Effect of acetosyringone on agrobacterium-mediated transformation of cotton

Abstract

Genetic transformation method using Agrobacterium tumefaciens was developed for cotton plant species. Following the efficient regeneration of three cotton varieties, the effect of inclusion of acetosyringone in co-cultivation medium was measured. Subsequently, transformation was obtained by co- cultivation of 3 weeks old cotton apical shoot and A. tumefaciens strain LBA 4404 containing a plasmid harboring neomycin phosphotransferase and β-glucuronidase encoding genes. PCR analyses performed to identify the presence of marker gene (npt II) in the transgenic plants using primers used for amplification of a 700 bp fragment of the npt II gene showed that 87% of the transgenic plants selected for kanamycin resistance were transformed with the gene encoding β-glucuronidase. Routine transformation efficiency of cotton was established at 1.3%. The mean number of GUS positive apices was 67% higher when acetosyringone was included in the medium.