Innovative Protocols for Blackberry Propagation: In Vitro Cultivation in Temporary Immersion Systems with Ex Vitro Acclimatization

Optimized in vitro cultivation offers a sustainable solution to enhance blackberry (Rubus spp.) production while reducing pathogen contamination during propagation. This study developed and validated protocols for in vitro cultivation and ex vitro acclimatization of the Tupy, Brazos, and Kiowa cultivars at the Instituto Tecnológico Superior de Los Reyes, Michoacán. A 20 min treatment with 2% sodium hypochlorite (NaOCl) reduced contamination by below 10% and achieved explant survival rates exceeding 95%. Temporary Immersion Systems (TIS) with four to six immersion cycles of 5 min each maximized survival (above 95%) while minimizing necrosis and hyperhydricity and increasing fresh mass. Shoot development was significantly enhanced with 2 mg L−1 6-benzylaminopurine, and 1 mg L−1 indole-3-butyric acid promoted optimal root formation. Acclimatization success rates exceeded 90% in covered trays compared to significant losses in uncovered trays during early stages. These protocols enabled robust plant development and yields exceeding 10 t ha −1 during vegetative and reproductive stages, providing a scalable framework for sustainable blackberry production and broader applications in crop propagation.