The present study investigated the effect of different salts on cotton shoot regeneration and transformation efficiency. Two-day-old germinating embryos of a local cotton cultivar (SG-125) were pretreated with 50 mM each of NaCl, CaCl2, and KCl for 60 min. The embryo explants were transformed by cocultivation with Agrobacterium tumefaciens strain LBA 4404 harboring a binary plasmid pTF101.1 that carried the insecticidal gene (cry1Ac) under control of wound-inducible promoter (AoPR1) and bilanafos acetyl reductase (bar) gene for plant selection. The salt-pretreated embryos showed maximum response on regeneration MS medium containing 0.50 mg/L 6-benzylaminopurine (BAP) and 0.10 mg/L indole-3-butyric acid (IBA), also supplemented with 5 mg/L bialaphos for in vitro screening of the transformed plantlets. The primary transformants were further screened by molecular techniques for integration and expression of the introduced gene. Maximum transformation efficiency (1.10%) was noted on KCl-treated explants compared to nontreated (control) explants. In conclusion, pretreatment of explants with 50 mM KCl for 60 min induced positive effects and triggered shoot regeneration in primary cotton transformants.