Optimization of protoplast isolation from tomato fruit (Solanum lycopersicum)

Protoplast isolation from plant tissues is important for plant breeding and genetic engineering programs. For this, the objective of the present study was to optimize protoplast isolation from tomato fruits. Four different treat-ments including water, CPW (101 mg/l KNO3, 1480 mg/l CaCl2.2H2O, 246 mg/l MgSO4.7H2O, 27.2 mg/l KH2PO4& 0.16 mg/l KI), MS nutrient medium and Tris buffer containing 10, 15, or 20% sucrose, with 10 mgL-1pectinase,as protoplast isolation buffer were compared. Protoplasts were isolated by using all the treatments, however, the viability of protoplasts after 20 hours was different in different isolating solutions. While, isolated protoplasts by using water, CPW, and MS, did not survive after 20 hours incubation at room temperature, most of the pro-toplasts isolated by Tris buffer containing 15% sucrose remained intact after 20 hours. Since the transient gene expression in protoplasts requires incubation of transfected protoplasts for 8-16 hours, this protoplast isolation method can be used for transient gene expression studies in tomato

Keywords:

Protoplast , Tomato , transient gene expression