Effect of Different Carbon Sources and Concentrations on in Vitro Propagation of Chestnut

Abstract

The in vitro propagation of chestnut is a crucial approach to address the limitations of traditional propagation methods. It encounters issues like insufficient rooting and multiplication. This research investigated the impact of sucrose, glucose, maltose and concentrations (20 g/L and 30 g/L) on the growth and biochemical profiles of the hybrid chestnut cultivar 'Marsol' (C. sativa x C. crenata). Essential parameters encompassed plant height, leaf area, callus size, chlorophyll content, enzymatic activity, and secondary metabolite composition determined using GC-MS analysis. For each carbon source and concentration, thirty-six biological replicates were analyzed. The findings underlined the essential function of sucrose, especially at 30 g/L, in enhancing plant height (5.30 +/- 0.04 cm), shoot multiplication (4.50 +/- 0.50 shoots per explant), and the coloration of leaves (minimum L value of 35.78 +/- 0.27, maximum SPAD value of 13.21 +/- 1.27). Glucose similarly promoted growth but to a slightly lower degree, whereas maltose continuously exhibited inferior performance. The callus's diameters, a crucial indicator of regeneration potential, were greatest with 30 g/L sucrose (1.664 +/- 0.26 cm width), highlighting its effectiveness in cellular differentiation. GC-MS analysis demonstrated various biochemical profiles affected by the carbon source. Sucrose treatments showed elevated concentrations of pyrogallol and hexadecanoic acid derivatives associated with stress tolerance and metabolic control. Furthermore, enzymatic tests revealed that the carbon source and concentration influenced phenol oxidase (PO) and catalase (CAT) activity, with sucrose producing ideal equilibria between oxidative stress and antioxidant responses. The findings provide critical insights for optimizing the nutritional medium to improve propagation efficiency for important chestnut cultivars/genotypes. Optimization of the carbon source and concentration are critical for in vitro propagation of chestnut (Castanea spp.). The study highlights that 30g/L sucrose, promotes optimal plant growth, biochemical balance, and stress resilience, establishing it as the optimal choice for enhancing propagation efficiency.