Experiments were conducted to develop a clonal propagation system for agarita (Berberis trifoliata Moric.). Actively growing agarita shoots were collected from a mature plant at the Texas A&M Univ. Research and Extension Center in El Paso and successfully established on a basal medium consisting of woody plant medium (WPM) salts and Murashige and Skoog vitamins, sucrose at 30 g·L–1, and 0.8% Phytagar supplemented with 11.1 μm BA. Cytokinins (benzyladenine, kinetin, and thidiazuron), subculture period, and age of cultures were tested. The optimal shoot proliferation conditions were WPM basal medium supplemented with 5.5 μm BA and a subculture period of 4 weeks. Culture age did not affect shoot proliferation but did affect rooting. Preliminary experiments with 1.0 μm NAA resulted in nearly 100% rooting of microshoots <6 months old. Shoots from 21-month-old cultures had to be placed on a cytokinin-free medium before successful rooting. On basal medium supplemented with NAA (5.4 μm), 68% of the microshoots rooted with an average of 1.2 secondary roots per microshoot. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA); 1-naphthaleneacetic acid (NAA); N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (thidiazuron or TDZ); 6-furfurlaminopurine (kinetin).