The fidelity and reproducibility of a novel tissue culture-independent Agrobacterium tumefaciens-mediatedtransformation strategy was analyzed in cotton (Gossypium spp.) and the transmission study of the transgenes was doneby doing analysis at progeny level. Each 90 seedlings of Sahana and 70 of BCS 23-18-7 were used to transfer cry1Ac generespectively. Totally four and five plants of Sahana and BCS 23-18-7 were amplified with cry1Ac and npt-II specificprimers. However, all progenies of the T0 positive transformants failed to amplify in T1 generation. This rationalization ofthe results indicates the instability or failure of this method towards integrating the transgene at nuclear genome.