Abstract
SummaryThis paper describes an efficient protocol for in vitro regeneration and genetic transformation from leaf explants of L. pennellii (Corr. D'Arcy). Shoot organogenesis was achieved on media with different combinations of indole-acetic acid (IAA) and kinetin (K). Regeneration ratios ranged from 50 to 90%. After co-cultivation with Agrobacterium tumefaciens, about 57% of infected explants formed calli in the presence of kanamycin, and 15% of these calli developed shoots within three weeks. Molecular characterization by polymerase chain reaction and Southern Hybridization confirmed the presence of the nptII gene and indicated the existence of plants with different number of copies. Histochemical assays for b-glucuronidase activity and rooting on kanamycin-containing medium indicated the expression of both genes. Reproduction of transgenic plants and sexual crosses with untransformed genotypes showed Mendelian segregation. The collection of L. pennellii transgenic plants will be an useful tool in experiments of asymmetric somatic hybridization with L. esculentum.
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