Novel starch synthase II (ssIIa) alleles were created in a soft wheat (Triticum aestivum L.) via ethyl methanesulfonate mutagenesis and combined by crossing to create four unique ssIIa triple mutant haplotypes. A range of starch amylose content was obtained from the ssIIa triple mutant lines (35.6–46.8%), with moderate increases in amylose content found in lines carrying two ssIIa null alleles and one partially functional ssIIa allele. Seeds from all ssIIa triple mutants had significantly higher amylose and protein contents but also lower starch content, kernel weight, and flour swelling power compared with their wild‐type sister lines. Seed starch amylose content was negatively correlated with individual kernel weight (r = –0.54), starch content (r = –0.85), and flour swelling power (r = –0.91) but positively correlated with grain protein (r = 0.78), demonstrating that unique ssIIa triple mutant null combinations confer defined levels of seed starch amylose, protein content, and kernel size. The ssIIa mutant lines also had a hard grain texture (≈86 single kernel characterization system units), whereas all parental material and wild‐type sister lines had soft grain texture (≈35). This change in grain texture was independent of Ha locus genotype, because all lines carried a functional (soft type) Ha locus. The ssIIa alleles and allelic combinations reported here demonstrate the ability to create defined levels of seed starch, amylose, and protein.
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