Effects of Different Culture Media and Fe-EDDHA Applications on the In Vitro Propagation of Chestnut and Its Biochemical Profiles

Abstract

In addressing the limitations of conventional chestnut propagation, this study explored how varying Fe-EDDHA concentrations (0, 5.6, and 11.2 mg L-1) in different media (Murashige and Skoog, woody plant medium, and Gresshoff and Doy) affected the growth and biochemical traits of the hybrid chestnut variety 'Marsol' (C. sativa x C. crenata). Key factors, such as plant height, callus size, chlorophyll content, leaf color parameters, enzymatic activity, and HPLC-determined components, were assessed. The findings revealed that adding 5.6 mg L-1 Fe-EDDHA to the WPM notably improved plant height (3.89 cm) and leaf pigmentation, with the L value reaching a minimum of 33.29 and the SPAD value peaking at 10.67. In contrast, the GD medium promoted shoot proliferation (3.81 shoots per explant). The GD medium, when augmented with 5.6 mg L-1 Fe-EDDHA, yielded the most substantial callus diameter (1.71 cm width, 0.92 cm length), underscoring its efficacy in promoting cellular differentiation and regeneration potential. HPLC analysis revealed that Fe-EDDHA applications across different media affected biochemical profiles. The application of 5.6 mg L-1 Fe-EDDHA in GD medium promoted the proliferation of cells containing high levels of ascorbic acid, gallic acid, and 3,4-hydroxybenzoic acid. The findings provide valuable insights into optimizing culture media to enhance propagation efficiency for important chestnut cultivars or genotypes.