RECOVERY OF A PANTOTHENATE AUXOTROPH FROM A CELL-SUSPENSION CULTURE OF DATURA-INNOXIA MILL

A predominantly haploid (n = 12) suspension culture of Datura innoxia Mill. was used to devise methods for the isolation of auxotrophic cells strains. The method employed the filtration of cultures to produce suspensions consisting mainly of single cells. After mutagenesis with ethane methane sulphonate (EMS), filtered cell suspension were cultured in liquid medium for 8 days and then plated at low density and with high plating efficiency onto solid medium. Non-selective testing of some of the resulting calli revealed a colony which both in liquid and on solid media required pantothenate for growth. Although the pantothenate-requiring cell line appears to be largely tetraploid (4n = 48), date are presented to show that it could have arisen from single, haploid cells. © 1979.