An efficient in vitro propagation of clonal cherry rootstock Gisela-6 and validation of genetic stability through SCoT markers

Abstract

Gisela 6 or G6 (Prunus cerasus × Prunus canescens), a dwarfing rootstock, is vital in cherry cultivation due to its disease resistance and adaptability. However, traditional propagation methods face challenges in maintaining a consistent supply of quality G6 rootstocks due to variability in plant characteristics and disease susceptibility. To work over this, several combinations and concentrations of growth regulators were used to optimize the in vitro propagation of G6 rootstock, which was successfully multiplied following a 12-day period on MS medium, enriched with 0.5 mg/L BA and 0.3 mg/L GA₃. The highest successful shoot multiplication (1:6) on MS medium with 0.7 mg/L BA, 0.7 mg/L GA₃, and 0.1 mg/L IBA was observed after the fourth sub-culture. The two-step method that included growing on half-strength basal MS medium, following 48 h of culture on half-strength liquid MS medium supplemented with 0.8 mg/L IBA produced the maximum rooting (90.45%). Of the total plantlets, 60% got rooted, and survived after putting in pots, containing soil, sand, and FYM (1:1:1). The regenerated G6 plantlets were validated through 36 Start Codon Targeted Polymorphism (SCoT) markers, which revealed a high level of similarity (94%) between in vitro propagated and mother plants. The optimized in vitro propagation protocol, along with genetic stability assessment using SCoT markers, ensured consistent production of the true to type G6 planting material, addressing challenges in commercial cultivation, and advancing rootstock propagation for sustainable cherry production.