Temperature-Sensitive Variants of NICOTIANA TABACUM Isolated from Somatic Cell Culture

Temperature-sensitive variants of Nicotiana tabacum were isolated from a liquid suspension culture of somatic cells by a negative selection procedure, using bromodeoxyuridine and light. A total of nine such variants have been recovered, with an estimated rate of 2 x 10(-7) per cell division. The appearance of the variants at the permissive temperature varied from nearly wild type, white and friable, to brown, compact and slow growing. Two of the variants adapted from growth on solid medium to growth in a liquid suspension culture; these were further characterized for chromosome number, growth rate, cell death rate at the restrictive temperature, growth on nutritionally modified media, and RNA and protein synthesis. The variants have been placed on regeneration media, and one of them has produced plantlets. Leaves from a plantlet have been placed on callus-inducing media, and the resulting callus displayed the temperature-sensitive phenotype.