Abstract
A simple enzymatic method has been developed for the isolation of protoplasts in workable quantities from radicles of 1—3 days germinating seeds of twelve plant species, including eight legumes (Cajanus cajan, Glycine max, Medicago sativa, Phaseolus aureus, P. mungo, Pisum sativum, Trigonella foenum-graecum, and Vicia fabs) and four members of the Cruciferae (Brassica alba (Sinapsis alba), B. campestris, B. napus and B. oleracea). Root protoplasts showed high division potential, with callus formation in most species tested except C. cajan and V. faba, where only a few cell divisions were observed. Plantlets were regenerated from root protop]ast-derived tissues via shoot formation in B. napus and B. oleracea, and somatic embryogenesis in M. sativa, demonstrating the totipotency of root protoplasts. Root protoplasts provide a useful experimental system for cultural studies and genetic manipulations, especially for those species which have previously been recalcitrant in culture. The advantages of root protoplasts are discussed, together with the problems still to be overcome with this system.
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