Drought stress is one of the major abiotic stresses to wheat worldwide, with negative effects on wheat growth and yield. Assessing genetic variation and drought stress tolerance of key agronomic and physiological traits of spring wheat and screening germplasm resources for higher drought tolerance and yield stability are a prerequisite for developing new, better-adapted spring wheat varieties. This study evaluated nine important agronomic and physiological traits in 152 spring wheat cultivars under non-stress (NS) and drought-stress (DS) conditions. Under DS conditions, grain yield per plot (GYP) and grain weight per spike (GWE) were significantly reduced by 33.8% and 31.7%, and their drought-tolerance indexes (DIs) were only 0.66 and 0.69, respectively, indicating that GYP and GWE are the most susceptible traits to drought stress. The SPAD value of flag leave at flowering stage decreased by 13.9% under DS conditions, and the DI of SPAD was 0.86. In addition, DI-SPAD was significantly positively correlated with DIs of plant height (PH), grain number per spikelet (GPS), grain number per spike (GNS), GWE and GYP, indicating that the drought tolerance and yield of wheat are closely related to chlorophyll retention. Six wheat germplasm accessions were identified for their ability to sustain grain yield and improve drought tolerance simultaneously. These results provide insights into the genetic co-variation between grain yield and drought stress tolerance and provide a theoretical basis for the development of new wheat cultivars with excellent drought tolerance and high yields in the presence and absence of drought.

The branched spike phenotype is an important supernumerary spikelet trait of Triticum turgidum L. associated with the production of significantly more grains per spike, thereby offering a higher potential yield. However, the genetic basis of branch meristem (BM) development remains to be fully elucidated in wheat. TAW1, an ALOG (Arabidopsis LSH1 and Oryza G1) family gene, has been shown to function as a unique regulator in promoting BM development in rice. In this study, we found that the development pattern of the BMs of the branched spike in wheat was similar to the indeterminate BMs of rice. Moreover, phylogenetic analysis classified the ALOG genes into 12 groups. This family of genes was found to have evolved independently in eudicots and monocots and was evolutionarily conserved between wheat and rice as well as during wheat polyploidization. Furthermore, experiments revealed that TtALOG2-1A, a TAW1-homologous gene, plays a significant role in regulating the transition of indeterminate BM fate. Finally, large-scale RNA-sequencing studies and quantitative real-time polymerase chain reaction (qRT-PCR) experiments revealed that members of the TtALOGs may act upstream of the TtMADS22, TtMADS47, and TtMADS55 genes to promote indeterminate BM activities. Our findings further knowledge on BM development in wheat.

Wheat (Triticum aestivum L.) is one of the most important cereal crops worldwide, and its production is challenged by global climate change and a shortage of resources. The root system plays a vital role in uptaking water and nutrients and sensing soil environmental signals, and it has great potential to improve the final yield and stress tolerance of wheat. In order to further explore the genes regulating root development, this study focused on qRS-6D, located on chromosome 6D and spanning from 462,701,391 to 465,068,943, which was significantly associated with the total root length, root volume, root surface, and root fresh weight in our previous GWAS analysis. Firstly, its genetic effects were validated using an F₆ segregating population by comparing the root-related traits of homologous lines harboring the alternative haplotypes of this QTL. Then, the number of causal genes of this QTL was narrowed down to four with a transcriptome-wide association study. Additionally, qRS-6D has been demonstrated to have genetic effects on several yield- (kernel length, kernel width, and thousand-kernel weight) and plant structure-related traits (plant height, peduncle length, total tiller number, productive tiller number, flag leaf length, and flag leaf angle). Relatively, the frequency of the favorable haplotype increased with the wheat breeding practice. This study provides a reliable genetic locus to improve root development and structure and evaluate its application potential in wheat breeding improvement.

Awns play important roles in seed dispersal, protection against predators, and photosynthesis. The characterization of genes related to the formation of awns helps understand the regulation mechanisms of awn development. In the present study, the “double-awn” wheat 4045, which features super-long lemma awns and long glume awns, and an awnless wheat line, Zhiluowumai, were used to investigate QTLs or genes involved in awn development. QTL analysis identified three loci—Qawn-1D, Qawn-5A, and Qawn-7B—using a population of 101 4045 × ZLWM F₂ plants. Fine mapping with a total of 9018 progenies narrowed the mapping interval of Qawn-5A to an 809-kb region, which was consistent with the B1 locus, containing five genes on chromosome 5AL. Gene structure and expression analysis indicated that TraesCS5A02G542800 was the causal gene, which was subsequently verified by overexpression of TraesCS5A02G542800 in a “double-awn” wheat, Yangmai20. The retained “double-awn” phenotype of transgenic plants suggested that B1 represses the elongation but does not influence the emergence of the awns. Moreover, 4045 harbors a new allele of B1 with a 261-bp insertion in the promoter region and a lack of the EAR2 motif in the encoding region, which influences several important agronomic traits. In this study, we identify two novel QTLs and a novel allele of B1, providing new resources for exploration of awn development.

Deubiquitinating enzymes (DUBs) play essential roles in diverse plant biological processes, yet the ovarian tumor proteases (OTUs), a major DUB subfamily, have not been systematically characterized in wheat, and their functions in grain development remain unclear. Here, we identified 49 OTU genes (TaOTUs) in the hexaploid wheat genome and classified them into four subfamilies based on phylogenetic relationships, with nomenclature assigned according to homology. TaOTU6-7B was highly expressed during early and mid-grain development and was responsive to gibberellin and jasmonic acid. Its expression differed significantly between large-grain wheat Pindong34 (PD34) and small-grain wheat MY11847 at 7 and 11 days after flowering. To elucidate its function, we used CRISPR/Cas9 to generate loss-of-function mutants by knocking out the three homoeologs (TaOTU6-7A, -7B, and -7D). These mutants exhibited significantly increased grain width and weight relative to wild type. Moreover, TaOTU6-7B directly interacted with TaUBC13, whose expression was markedly elevated in the TaOTU6 knockout background, suggesting that TaUBC13 may positively regulate wheat grain size. Collectively, this study establishes the TaOTU gene family in wheat, reveals TaOTU6 as a negative regulator of grain width and weight, and provides valuable genetic resources and a theoretical foundation for high-yield wheat breeding.