Tortilla sales are projected to exceed 9.5 billion by 2014. However, currently no wheat cultivars have been identified that possess the intrinsic quality attributes needed for the production of optimum quality tortillas. Tortillas made with refined wheat flour low in dietary fiber (DF) are popular in the United States due to their sensory properties. This study explored the use of wheat lines (WL) possessing variations in high molecular weight glutenin allele sub-units (HMW-GS) for production of tortillas and also investigated the use of corn based resistant starches (RS), type II (RS2) and wheat based RS type IV (RS4) to increase DF in tortillas. Tortillas were made with 0-15 percent RS and 100 percent whole white wheat (WW). Flour protein profiles, dough, and tortilla properties were evaluated to determine the effects of the allelic variations and RS substitution on tortilla quality. Sensory properties of tortillas with RS were determined. Variations in HMW-GS composition significantly affected the protein quality and tortilla properties. Flour from WL possessing allelic combinations (2*, 17+18, 7, 2+12), (1, 17+18, 5+10), (2*, 17, 2+12) and (1, 2*, 17+18, 2+12) had 12.8-13.3 percent protein. These WL had extensible doughs and produced large diameter tortillas with superior (greater than or equal to 3.0) flexibility after 16 days compared to control. However, WL with (17+18 and 5+10) and (2*, 17+7, 5) produced extensible doughs, large, but less flexible, tortillas compared to control. WL with (2*,17+18,5+10) and (1,2*,7+9,5+10) produced smaller diameter tortillas, but with superior flexibility compared to control. RS2, WW, and cross-linked-pre-gelatinized RS4 (FiberRite) produced hard, less-extensible doughs and thinner tortillas compared to control, due to high water absorption. Cross-linked RS4 (Fibersym) dough and tortillas were comparable to control. 15 percent of RS2 and RS4 increase DF in control to 6 and 14 percent respectively, compare to control (2.8 percent DF). WW tortillas were less acceptable than control in appearance, flavor and texture, while tortillas with 15 percent Fibersym had higher overall acceptability than control. RS2 negatively affected dough machinability and tortilla shelf stability. However, 15 percent RS4 improved the DF in refined flour tortillas to meet FDA's "good source of fiber claim," without negatively affecting dough/tortilla quality.