Differential RNA expression of Bmy1 during barley seed development and the association with β-amylase accumulation, activity, and total protein.

The objective of this study was to determine if developing barley (Hordeum vulgare L.) seeds had differences in β-amylase 1 (Bmy1) mRNA accumulation, β-amylase (EC 3.2.1.2) activity, β-amylase protein accumulation, and total protein levels during late seed development from genotypes with different Bmy1 intron III alleles. Two North American malting barley cultivars (Hordeum vulgare ssp. vulgare) were chosen to represent the Bmy1.a and Bmy1.b alleles and, due to limited Bmy1 intron III allele variation in North American cultivars, two wild barleys (Hordeum vulgare ssp. spontaneum) were chosen to represent the Bmy1.c and Bmy1.d alleles. Wild barleys Ashqelon (Bmy1.c) and PI 296897 (Bmy1.d) had 2.5- to 3-fold higher Bmy1 mRNA levels than cultivars Legacy (Bmy1.a) and Harrington (Bmy1.b). Levels of Bmy1 mRNA were not significantly different between cultivated or between wild genotypes. In all four genotypes Bmy1 mRNA levels increased from 17 to 19 days after anthesis (DAA) and remained constant from 19 to 21 DAA. Ashqelon and PI 296897 had more β-amylase activity on a fresh weight basis than Legacy and Harrington at all developmental stages. β-Amylase protein levels increased from 17 DAA to maturity in all genotypes. Total protein in grains from wild genotypes was significantly higher than cultivated genotypes at all developmental stages. Higher levels of total protein in Ashqelon and PI 296897 could explain their higher levels of β-amylase activity, when expressed on a fresh weight basis. When β-amylase activities are expressed on a protein basis there are no statistical differences between the wild and cultivated barleys at maturity.