A cytological study was made of excised segments of pith of tobacco (ev. Wisconsin 38) grown in vitro on media containing various growth substances. Explants were cultured on a basal medium and on media containing kinetin, indoleacetic acid, gibberellic acid, and combinations of these three growth substances. Controls consisted of pith segments chemically fixed immediately after excision. The nuclei of explants on the basal medium and those on media containing the growth substances showed a response as early as 1 day after the beginning of the experiment. Nucleoli were indicators of initial stimulation in that they became swollen and displayed internal “vacuolation”, a known characteristic of active cells. These fatures were more pronounced by the second day for cultures grown on media containing growth regulators. Nucleoli were centers of nucleohistone staining as shown by the ammoniacal silver method. Continued and sustained growth of explants was possible only on media containing combinations of IAA and kinetin, or a combination of kinetin, IAA and GA. The greatest initial nucleolar stimulation occurred with IAA alone, even though subsequent growth did not take place. There was no nucleolar response of explants grown on abscisic acid al
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