Grain length, width, and thickness were measured using a vernier caliper from three randomly chosen, fully filled grains. The 1,000-grain weight was determined by weighing three replicates of 1000-grain samples independently on an electronic balance.
Endosperm was collected and then ground to a powder using a grinding mill. Finally, these powders are passed through a 50 mesh screen. The starch and amylose contents were measured using starch assay kits according to the manufacturer’s instructions (K-TSTA and K-AMYL; Megazyme). The Blue Value (BV) and maximum absorbance (λmax) of amylopectin were measured as described by Fu and Xue (2010) (link). To determine the amount of soluble sugars, 50 mg of endosperm powder was washed twice in 80% (v/v) ethanol at 80°C for 40 min and assayed using anthrone reagent (Wang et al., 2013 (link)). To determine the amount of reducing sugars, 100 mg samples of powdered endosperm were boiled in a small amount of distilled water for 30 min in a 25 ml volumetric flask, cooled to room temperature, diluted to volume with distilled water, and assayed using the DNS colorimetric method (Miller, 1959 (link)).
Endosperm was collected and then ground to a powder using a grinding mill. Finally, these powders are passed through a 50 mesh screen. The starch and amylose contents were measured using starch assay kits according to the manufacturer’s instructions (K-TSTA and K-AMYL; Megazyme). The Blue Value (BV) and maximum absorbance (λmax) of amylopectin were measured as described by Fu and Xue (2010) (link). To determine the amount of soluble sugars, 50 mg of endosperm powder was washed twice in 80% (v/v) ethanol at 80°C for 40 min and assayed using anthrone reagent (Wang et al., 2013 (link)). To determine the amount of reducing sugars, 100 mg samples of powdered endosperm were boiled in a small amount of distilled water for 30 min in a 25 ml volumetric flask, cooled to room temperature, diluted to volume with distilled water, and assayed using the DNS colorimetric method (Miller, 1959 (link)).
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