In vitro protocol for sterilization, callogenesis and shoot organogenesis for blueberry (Vaccinium corymbosum L.) var. ‘Blue ribbon’

Blueberry is a valuable fruit crop with significant antioxidant properties and economic importance. However, its breeding and propagation are limited by conventional methods. In vitro methods present promising options for the swift propagation and advancement of this crop. This paper focuses on establishing efficient protocols for in vitro sterilization, callogenesis, and shoot organogenesis in blueberry (Vaccinium corymbosum L.) var. ‘Blue Ribbon’. Effective surface sterilization of explants was achieved using 4% NaOCl for 4 min combined with 0.1% HgCl₂ for 2 min, reducing microbial contamination (19.97%) and improving explant survival (39.98%). The combination of 4 mg/L BAP and 0.75 mg/L NAA was most effective for callus induction, significantly reducing the callus formation period (5.66 ± 0.58 days) and increasing the induction percentage (97.96%). In terms of shoot development, 6 mg/L BAP was found to be optimal for quick shoot initiation (13.00 ± 1.73) and high shoot induction rates (81.39%). Additionally, 6 mg/L BAP and 6 mg/L TDZ were most effective in promoting the highest number of shoots per explant (4.66 ± 0.58 and 4.33 ± 0.58, respectively). This study provides a refined protocol for sterilization, callogenesis, and shoot induction.