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The effect of chemical facilitators on the frequency of electrofusion of tobacco mesophyll protoplast

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Abstract

Trypsin, pronase, protease, dispase, spermine, spermidine, and DMSO were characterized for their effect on the frequency of electrofusion of tobacco mesophyll protoplasts. Protease (Boehringer Mannheim) and Sigma pronase (1.26 mg/ml; 15 min incubation) increased the fusion frequency from 7% (control) to 20.7% (2.9X increase). Following protease and pronase were trypsin (2.8X), spermine (2.4X), dispase (2.1X), DMSO (2.0X), and spermidine (1.4X). BM Protease and polyamines caused the least amount of damage, followed by DMSO and trypsin (26% and 24% decrease in viability respectively), pronase (41%) and dispase (57%). Callus formed from all but dispase-treated protoplasts. Shoots regenerated from calli of all but trypsin-treated protoplasts.

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Abbreviations

BA:

6-Benzylaminopurine

BM:

Boehringer Mannheim

DMSO:

dimethyl sulfoxide

FDA:

fluorescein diacetate

FM:

protoplast fusion medium

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid, sodium salt

MES:

2-(N-morpholino)ethanesulfonic acid

MS:

Murashige and Skoog (1962)

NAA:

α-Naphthaleneacetic acid

PEG:

polyethylene glycol

SE:

standard error of the mean

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Author notes

  1. Suzanne C. McCarthy

    Present address: Aphton Corporation, P.O. Box 1049, 95695, Woodland, CA, USA

Authors and Affiliations

  1. Plant Genetics, Inc., 1930 Fifth Street, 95616, Davis, CA, USA

    Steven E. Ruzin & Suzanne C. McCarthy

Authors
  1. Steven E. Ruzin
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  2. Suzanne C. McCarthy
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Communicated by G. C. Phillips

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Ruzin, S.E., McCarthy, S.C. The effect of chemical facilitators on the frequency of electrofusion of tobacco mesophyll protoplast. Plant Cell Reports 5, 342–345 (1986). https://doi.org/10.1007/BF00268597

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  • DOI: https://doi.org/10.1007/BF00268597

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