Development of an Efficient In Vitro Micropropagation and Microrooting Protocol from Half-Cotyledon Explants of Common Vetch (Vicia sativa L.)
Abstract
Common vetch (Vicia sativa L.) is an important forage legume; however, the lack of an efficient in vitro regeneration and microrooting protocol limits its utilization in rapid propagation and future breeding studies. The aim of this study was to develop an efficient in vitro micropropagation and microrooting protocol for the common vetch cultivar Kubilay 82 using abaxially cultured half-cotyledon explants. Seed surface sterilization was evaluated using commercial sodium hypochlorite solutions at six concentrations (10, 20, 40, 60, 80 and 100%) for three exposure periods (10, 15 and 20 minutes). Shoot regeneration was investigated on Murashige and Skoog (MS) medium supplemented with different combinations of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA), while regenerated shoots were rooted on MS medium containing 0.75 mg L⁻¹ indole-3-butyric acid (IBA). Seed sterilization was successfully achieved under all treatments without microbial contamination, and seed germination reached 100% under several sterilization combinations. Callus induction varied between 53.33 and 86.66%, whereas shoot induction ranged from 26.66 to 80.00%. The highest shoot induction (80.00%) and the greatest number of shoots per explant (13.66) were obtained on MS medium containing 0.20 mg L⁻¹ BAP without NAA. Microrooting percentage ranged from 60.00 to 86.66%, and the longest roots (9.96 cm) were obtained from shoots regenerated on medium containing 0.80 mg L⁻¹ BAP. The results demonstrated that BAP alone was more effective than BAP combined with NAA for shoot regeneration, while regenerated shoots exhibited high rooting capacity on IBA-containing medium. The developed protocol provides an efficient and reproducible regeneration and microrooting system for common vetch and represents a useful platform for future clonal propagation, genetic improvement, and biotechnology-based breeding studies.