<b>Background and Objective:</b> Tissue culture and thermotherapy were proved to be suitable in eliminating viruses of many plants. This study was designed in an attempt to produce virus-free Al-Taif rose plants (<i>Rosa damascena</i> Trigintipetala Dieck) through the practical application of the tissue culture approach and thermotherapy. <b>Materials and Methods:</b> Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay ( DAS-ELISA) and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) techniques were used to detect the presence of <i>Apple mosaic virus</i> (ApMV) and <i>Strawberry latent ringspot virus</i> (SLRV) in rose plant materials collected from Taif, KSA. RT-PCR was more sensitive than DAS-ELISA in detecting the 2 viruses. <b>Results:</b> Three different meristem-tip sterilization methods were compared and results revealed that treatment 3 (T<sub>3</sub>: 70% Ethanol for 1.0 min and 15% Clorox (Sodium hypochlorite 5.25%) for 10 min) was the most suitable as 97.78% of cleaned meristem tips survived. Meristem tips with different lengths were thermotherapy-treated for different durations. It was indicated that meristem tips of 0.5 or 1.0 cm and heat-treated at 37<sup>o</sup>C for four weeks gave the highest percentage of meristems that were able to differentiate into micro-shoots. <b>Conclusion:</b> RT-PCR detection of ApMV and SLRV revealed that using thermotherapy-treatment, for 4 weeks, of 0.5 cm long meristem tips was successfully applied to eliminate the 2 viruses in 92 and 96% of regenerated plantlets, respectively.
Keywords: ApMV; DAS-ELISA; Rose; SLRV; meristem; thermotherapy; tissue culture; virus-free.