2.5.4 Extraction and quantification of sucrose, fructose and glucose
Anthers were ground into fine powder in liquid nitrogen. 2 ml ethanol (80%) was added to 0.2 g of the ground tissue and incubated at 80 ⁰C for 30 minutes and centrifuged at 12000 rpm for 20 min at room temperature. The precipitate was re-suspended in 80% ethanol and re-extracted. The supernatants were pooled and incubated at 90°C until dryness, then 3 ml of ddH2O was added to the pellets and filtered using 0.45 µm microporous membrane. High-performance liquid chromatography (HPLC) analysis of sucrose, glucose, and fructose contents was performed from three biological replicates of each sample as described in a recently published paper (Pan et al., 2019).