# Improvement of plant regeneration through the electroporation of colt Cherry (Prunus avium x pseudocerasus) isolated protoplasts

S.J. Ochatt, P.K. Chand, E.L. Rech, M.R. Davey, J.B. Power

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S.J. Ochatt, P.K. Chand, E.L. Rech, M.R. Davey, J.B. Power. Improvement of plant regeneration through the electroporation of colt Cherry (Prunus avium x pseudocerasus) isolated protoplasts. Electronique et pilotage des plantes, Moët-Hennessy., Sep 1987, Monaco, France. 1 p. hal-01600683

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Moët-Hennessy

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ELECTRONIQUE

ET

PILOTAGE

DES

PLANTES
IMPROVEMENT OF PLANT REGENERATION THROUGH THE ELECTROPORATION OF COLT CHERRY (Prunus avium x pseudocerasus) ISOLATED PROTOPLASTS

OCHATT S.J., P.K. CHAND, E.L. RECH, M.R. DAVEY and J.B. POWER. Plant Genetic Manipulation Group, The University of Nottingham, University Park, Nottingham, NG7 2RD, UNITED KINGDOM.

The high-voltage pulse transfection technique known as electroporation has been used in the past, among others, to transfer foreign genetic information into mammalian cells, to transform bacterial spheroplasts, and for the transformation of higher plant protoplasts, but in spite of this a knowledge of the possible action effects of electrical treatments on cell growth and development is scarce and restricted only to animal cell systems. The experiments to be reported in this contribution were aimed at examining the effects of electromanipulating isolated plant protoplasts and the influence of such a treatment on their viability and subsequent responses in culture.

Electric field pulses, ranging from 250 V to 2000 V and of 10 to 50 Usec duration, were assessed for their effects on the performance, in culture, of isolated protoplasts of five different higher plant species, eg *Glycine canescens*, *Solanum dulcamara*, *S. viarum*, *Pyrus communis* and *Prunus avium* x *pseudocerasus*. Three successive exponential pulses of voltages between 250 V and 1000 V caused a relatively small decrease in protoplast viability whilst they simultaneously favoured an earlier onset of division and enhanced the plating efficiency. A threshold for membrane lysis and protoplast collapse was found at voltage pulses beyond 1250 V. The percentage of protoplasts showing a sustained growth in culture to the microcallus stage and the rate of cell division as well were significantly higher for electro-treated protoplasts when compared to the controls.

Result showing a carry-over effect of the electroporation of the isolated cell suspension protoplasts of Colt cherry (Prunus avium x *pseudocerasus*) on their growth beyond the microcallus stage and on plant regeneration from such tissues are also reported. The electrical treatments enhanced both callus growth and plant regeneration responses of the protoplast-derived calli. The microcalli derived from protoplasts submitted to electric pulses at voltages of 250 V or 500 V showed the largest fresh weight gains between subcultures, for callus proliferation, and also the highest frequency or regeneration and number of shoots per callus. These shoots, in turn, showed a significantly more profuse root system than those derived from non-electropulsed protoplasts.

These observations are likely to be of considerable importance in the context of the introduction of foreign genetic information into higher plant protoplasts using electromanipulation technology. Any treatment that increases the extent and rate of protoplast division will be beneficial to the recovery of genetically modified plants. These results also show how electromanipulation techniques might have an additional practical application in enhancing the growth and regeneration responses of protoplasts which are generally regarded as less responsive, such as those of woody species.
Moët-Hennessy

CONFÉRENCE ET CONCOURS 1987
RÉSUMÉS DES COMMUNICATIONS

Monaco, 14, 15 et 16 septembre 1987

COMITÉ D'ORGANISATION

M. Jean-Pierre BARBE - Malicorne (France)

M. Alain DELOIRE - Épernay (France)

M. Gilbert DURAND - Toulouse (France)

Andrew GOLDSWORTHY - Londres (Royaume-Uni)

M. Jean-François LE MAITRE - Marseille (France)

M. Jean-Pierre MEGNIN - Paris (France)

SECRÉTARIAT

MOET-HENNESSY

Direction du Développement

30, avenue Hoche

75008 PARIS (France)

Tél. : (1) 45.63.01.01

Mme M.-C. COURAGEOT
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