Factors affecting the division frequency of pea pisum sativum mesophyll protoplasts

Jia S R.

Canadian Journal of Botany 60(11): 2192-2196


ISSN/ISBN: 0008-4026
Accession: 005440202

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Mesophyll protoplasts were isolated from 5 pea (P. sativum) cultivars ['Rondo', 'Century', 'Laxton's Progress', 'Trapper' and 'Afghanistan 72-HI-2'] and cultured in medium 8p, modified to test the effects of varieties, leaf ages, protoplast population densities, pH and plant hormones on relative division frequency. Protoplasts isolated from 'Rondo' had a much greater capacity for division than those from 'Century', 'Laxton's Progress', 'Trapper' or 'Afghanistan 72-HI-2'. The highest division frequency occurred in protoplasts isolated from the top expanded leaf of seedlings at the 3-, 4- or 5-leaf stage. The maximum frequency of division was obtained using a protoplast density of 5 .times. 104 to 1 .times. 105/ml and the 8p medium at pH 5.6 with 850 mg/l of CaCl2.cntdot.2H2O, 0.25-1.00 mg/l of 2,4-D and 2 mg/l of zeatin riboside. Gradually reducing the osmolality from 510 mosmol to 420 mosmol was essential to achieve a high frequency of cell division.