Kernel texture (grain hardness) is a key trait that influences the milling and baking quality of wheat (Triticum aestivum L.). Herein, hard red spring wheat varieties Butte86 and ND2603, possessing the Pina-D1b and Pinb-D1b alleles, respectively, were used as parents in the development of 132 recombinant inbred lines (RIL). The RIL population and its parents were phenotyped for single kernel characterization system (SKCS) and NIR kernel texture, grain protein content, test weight, SKCS weight, and SKCS diameter, and genotyped using next-generation genotyping-by-sequencing (GBS) technology, SSR markers and allele-specific sequence tagged site markers. Linkage maps for the entire RIL population and for two subsets of RILs, grouped on the basis of the Puroindoline-D1 alleles, were developed using a total of 695 markers. Multiple QTL mapping identified 10 QTLs for SKCS and NIR kernel texture with significant loci on 1AS, 1BS, 1BL, 5AL, 5BL, 6BL and 7BS. Of these, the 1BS QTL was associated with the Glu-B3 amplicon 691, and one of the 1BL QTL with Glu-B1. All QTL conferred an additive effect of ∼3–8 hardness units, a difference in kernel texture similar to or greater than the difference between the Pina-D1b and Pinb-D1b hardness mutations. •QTL analyses were conducted for hard red spring wheat RILs with different Puroindoline alleles.•A major QTL for kernel texture (hardness) was detected at the LMW Glu-B3 locus.•The Glu-B3 allele increased kernel hardness by ∼3–8 SKCS and NIR hardness units.