Micropropagation of (Vitis vinifera L.) cultivar 'Sev Khardji' using biotechnological approaches and its impact on leaf quality

Abstract

Background: The grapevine (Vitis vinifera L.) is an anciently cultivated plant species with significant economic importance as a fruit crop worldwide. Many studies have revealed that it contains various bioactive compounds. In Armenia, all current, existing vineyards are based on planting materials propagated traditionally, which means they are not free of diseases. Yield losses in plants due to viral-related diseases can reach up to 90.0%. Meristem culture represents the sole method for obtaining virus-free planting material from infected plants.

Objective: The study aimed to establish an effective meristem culture technique for the Armenian aboriginal grapevine (Vitis vinifera L.) cv. 'Sev Khardji' and determine whether this technique influenced leaf quality.

Methods: Apical meristems served as explants for in vitro culture. The study employed various sterilizing agents and exposure durations to surface sterilize the explants. For shoot regeneration, the explants were cultured on full-strength Murashige and Skoog (MS) growth medium enhanced with various plant growth regulators (PGRs). In the process of in vitro root induction, different concentrations of two auxins, indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), were added to the ½ MS basal medium. The research investigated the levels of sugars, organic acids, vitamin C, and mineral content in fresh grape leaves.

Results: A remarkable explant survival rate of 90.0% was achieved through a co-treatment involving 70% ethanol (v/v) for 10 seconds and 1.0% sodium hypochlorite (NaClO) for 15 minutes. The highest shoot regeneration success rate (100%) was observed on growth medium containing 1.0 mg/l of 6-Benzylaminopurine (BAP), in combination with 0.5 mg/l of Kinetin (Kin) and 1.0 mg/l of gibberellic acid (GA₃). Additionally, a 100% success rate in root development was obtained using the nutrient medium enriched with 1.0 mg/l IBA. In vitro plants stored at 18±1°C, under a light intensity of 50 µmol/m²·s and a 12-hour photoperiod, remained viable for 14 months without requiring subculture. Rooted plantlets were acclimatized using a perlite and biohumus substrate mixture (2:1), achieving a survival rate of 92.0%. Improving propagation methods can enhance the biochemical qualities of grapevine leaves, potentially amplifying the health benefits associated with grapes.

Conclusion: For the first time, a successful micropropagation protocol has been developed for the ´Sev Khardji´ grapevine cultivar. This protocol considers the impact of phytohormones and their concentrations on plant regeneration and root formation. Beyond facilitating mass propagation, it serves as a valuable method for in vitro preservation. By improving the quality of grape planting material through biotechnological methods, this protocol has the potential to increase the health benefits linked to grape consumption. 

Keywords: ´Sev khardji´, grapevine, explant, in vitro propagation, functional foods, micropropagation, virus-free