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Bull. Spec. CORESTA Congress, Guangzhou 1988, p. 122, PP-4

N. Tabacum kanamycin resistant plants transformed by Agrobacterium tumefaciens

SORRENTINO C.; TONINI A.
Istituto Sperimentale per il Tabacco, Scafati, Italy
Gene transfer in plants can be obtained by DNA feeding of protoplasts or by natural vectors as A. Tumefaciens. In our research a gene for kanamycin resistance was introduced, as a selectable marker, in haploid plants of N. tabacum cv. Bright (057). Leaf disks were infected by an A. tumefacens binary vector carrying the nos-NPTII chimaeric gene. Calli formation and plant regeneration were performed in a Nitsch medium containing kanamycin 90 mg/l. New gene acquisition was confirmed on resistant calli and plants by Southern blot analysis using a nick-translated fragment of the binary vector as a probe. Plants were then diploidized by pit culture on a MS medium. Seeds so obtained were germinated on a MS medium (Hormone-free) containing kanamycin 90 mg/l. About 80% of germinated plants grew up well while the remaining 20% yellowed and died after a fow days. It is still to be investigated if and why the resistance kanamycin gene were lost from these plants.