Tuning starch granule size distributions in durum wheat using genetic variation at a single locus

The size distribution of starch granules in wheat grains influences bread- and pasta-making quality, as well as nutritional properties. Here, we demonstrate that in durum wheat, wide variation in starch granule size distributions can be induced through missense mutations at a single genetic locus encoding the MYOSIN RESEMBLING CHLOROPLAST PROTEIN on chromosome 6A ( TtMRC-A1 ). We isolated 29 independent TILLING mutants in durum cultivar Kronos, each harbouring a different missense mutation that causes an amino acid substitution in the MRC protein. Compared to the B-type granule content of wild-type Kronos (24%), six of the missense lines had significant increases in B-type granule content (33-42%), although not to the extent observed in the mrc-1 mutant (58%) which carries a premature stop codon mutation. Notably, one missense line had significantly decreased B-type granule content (15%), demonstrating that mutations in TtMRC-A1 can achieve both increases and decreases in B-type granule content. In these lines, A-type granule size decreased as B-type granule content increased, and Rapid Visco Analysis on selected lines demonstrated that both B-type granule content and A-type granule size strongly correlated with pasting parameters (e.g., peak viscosity and pasting temperature). However, strong correlations between pasting properties and A-type granule size were still observed after removing most of the B-type granules via sieving, indicating that A-type granule size is the primary contributor to the observed variation in pasting properties. Overall, we demonstrate that mutations at TtMRC-A1 can greatly extend the range of granule size distributions in durum wheat, creating useful alterations in starch properties.