Somatic hybridization using a double mutant of nicotiana tabacum
Pental, D.; Hamill, J.D.; Cocking, E.C.
Heredity 53(1): 79-84
ISSN/ISBN: 0018-067X Accession: 006437522
Leaf mesophyll protoplasts of a nitrate reductase deficient, streptomycin resistant double mutant of N. tabacum were fused with cell suspension protoplasts of wild type N. rustica. Hybrid colonies were selected for nitrate reductase proficiency and streptomycin resistance. Green, actively proliferating colonies were recovered in the selection medium. Eleven of 20 green colonies, transferred to regeneration medium, produced plants. Regenerated plants were analyzed for their vegetative and floral characteristics, Fraction 1 protein polypeptide composition and leaf esterases. All the 11 regenerants were nuclear somatic hybrids possessing the chloroplast of the N. tabacum parent. By developing a double mutant with a negative (auxotrophic) and a positive (resistance) selection marker in a species of interest, this species can be hybridized by somatic cell fusion with wild type species lacking selectable markers.