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. 1986 Jan;80(1):117–121. doi: 10.1104/pp.80.1.117

Behavior and Viability of Tobacco Protoplasts in Response to Electrofusion Parameters

James A Saunders 1,2, Lee Ann Roskos 1,2, Sue Mischke 1,2,1, Mohammed A M Aly 1,2,2, Lowell D Owens 1,2
PMCID: PMC1075066  PMID: 16664564

Abstract

This investigation examines responses of protoplasts in a systematic and quantitative way to the various electrical treatments used to achieve electrofusion and their individual and cumulative effect on protoplast viability. Mesophyll and cell suspension protoplasts from two species of the same genera, Nicotiana tabacum and N. rustica var brasilia were used in these experiments. Optimal frequencies for alignment of tobacco protoplasts were between 500 kilohertz and 2 megahertz at 100 volts per centimeter. Variations in frequency and voltage of the alternating current (AC) field caused predictable movements of protoplasts within an electrofusion chamber. AC frequencies below 10 hertz or above 5 megahertz significantly decreased the viability of protoplasts in the fusion chamber as estimated by fluorescein diacetate staining 1 hour after treatment. Although the direct current (DC) pulse appeared to have a slight detrimental effect on protoplast viability, this effect was not significantly different from untreated control preparations.

Protoplasts from both leaf mesophyll cells and suspension cells were induced to fuse with one or more 10 to 30 microseconds DC square wave pulses of approximately 1 kilovolt per centimeter after the protoplasts had been closely appressed with an AC field.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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