Zak ERA8 (ENHANCED RESPONSE to ABA8) (Reg. No. GP-966, PI 669443) is a unique line derived from soft white spring wheat (Triticum aestivum L.) cultivar Zak that has increased seed dormancy but after-ripens within 10 to 16 wk. The goal in developing this germplasm was to use increased seed dormancy to improve tolerance to preharvest sprouting, a problem that can cause severe economic losses. This germplasm was developed by USDA–ARS, Pullman, WA, in collaboration with Washington State University. Zak ERA8was tested under experimental number 60.1.27.10. The ERA8mutation was generated by chemical mutagenesis followed by selection for the inability to germinate on abscisic acid (ABA) concentrations too low to inhibit wild-type Zak seed germination. The semidominant Zak ERA8 line has been backcrossed twice to wild-type Zak. Following the first backcross, Zak ERA8 showed similar morphological and grain quality traits to the original Zak cultivar.

Late maturity alpha-amylase (LMA) and preharvest sprouting (PHS) lead to elevated alpha-amylase in wheat (Triticum aestivum L.) grain. Risk of poor end-product quality due to elevated alpha-amylase is detected in the wheat industry using the Hagberg-Perten falling number (FN) method. In breeding programs, selection for PHS and LMA tolerance requires higher throughput methods requiring a smaller sample size than the 7 g required for the FN method. Specifically, LMA can only be screened only using detection of alpha-amylase activity or protein after cold treatment of individual wheat spikes at a specific stage of grain development resulting in very small samples (≤1 g). This study developed and evaluated a high throughput 96-well method for the Phadebas alpha-amylase enzyme assay for small wheat grain samples and compared this method to FN and the Megazyme Alpha-Amylase SD (Sprout Damage) Assay Kit performed on the automated Awareness Technology ChemWell-T Analyzer. In parallel, the efficacy of low-cost small-scale milling methods was evaluated relative to traditional larger scale mills. The Phadebas enzyme activity was highly reproducible and showed a strong correlation to the SD enzyme assay and FN method regardless of which mill was used to process the grain. The SD assay offers simpler standardization and calculation of enzyme activity, whereas the Phadebas assay offers higher sensitivity and lower expense. Both the 96-well Phadebas and automated Megazyme SD assays are suitable for alpha-amylase detection from small samples, and the use of low-cost coffee grinders to process small samples did not significantly impact assay performance.