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Plant Science Today (PST)

Research Articles

Early Access

Optimization of in vitro regeneration protocol for Fragaria × ananassa Duch. cv. Chandler and assessment of genetic fidelity through ISSR markers

DOI
https://doi.org/10.14719/pst.6585
Submitted
7 December 2024
Published
23-07-2025
Versions

Abstract

This research developed an effective in-vitro method for regenerating strawberry plants, facilitating the large-scale production of planting materials. Various vegetative parts, such as runner tips, nodal segments, shoot apices, stems and leaves, were used for direct organogenesis. These explants were grown on MS media enriched with varying concentrations of plant growth regulators. The highest shoot multiplication rate has been found to be 89.8 % and the maximum number of shoots per explant (4.3) were achieved using runner tips in culture media supplemented with 1.5 mg/L BAP and 0.1 mg/L of NAA. The optimal conditions included pH of 5.8, 0.6 % agar, 3000 lux light intensity, 16-hr photoperiod and an incubation temperature of 25 ± 2 °C. The optimal percentage of rooting (90.8 %) for the regenerated shoots was achieved on ½ MS medium enriched with 1.0 mg/L IBA along with highest number of roots per culture. Subsequently. all regenerated plants were hardened in a greenhouse with 85 % relative humidity. To confirm the genetic fidelity of the in vitro-derived plants, 12 ISSR primers were used, revealing monomorphic bands. The standardized protocol developed in this study is crucial for large-scale multiplication and can also aid in identifying any clonal variations or adulteration.

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