Quantification of ACC Content in Plant Tissues

ACC content was measured according to Zhang et al. with minor modifications. Powdered pericarp samples (1 g) were homogenized on ice in 10 ml 95% aqueous ethanol (v/v), and then centrifuged at 12000×g for 15 min at 4°C [2 (link)]. The supernatant was evaporated to dryness under vacuum at 40°C and the residue was dissolved in 5 ml distilled water. After oxidative conversion, ACC content was determined through ethylene production by gas chromatography (SHIMADZU, GC-2014C PF, Japan) as Lizada and Yang described [31 (link)]. ACC concentration of each sample was expressed in nmol∙g^-1 FW and the analysis was repeated three times.