Electrofusion of Plant Protoplasts and the Production of Somatic Hybrids

Bates, George W.

doi:10.1016/b978-0-08-091727-6.50019-8

Cited by 5 publications

(6 citation statements)

References 68 publications

(71 reference statements)

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“…Cultivar E41 gave a PE of 0.192% and 0.173% for G4. These values were consistent with those reported by others, where in many cases, less than 1% of protoplasts that were plated actually developed into colonies (reviewed by Bates 1992). Results determined that although initial yields varied (Table 3), protoplast responses in culture were similar.…”

Section: Optimum Culture Conditionssupporting

confidence: 81%

“…We expect the seven other kenaf cultivars would react similarly. Electrofusion frequencies have been reported to range from 40% to 70%, but less than 10% were binucleate due to the creation of chains of protoplasts as a result of dielectrophoresis (Bates, 1992). Although our fusion estimates were much lower, most observed fusion products appeared to arise from the fusion of two protoplasts instead of multi-cell fusions (Fig.…”

mentioning

confidence: 51%

“…Many somaclones have been demonstrated to be genetically distinct from the mother (original) plant and display unique or improved characteristics (Larkin & Scowcroft, 1981;Evans, 1989). Successful culture and regeneration of electrofusioninduced somatic hybrids (interspecific and intergeneric) have been reported for a number of species (reviews by Gaynor, 1986;Bates, 1992).…”

Section: Introductionmentioning

confidence: 99%

“…The normal range of fusion voltages for plant protoplasts had been reported to be single to multiple pulses at 0.1-2.0 kV cm -1 for 5-200 #see for square wave pulses (Gaynor, 1986;Bates, 1992;Sowers, 1993). Our high fusion voltage of 2.0 kV em -1 was most likely required because of the extremely short duration (3/tsec) of high voltage delivered to the protoplasts.…”

mentioning

confidence: 99%

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Protoplast isolation, culture, and fusion protocols for kenaf (Hibiscus cannabinus)

Reichert

Liu

1996

Plant Cell Tiss Organ Cult

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Protoplast isolation and culture protocols were developed for ten cultivars of Hibiscus cannabinus L. (kenaf). Leaves from seedling lines maintained in vitro were used as donor tissues. Optimal cell wall digestions were achieved with a combination of cellulysin (1.0%) and macerase (0.5%). Average yields ranged from 0.9× 105 to 5.9× 106 protoplasts g fw -l leaf tissue with viability estimates ranging from 53% to 87%. This protocol was ineffective for leaf tissue taken from plants grown in vivo. Protoplasts harvested from plantlets maintained in vitro produced rapidly growing calluses when plated in semi-solid medium after an initial culture in liquid medium. First cell divisions were observed within four to six days after initial culture in medium containing plant growth regulators 2,4-dichlorophenoxyacetic acid (1.4/~M) and kinetin (13.8/~M). An electrofusion protocol which did not significantly reduce protoplast viabilities was developed for kenaf protoplasts. The maximum fusion frequency (4.6%) was obtained with an electrofusion voltage of 2.0 kV cm -I.

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Section: Optimum Culture Conditionssupporting

confidence: 81%

mentioning

confidence: 51%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Protoplast isolation, culture, and fusion protocols for kenaf (Hibiscus cannabinus)

Reichert

Liu

1996

Plant Cell Tiss Organ Cult

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“…Somatic hybrids between Solanum tuberosum germplasm and sexually incompatible Solanum wild species can be obtained through protoplast fusions (Bates, 1992). Moreover, the fusion between two diploid genotypes allows a return to the tetraploid level and, if fertile, the hybrid can be used as a parent in crosses within a classical potato-breeding programme.…”

Section: Introductionmentioning

confidence: 99%

Transfer of PLRV resistance fromSolanum verrucosum Schlechdt to potato (S. tuberosum L.) by protoplast electrofusion

et al. 2000

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SummarySomatic fusions between an accession of the diploid wild species Solanum verrucosum and a dihaploid S. tuberosum genotype were produced in order to incorporate resistance to potato leafroll virus (PLRV). In total 15 somatic hybrids out of 16 regenerants were Qbtained. Identification of hybrids was based on additive RAPD patterns, general morphological characteristics, chromosome numbers and chloroplast counts in stomata guard cells. A field trial was performed with the hybrids, their two parents and the control cultivar Kennebec to assess field performance and phenotypic variability. Yield parameters varied considerably among somatic hybrids. Some of the hybrids gave significantly higher yields, tuber numbers and tuber weights than both parents. Pollen fertility of hybrids ranged from 19 to 77%. Twelve hybrids were found to be resistant to PLRV.

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Comparative effects of fusion facilitators on electrofusion attributes of N. tabacum mesophyll protoplasts

Matibiri

Mantell

1995

Plant Cell Tiss Organ Cult

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Mesophyll protoplasts isolated from in vitro-grown Nicotiana tabacum L. shoots were subjected to electrofusion.Dielectrophoresis was induced by an AC field of 50 V cm -1 inter-electrode distance and 0.5 MHz oscillation frequency. Fusion was effected by two 0.7 kV cm-1 DC pulses, each of 50 Us duration, applied within one second of each other. Various chemical treatments were tested for their effects on dielectrophoresis efficiencies (percentages ofprotoplasts that made contact with at least one other protoplast under the AC field), fusion efficiencies (percentages of protoplasts participating in fusion events), cell lysis (percentages of protoplasts bursting during the electrofusion processes), overall viabilities of fusion products 24 h post-fusion and overall plating efficiencies 7 d post-fusion (percentages of fusion-derived cells that had undergone division). The various attributes assessed on the electrofusion of protoplasts in the control treatment, 10% mannitol, differed considerably for experiments carried out on different days. Relative to the control treatment, only the Ca 2+ treatments, and to a lesser extent lipase treatment reduced dielectrophoresis efficiencies. Polyamines, cytochalasins and Ca 2+ treatments significantly reduced cell lysis percentages. All electrofusion facilitators tested (except for spermine at 150 mg 1-1, the cytochalasins B and D, and Ca 2+ treatments) increased fusion efficiencies to more than 1.5 times those obtained with the standard 10% mannitol electrofusion medium. Ca 2+ treatments increased overall viabilities of fusion products by more than 1.5 times. With the exception of the prostaglandins, lecithin and CaC12 treatments, overall plating efficiencies were reduced by treatment of protoplasts with fusion facilitators. Substantial increases in overall plating efficiencies over those observed in the control treatment were obtained using prostaglandin F2a, lecithin and CaC12.2H20 treatments. The implications of the results are discussed.

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Electrofusion of Plant Protoplasts and the Production of Somatic Hybrids

Cited by 5 publications

References 68 publications

Protoplast isolation, culture, and fusion protocols for kenaf (Hibiscus cannabinus)

Protoplast isolation, culture, and fusion protocols for kenaf (Hibiscus cannabinus)

Transfer of PLRV resistance fromSolanum verrucosum Schlechdt to potato (S. tuberosum L.) by protoplast electrofusion

Comparative effects of fusion facilitators on electrofusion attributes of N. tabacum mesophyll protoplasts

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