Transformation of Tagetes erecta L. with the GUS Reporter Gene Via Agrobacterium tumefaciens

Tagetes erecta L. (var. Marvel orange) a plant native to Mexico with traditional (used in religious ceremonies, as antiparasitic, insecticide, etc.) and industrial uses (flour of young flowers is used as food supplement in chicken and crustacean farms for its high content of carotenoids). Currently, there is great interest in achieving stable genetic transformation of this species with genes of agricultural interest; thus, the objective of this work was to optimize a protocol for stable transformation of T. erecta L. Shoot-tips from in vitro seedlings were transformed with Agrobacterium tumefaciens LBA4404 strain carrying the binary vector pCAMBIA 2301. Regeneration of transformed seedlings was performed in the presence of 50 mg & BULL;L-1 kanamycin. Stable genetic transformation of plantlets was demonstrated through positive GUS staining in leaves and flowers. PCR analysis detected the presence of a 600 bp fragment of the neomycin phosphotransferase II (NPTII) gene in the genomic DNA from regenerated seedlings. Transformation frequency was 40 %. To the best of our knowledge, this work is the second report of a successful stable transformation of T. erecta L. plants with GUS reporter via A. tumefaciens. This transformation protocol can be used to introduce genes in T. erecta L. to manipulate the isoprenoids biosynthetic pathways.