Protoplasts were isolated from root cortical explants of thirteen cultivars of Pisum sativum. The roots were tested for starch in the cortical tissues. There was a considerable range in the average protoplast yields and the coefficient of variation of those yields from the thirteen cultivars. Cultivars with a higher incidence of starchy roots had lower average explam protoplast yields and greater variation in their protoplast yields. Significantly higher protoplast yields were observed from explants with no starch or few starchy cells. Increasing the germination time of the peas increased the starch content in the seedling roots and decreased the protoplast yields.
Mitotic figures of diploid, tetraploid, octaploid and 16-ploid nuclei were observed in cultures of pea root protoplasts whose initial DNA content was apparently 2C and 4C. The distribution of these mitotic figures in the different ploidy levels paralleled the distribution of mitotic figures in the culture of intact root explants and may be related to the hormonal stimulation of mitoses in these cultures. The patterns of the time course of both DNA synthesis and cell division in the protoplast cultures were similar to such patterns observed in the culture of intact root explants, although longer lag periods were observed in the protoplast cultures. Mitotic abnormalities including both chromosome breakage and spindle disfunction were observed in protoplast cultures. A large portion of the cell pairs derived from mitoses (27 % in one experiment) contained Feulgen-positive micronuclei. An accumulation of an as yet unidentified differentiation product termed dense cytoplasmic protoplast derivative was observed. Some of the conditions influencing the development of these derivatives are reported.
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