Effects of Sucrose and 6-Benzylaminopurine Concentrations on Shoot Regeneration and Vitrification in Aronia melanocarpa: Insights for Plant Tissue Culture Systems

Abstract

This study focuses on investigating the effects of sucrose and 6-Benzylaminopurine (BAP) concentrations on shoot regeneration and the development of vitrification in Aronia melanocarpa. Initially, the presence of BAP, in combination with varying sucrose concentrations, leads to a substantial increase in shoot number. The highest shoot formation is observed in the MS medium containing 3% sucrose and 5.0 mg/L BAP. Furthermore, sucrose concentration plays a crucial role in shoot growth, with higher concentrations promoting more extensive shoot development. However, when sucrose levels were at 3% combined with higher BAP concentrations (1.0 or 2.5 mg/L), an increased incidence of vitrification was observed over time. Interestingly, lower sucrose concentrations (1% or 2%) combined with 0.5 mg/L or 2.5 mg/L BAP initially delayed vitrification but eventually led to its occurrence. Microscopic analysis of leaf samples with varying levels of vitrification indicates significant differences in stoma characteristics, such as density and size, further confirming the detrimental impact of vitrification on cellular structures and physiological processes. The recovery of vitrified plants was evaluated using different growth media combinations. The absence of BAP in the medium led to higher recovery percentages without necrosis, while the addition of 0.5 mg/L BAP promoted shoot growth but potentially inhibited root development. During acclimatization, vitrified plants exhibited stunted shoot growth, shorter and narrower leaves, reduced root numbers and lengths, and decreased survival rates, particularly under lower humidity conditions. The cultivation period required for the recovery of the crop was determined to be six weeks under controlled greenhouse conditions.