Given that glyphosate weed control is an effective strategy to reduce costs and improve economic outcomes of agricultural production in China,the development of glyphosate-resistant cotton holds great promise.Using an Agrobacterium-mediated transformation method,a new G2-aroA gene that encodes 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS)was transformed into cotton cultivar K312.The transgenic cotton plants were regenerated from a callus tissue culture via kanamycin selection.Ten regenerated cotton plants were obtained and allowed to flower normally to produce fruit.The results from polymerase chain reaction(PCR)and Southern and Western blot analyses indicated that the target gene was integrated into the cotton chromosome and was expressed effectively at the protein level.The glyphosate tolerance analysis showed that the transgenic cotton had a high resistance to glyphosate.Further,even cotton treated with 45.0 mmol L~(–1) of glyphosate was able to slowly grow,bloom and seed.The transgenic cotton may be used for cotton breeding research of glyphosate-tolerant cotton. Given that glyphosate weed control is an effective strategy to reduce costs and improve economic outcomes of agricultural production in China, the development of glyphosate-resistant cotton holds great promise. Using an Agrobacterium-mediated transformation method, a new G2-aroA gene that encodes 5 -enolpyruvylshikimate-3-phosphate synthase (EPSPS) was transformed into cotton cultivar K312.The transgenic cotton plants were regenerated from a callus tissue culture via kanamycin selection.Ten regenerated cotton plants were obtained and allowed to flower normally produce produce.The results from polymerase chain reaction (PCR) and Southern and Western blot analyzes that that target gene was integrated into the cotton chromosome and was expressed effectively at the protein level. The glyphosate tolerance analysis showed that the transgenic cotton had a high resistance to glyphosate. Further, even cotton treated with 45.0 mmol L ~ (-1) of glyphosate was able to slowly grow, bloom and seed.The transgenic cotton may be used for cotton breeding research of glyphosate-tolerant cotton.