Genetic Homogeneity Analysis in Tissue Culture Raised Fragaria ananassa Duch. Revealed Through PCR Based Molecular Markers

The cultivated strawberry (Fragaria × ananassa) is globally renowned for its sweet fragrance, brilliant red hue, succulent texture, and its nutritional content and dietary fibres that promote health. Moreover, it stands as a high-value fruit crop of great significance to agriculture and local economies. The present investigation was carried out to develop an efficient micropropagation system for runner tips of virus tested strawberry cv. ‘Sweet Charlie’. The runner tips were sterilized with mercuric chloride (HgCl2) for 2 min and successfully established on MS medium supplemented with 0.5 mg/l BAP. The highest in vitro multiplication was obtained on MS medium containing 0.50 mg/l 6‑benzylaminopurine (BAP), 1.00 mg/l gibberellic acid (GA3) and 0.30 mg/l indole-3-butyric acid (IBA). The multiplied shoots were rooted on medium supplemented with different concentrations of IBA. The highest average root length and average number of roots were obtained on MS medium containing 0.50 mg/l IBA. The plantlets thus developed were successfully hardened in potting mixture of sand, soil and cocopeat (1:1:1). In order to obtain true to type plantlets with the aid of tissue culture, it is quite necessary to examine the genetic uniformity of in vitro raised plants. Hence, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were employed to determine the genetic uniformity of in vitro raised plants. In RAPD markers, OPA-05, OPA-07, OPA-08 and OPA-10 gave highly monomorphic bands. On the other hand, ISSR‑2, ISSR‑4, ISSR‑5, ISSR‑7 and ISSR3‑E showed complete monomorphism. The protocol thus developed for micropropagation of strawberry cv. ‘Sweet Charlie’ can be used over a long period of time without risk of somaclonal variations.