Studies on genetic fidelity of long term micropropagated culture derived plants of Ofra strawberry using molecular markers

Abstract

Availability of good plant material of a required cultivar ‘Ofra’ in a large quantity is a major limitation in expansion of strawberry cultivation. Keeping in view this problem, regeneration through in vitro culture has now become a viable and alternate method to conventional one. The formation of healthy shoots and higher rates of multiplication is one of the pre-requisite of an economically viable propagation. Somaclonal variations have been observed in the plants raised through tissue culture, which defeats the purpose of producing true-to-type plants. Therefore, need arises to study the genetic fidelity of tissue culture raised strawberry (Fragaria× ananassa Duch.) cv ‘Ofra’ plants. Runners of strawberry cv ‘Ofra’ used as explants and cultured on MS supplemented with 2mg/l GA3, 2.0 mg/l BA, 100 mg/l of meso inositol and 30g/l of sucrose gave best results for in vitro shoot multiplication. DNA from fresh, young and healthy leaves of 14 in vitro raised plants and mother plant of Strawberry cv “Ofra” was isolated following CTAB method. Random amplified polymorphic DNA and inter simple sequence repeat analysis were carried out with 15 and 20 primers out of which 7 and 14 primers produced three and 8 monomorphic bands respectively, for genetic stability studies of the regenerated plants. Comparison of the bands with the mother plant revealed the monomorphic nature and true to type clones. The above regeneration protocol of Strawberry (Fragaria × ananassa Duch.) cv ‘Ofra’ will be useful for micropropagation and genetic transformation studies.