• 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.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 Plant Biology
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• v.30 no.1
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• pp.1-9
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• 1987

Leaf mesophyll protoplasts of Nicotiana tabacum (nitrate reductase deficient mutant) were fused with cell suspension protoplasts of albino Petunia inflata in an electric field. Hybrid cell colonies were selected for nitrate reductase proficiency and chlorophyll synthesis. Five hybrid plant lines, regenerated from the selected calli lines, were analysed by electrophoresis, number of chromosomes and morphological characters. Somtic hybrid plants showed both parent patterns in the isozymesof isoleucine aminopeptidase and esterase. The hybrids had the expected chromosome number of 62 and exhibited an intermediate floral morphology when compared with the parents, but plant height and leaf arrangement were similar to N. tabacum.

• 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.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.169-174
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• 2005

The optimal culture conditions were studied for plant regeneration from mesophyll protoplasts of Solanum sisymbriifolium. Axenic seedlings of S. sisymbriifolium were used as a explant for protoplast culture. Many viable protoplasts were isolated by incubating leaf slices in an enzyme solution containing 0.25% Meicerase and 0.05% Macerozyme for 16 hr at $25^{\circ}C$ without shaking. Protoplast density of $5.0{\times}10^4\;ml^{-1}$ in Kao medium containing 5.0 mg/L NAA, 1.0 mg/L 2,4-D and 1.0 mg/L BA was optimal for colony formation. Most colonies were formed when protoplasts were cultured at $25^{\circ}C$ after initial culture at $30^{\circ}C$ for one week. On the MS agar medium with 1.0 mg/L zeatin, 38.4% of protoplast-derived calli differentiated shoots. These shoots rooted on 1/2MS medium with 5.0 g/L sucrose and 2.5 g/L gellan gum, and developed into whole plants.

• Journal of Plant Biology
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• v.15 no.3
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• pp.14-18
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• 1972

This paper includes a review on recent development on protoplast culture, regeneraton of plant from protoplast, and fusion of isolated protoplasts, and also describes the possibility of obtaining interspecific hybrid plants through asexual fusion of protoplasts of cells from distantly related plants which are not crossed by the ordinary sexual method.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.318-323
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• 1995

The effect of AF-toxin I produced by the strawberry pathotype of Alternaria alternata on the protein synthesis of susceptible strawberry protoplasts was examined by using the radiolabeled amino acids. The incorporation of the radiolabeled amino acids into newly synthesized proteins in the strawberry protoplasts was stimulated by the toxin treatment at relatively low concentrations (2.2$\times$10-11 to 2.2$\times$10-9 M), but not at higher concentrations (2.2$\times$10-8 to 2.2$\times$10-6 M). An one-dimensional SDS-polyacrylamide gel electrophoresis revealed no detectable differences in the proteins synthesized in both the toxintreated and untreated protoplasts. The susceptible strawberry protoplasts were treated with AF-toxin I and stained with Fluostain I to detect the extracellular polysaccharides. The toxin treatment induced the accumulation of extracellular polysccharides in a dose-dependent manner. These results indicate a transient activation of cellular metabolism in the susceptible cells by the toxin exposure.

• Journal of Plant Biology
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• v.28 no.3
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• pp.233-241
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• 1985

The isolation and culture of protoplasts from hypocotyl-derived calluses of Glycine max (L.) Merr. cv. Jangyeop were obtained by digestion for 6 hrs in an enzyme solution containing 3.5% cellulase, 1.5% macerozyme, 10% sorbitol and 0.1% CaCl2.2H2O at pH 5.8. Newly formed cell wall of protoplasts cultured in MS agar medium containing 10 $\mu$M $\alpha$-naphthaleneacetic acid (NAA) and 32 $\mu$M N6-benzylaminopurine (BAP) could be observed after 24 hrs culture. The first cell division of the protoplasts was observed after 3 days of culture; cell clusters after 2 weeks of culture. When transferred to solid media, the protoplasts formed cell clusters gave rise to proliferating calluses.