A meristem micropropagation system was developed to produce Agrobacterium -free muscadine grape. Meristems were cultured on a modified Woody Plant Medium (mWPM) supplemented with 0.45 μm BAP. After 2 weeks, cultures were transferred to mWPM containing 8.92 μm BAP to enhance shoot proliferation. Propagules were subsequently subdivided and transferred to fresh medium at 2- to 4-week intervals. New shoots were excised and inserted in mWPM supplemented with 0.57 μm IAA to promote root formation. This method has been successfully used to produce Agrobacterium -free plants of muscadine cultivars Carlos, Doreen, Jumbo, Magnolia, and Sterling for research purposes and for a foundation planting in Mississippi. Chemical names used: benzylaminopurine (BAP); indole3-acetic acid (IAA).