P.U. Kumara, A.G.K.K.M.W. Atapattu, H.M.I. Herath, D.L.C.K. Fonseka, K.K.I.U. Aruna Kumara, and F.M.M.T. MarikarOrchid is one of the most important cut flowers, grown in Sri Lanka for export and local market. One of the majors constrains in expanding the orchid cultivation is limited supply of quality planting materials. Micropropagation techniques can be successfully applied for mass propagation of orchids by the micropropagation is expensive thus increase the cost of production. Therefore, this research was undertaken for replacement of expensive agar for in vitro subculturing of Cattaleya seedlings. Two-month-old Cattleya seedlings were subcultures on Murashige and Skoog media (MS) as basal media. As gelling agents’ sago, semolina, corn flour, semolina + agar, and corn flour + agar was used. Control treatment was carried out with agar. Cultures were maintained inside the culture room. Number of leaves, number of roots, leaf length, leaf width and percentage of contaminations were recorded at four-week intervals. Sago and semolina + agar showed higher performance and low contamination percentage as alternative gelling agent in the MS media. The results of this study propose that sago, semolina, and corn flour can be used as an alternative gelling agent in MS media for subculture practices of orchids.