Nodal segments from secondary branches of saplings of Phyllostachys bambusoides were inoculated in MS medium to assess the in vitro morphogenic response of leaf sheath through the induction to callogenesis by Picloram (4-amino-3,5,6-trichloropicolinic acid) at different concentrations of carbohydrate under the same conditions with presence or absence of luminosity. In our experiment, secondary explants were kept in MS medium containing 8.0 mg·L−1 of Picloram for the callus formation. Calluses were transferred in MS medium supplemented with sucrose, fructose and glucose (control, 2%, 4% and 6%). Results show that Picloram induced the callogenesis in leaf sheath. The secondary embryogenesis was formed in yellow-globular callus. The sucrose as carbohydrate source in the absence of light was more efficient to induce rhizogenesis. Glucose was more efficiency in the presence of light. Callogenic induction and further embryogenesis evidenced the competence and determination of leaf sheath cells.