LAUSR

Summary Grain size and filling are two key determinants of grain thousand‐kernel weight (TKW) and crop yield, therefore they have undergone strong selection since cereal was domesticated. Genetic dissection of the two traits will improve yield potential in crops. A quantitative trait locus significantly associated with wheat grain TKW was detected on chromosome 7AS flanked by a simple sequence repeat marker of Wmc17 in Chinese wheat 262 mini‐core collection by genome‐wide association study. Combined with the bulked segregant RNA‐sequencing (BSR‐seq) analysis of an F2 genetic segregation population with extremely different TKW traits, a candidate trehalose‐6‐phosphate phosphatase gene located at 135.0 Mb (CS V1.0), designated as TaTPP‐7A, was identified. This gene was specifically expressed in developing grains and strongly influenced grain filling and size. Overexpression (OE) of TaTPP‐7A in wheat enhanced grain TKW and wheat yield greatly. Detailed analysis revealed that OE of TaTPP‐7A significantly increased the expression levels of starch synthesis‐ and senescence‐related genes involved in abscisic acid (ABA) and ethylene pathways. Moreover, most of the sucrose metabolism and starch regulation‐related genes were potentially regulated by SnRK1. In addition, TaTPP‐7A is a crucial domestication‐ and breeding‐targeted gene and it feedback regulates sucrose lysis, flux, and utilization in the grain endosperm mainly through the T6P‐SnRK1 pathway and sugar–ABA interaction. Thus, we confirmed the T6P signalling pathway as the central regulatory system for sucrose allocation and source–sink interactions in wheat grains and propose that the trehalose pathway components have great potential to increase yields in cereal crops.