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Journal of Experimental Botany, Vol. 51, No. 348, pp. 1237-1242, July 2000
© 2000 Oxford University Press

Original papers

Efficient intergeneric fusion of pea (Pisum sativum L.) and grass pea (Lathyrus sativus L.) protoplasts

P. Durieu and S.J. Ochatt1

INRA, Centre de Recherches de Dijon, Unité de Recherches en Génétique et Amélioration des Plantes, Laboratoire de Physiologie et Culture in Vitro, 21110 Bretenières, France

Large numbers of viable protoplasts of pea (Pisum sativum) and grass pea (Lathyrus sativus) were efficiently and reproducibly obtained and, for the first time, fused. Different procedures for fusion were compared, based either on electrofusion (750, 1000, 1250 or 1500 V cm-1), or on the use of macro or micro-methods with a polyethylene glycol (PEG 6000 or PEG 1540), or a glycine/high pH solution. Over 10% of viable heterokaryons were obtained, with PEG as the most efficient and reproducible agent for protoplast fusion (>20% of viable heterokaryons). Both the division of heterokaryons and the formation of small calluses were observed.

Key words: Protoplast fusion, Pisum sativum L., Lathyrus sativus L., grain legumes.


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