Production of Virus-Free Rose Plants using Meristem-Tip Culture and in vitro Thermotherapy

Abstract

Background and Objective: 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 (Rosa damascena Trigintipetala Dieck) through the practical application of the tissue culture approach and thermotherapy. Materials and Methods: 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 Apple mosaic virus (ApMV) and Strawberry latent ringspot virus (SLRV) in rose plant materials collected from Taif, KSA. RT-PCR was more sensitive than DAS-ELISA in detecting the 2 viruses. Results: Three different meristem-tip sterilization methods were compared and results revealed that treatment 3 (T3: 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 37oC for four weeks gave the highest percentage of meristems that were able to differentiate into micro-shoots. Conclusion: 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.