Glyphosate-resistant crops have been a huge economic success for genetic engineering. The creating of new glyphosateresistantplants would increase the available choices for planting and lower the price of genetically modified crop seeds.A novel G6 gene from Pseudomonas putida that encoded 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) waspreviously isolated. The G6 gene was transfected into rice via Agrobacterium-mediated transformation. The transgenicrice obtained was confirmed by PCR, Southern, and Western blots. The lab experiment and field trials further confirmedthat the transgenic rice can survive glyphosate spraying at a dose of 8 g L-1. In contrast, conventional rice was killed ata weed control glyphosate spray dose of 1 g L-1. Altogether, the present study showed that the G6 gene works well in ricein vivo for glyphosate-resistance.