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.

Wheat breeding over the last 100 years has increased productivity by adapting genotypes to local conditions, but the genomic changes and selection signals that caused phenotypic change during breeding are essentially unknown. Studying and understanding human selection of multiple important genes controlling key phenotypic traits will promote wheat molecular breeding. A total of 1152 diverse global wheat materials were genotyped based on KASP markers from 47 genes controlling grain yield, grain quality, adaptation, and stress resistance. Significant phenotypic variations between landraces and modern cultivars were found in 11 adaptive and yield-related traits. Thirty-six improvement-selective favorable alleles, including 22 positive prolonged and 14 negative selection alleles, were identified through comparing frequency spectra. Sus1-7A-Hap-H, Sus1-7B-Hap-T, Sus2-2A-Hap-A, TGW6-A1a, Cwi-4A-Hap-C, vrn-A1, PHS1-PHS+ and Lr34+ were subjected to strong selection, and overwhelmingly strong selection had occurred before improvement selection at Psy-A1b, Psy-B1a or b, Psy-D1a and Cwi-5D-Hap-C. However, Rht-B1b, Rht-D1b and 1BL.1RS were rare or absent in Chinese landraces but present in modern Chinese cultivars and introduced accessions. Importantly, Lr68+, Fhb1+, Wx-B1b and Yr15+, currently existing at a low frequency, should be regarded as further major improvement targets in global wheat breeding. Gene flow analysis showed that introduced cultivars especially from the former USSR and Italy contributed to enriched genetic variation in modern Chinese cultivars. This work objectively reports human selection on favorable alleles of multiple crucial genes in Asia, Europe, North America and CIMMYT, and traces the distribution of important genes in global wheat for molecular breeding.

Summary: Plant‐specific WUSCHEL‐related homeobox (Wox) transcription factors (TFs) are crucial for plant growth and development. However, the molecular mechanism of Wox‐mediated regulation of thousand kernel weight (TKW) in crops remains elusive. In this research, we identified a major TKW‐associated quantitative trait locus (QTL) on wheat chromosome 5DS by performing a genome‐wide association study (GWAS) of a Chinese wheat mini‐core collection (MCC) in four environments combined by bulked segregant analysis (BSA) and bulked segregant RNA‐sequencing (BSR‐seq) of wheat grains exhibiting a wide range of TKWs. The candidate TaWUS‐like‐5D was highly expressed in developing grains and was found to strongly negative influence grain TKW and wheat yield. Meanwhile, the RNAi lines, CRISPR/Cas9‐edited single and double knockout mutants (AABBdd and AAbbdd), as well as the stop‐gained aaBB Kronos mutants, exhibited a significant increase in grain size and TKW (P < 0.05 or P < 0.01) and a 10.0% increase in yield (P < 0.01). Further analyses indicated that TaWUS‐like‐5D regulates TKW by inhibiting the transcription of sucrose, hormone and trehalose metabolism‐related genes, subsequently sharply decreasing starch synthesis in wheat grains. The results of this study provide a fundamental molecular basis for further elucidating the mechanism of Wox‐mediated regulation of grain development in crops.