journal of Plant Biology
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• v.15 no.4
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• pp.13-17
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• 1972

Rice callus was treated with 0.56M sucrose solution mixed with 5% pectinase and 10% cellulase, and the protoplasts isolated were transferred to 0.25 M sodium nitrate to induce protoplasmic fusion. Callus tissues were macerated well and degradation of cell walls also proceeded satisfactorily. When the protoplasts were transferred to sodium nitrate solution, many giant roundish protoplasts and some multilobed complex protoplasmic bodies were observed. Most of the fusions took place immediately after the protoplasts were transferred to sodium nitrate. Some multilobed protoplasts which failed to fuse in the initial stage took longer time, about two hours, to get completely fused and rounded-off. Multilobed protoplasmic bodies were invariably multinucleate, while giant round protoplasts had either several nuclei or had one nucleus of large size. Nuclear fusion, also, seemed to occur immediately after the protoplasts were transferred to sodium nitrate.

• Journal of the Korean Society of Tobacco Science
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• v.11 no.2
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• pp.233-240
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• 1989

This experiment was carried out to select of heterokaryon based on the different buoyant densities of protoplasts. Protoplats were isolated from cultured cells (calli) of Nicotiana tobacum(cv. BY4) and from mesophyll cells of N. glauca. The two types of protoplats were fractionated by centrifugation in an iso-osmotic (770 mOs/kg. H2O) density gradients condition. Major difference in the buoyant density exists between two types of protoplasts isolated from different cells. The mesophyll protoplasts were fractionated in the higher gradient interphases than that of callus protoplasts. The two types of fractionated protoplasts were fused with 40% polyethylene glycol (PEG), and the protoplasts treated with PEG were separated by centrifugation in the same density gradients condition. The heterokaryons were fractionated in the intermediate density gradients.

• Korean Journal of Microbiology
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• v.21 no.4
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• pp.213-220
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• 1983

Conditions for isolation of protoplasts from conidiospores of Trichoderma koningii ATCC 26113 were tested. Maximum production of conidial protoplasts was obtained by preincubation of conidiospores on liquid minimal medium for 8 1/2 hrs. and by reaction with cell wall lytic enzyme for 3 hrs. Among effective cell wall lytic enzymes (Driselase, p-Glucuronidase, Novozyme and Zymolyase), Driselase was the most effective one on the production of conidial protoplasts. The production of conidial protoplasts was also enhanced by addition of 2-Deoxy-D-Glucose $(25{\mu}g/ml)$ into liquid minimal medium. Over 70% of the initial swollen conidia, preincubated in liquid minimal medium supplemented with 2-Deoxy-D-Glucose $(25{\mu}g/ml)$, were converted to protoplasts by incubation with 2% (w/v) commercial lytic enzyme Driselase at $28^{\circ}C$ for 3 hrs. The reversion frequency of the conidial protoplasts was about 30 times (25-50%) higher than that of mycelial protoplasts (0.6-1.3%).

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.167-168
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• 2000

The potential application of protoplasts useful for studies such as physiology, morphology, genetic engineering, etc has led to the development of suitable methologies for isolation and manupilation of protoplasts from a wide variety of algae (Waaland et al., 1990; Reddy and Fujita, 1991; Chen and Chiang, 1994). Protoplasts technology to seaweeds depends large on the ability to produce viable cells capable of regenerating into whole plantlets (Wakabayashi et al., 1999). Though Capsosiphon fulvescens is one of the important economic seaweeds culturing in Korea, surprisingly protoplasts approach on this species has not been reported so far. Consequently we investigated the various aspects related to the protoplasts of Capsosiphon fulvescens in this study. (omitted)

• Journal of Plant Biology
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• v.32 no.4
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• pp.247-253
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• 1989

Protoplasts were enzymatically isolated from suspension culture of sweet potato. High yields of single protoplasts were produced from nonembryogenic cell aggregates. However, most protoplasts obtained from embryogenic cell clumps were spontaneously fused during enzyme treatment; a small portion of them remained single. Upon transfer to Murashige and Skoog's(MS) liquid medium supplemented with 0.1 mg/1 6-benzyladenine(BA) and 1 mg/12,4-dichlorophenoxyacetic acid(2,4-D), protoplasts from nonembryogenic cell aggregates sustained cell divisions to form cellus. Upon subculture onto MS media with 0.2 mg/12,4-D or without growth regulators, the callus did not give rise to any organs. On the other hand, first cell division of single protoplasts from embryogenic cell clumps was sporadically observed.

• Journal of Plant Biology
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• v.25 no.4
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• pp.181-188
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• 1982

Chloroplasts isolated from tobacco (Nicotiana tabacum L. cv. Virginia 115) leaves have been transferred into protoplasts of soybean (Glycine max Merr. cv. Jangyeop) suspension-cultured cells with the help of polyethylene glycol (PEG). The increased yield in protoplasts of chloroplast uptake was depended upon the concentration of both PEG 4,000 and PEG 6,000. The highest yield(36%) occurred at 50% of both PEG, and the yield was decreased above this concentration. The rate of uptake with the incubation time was highest at one hour, then decreased. The process of the chloroplast uptake into the protoplasts was similar with that of a protoplast fusion, except forming invagination during uptake.

• Journal of Plant Biology
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• v.29 no.1
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• pp.11-18
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• 1986

Protopasts were isolated from cultured cells of potato (Solanum tuberosum L.) tuber tissue. The ability of callus formation from the culture cells was higher in cultivars Dejima and Superior than in Shimabara and Irish Cobbler on Lam's medium. Therefore, the former was used as sources for protoplast isolation. Friable calli were transferred to liquid media and cells in exponential phase were used for protoplast isolation. In both of Dejima and Superior, the yield of protoplasts was high in the enzyme solution of 2% Onozuka cellulase and 1% macerozyme. Also, viability of isolated protoplasts was very good. Thus, it seems that these protoplasts would be applicable to various aims of research.

• Journal of Plant Biology
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• v.30 no.4
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• pp.287-298
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• 1987

The regenerative capacities of protoplasts isolated from potato (Solamum tuberosum L.) tubers and tobacco (Nicotiana tabacum L.) mesophyll tissues were examined, and then their intergeneric protoplast fusion was carried out. The potato tuber-derived protoplasts proliferated into the calli some of which showed rudimentary shoot-like structures, which had not been attempted before from tubers, while the tobacco protoplasts were regenerated into the whole plants. Intergeneric protoplast fusion between potato and tobacco was carried out and the heteroplasmic fusion products were formed. The first cell division of some of them was observed after 5 days of culture.