Development of an Efficient In vitro Propagation Protocol for Large Scale Production of Vitis vinifera L. cv. ‘Black Magic’

Abstract

This study aimed to standardize a reproducible protocol for direct and indirect micropropagation of the grapevine Vitis vinifera L. cv. ‘Black Magic’ using shoot tips, nodal segments, and young leaf tissues as explants. All experiments were conducted on MS basal medium. Direct shoot induction was found to be the most effective on 1.5 mg/l BAP supplemented medium, producing 86.67% response with 4.00 ± 0.55 shoots per nodal explants and 80% response with 3.20 ± 0.37 shoots per shoot tip explants. Maximum callus induction was obtained on 2.0 mg/l 2,4-D, producing 90% response in nodal explants and 75% in leaf tissues within 18-25 days. Organogenesis from callus was most successful on 1.5 mg/l BAP, producing 86.67% responsive calli with 14.20 ± 0.97 shoots per callus and 3.94 ± 0.41 cm in shoot length. Maximum shoot proliferation (92%) was achieved on 1.0 mg/l BAP + 0.5 mg/l NAA with 3% sucrose, yielding 17.89 ± 1.84 shoots per culture, 4.44 ± 0.29 cm shoot length, and 8.44 ± 0.58 leaves per shoot. The response was further enhanced by the addition of 5% coconut water, resulting in an average of 19.11 ± 1.60 shoots per culture, with a mean shoot length of 4.81 ± 0.71 cm and 9.00 ± 0.60 leaves per shoot. The number of shoots per culture increased from the 1^st to the 4^th subculture, reaching a maximum of 21.56 ± 2.77 shoots before declining in the 5^th cycle. Rooting was appeared to be the highest on half-strength MS medium with 3% sucrose and 1.0 mg/l IBA, achieving 88% rooting, 9.20 ± 2.46 roots per shoot, and a mean root length of 6.14 ± 0.62 cm. Rooted plantlets were transferred to polybags containing garden soil, sand, and compost (2 : 1 : 1), where 83.33% survival rate was recorded.

Plant Tissue Cult. & Biotech. 36(1): 269-281, 2026 (June)