Improved isolation and culture methods for cucumber mesophyll protoplasts

Improved methods for the isolation of large numbers of viable protoplasts from cotyledons and first leaves of two cultivars of cucumber, Cucumis sativus L. cv. Ashley and China are described. Short-term isolation methods (STM), and long-term isolation methods (LTM), using a variety of commercial enzymes are outlined. Liquid culture and subsequent solid culture of isolated first leaf protoplasts resulted in sustained cell division and eventual compact callus formation, which has been induced to form roots. No shoot or whole plant regeneration has, as yet, been demonstrated