Abstract: In order to study the subcellular localization of rice OsWAK1, the fusion protein of OsWAK1::GFP (green fluorescence protein) was constructed. A Agrobacterium tumefaciens strain EHA105 containing OsWAK1::GFP fusion was inoculated through the stomata using a 1-ml plastic syringe and by gentle pressure on the lower epidermal surface of tobacco leaves. In the experiment three tobacco species Nicotiana glutinosa, Nicotiana tabacum cv. Sam and Nicotiana benthamiana respectively were used. As a result, the green fluorescence emitted from Nicotiana glutinosa was detected, but no green fluorescence was detected from Nicotiana tabacum cv. Sam or Nicotiana benthamiana which indicate the difference of the fusion protein expression in the three tobacco species, the fusion protein was transient expressed in Nicotiana glutinosa, but not in the other two.