Background: Homoeologs are defined as homologous genes resulting from allopolyploidy. Bread wheat, Triticum aestivum, is an allohexaploid species with many homoeologs. Homoeolog expression bias, referring to the relative contribution of homoeologs to the transcriptome, is critical for determining the traits that influence wheat growth and development. Asymmetric transcription of homoeologs has been so far investigated in a tissue or organ-specific manner, which could be misleading due to a mixture of cell types. Results: Here, we perform single nuclei RNA sequencing and ATAC sequencing of wheat root to study the asymmetric gene transcription, reconstruct cell differentiation trajectories and cell-type-specific gene regulatory networks. We identify 22 cell types. We then reconstruct cell differentiation trajectories that suggest different origins between epidermis/cortex and endodermis, distinguishing bread wheat from Arabidopsis. We show that the ratio of asymmetrically transcribed triads varies greatly when analyzing at the single-cell level. Hub transcription factors determining cell type identity are also identified. In particular, we demonstrate that TaSPL14 participates in vasculature development by regulating the expression of BAM1. Combining single-cell transcription and chromatin accessibility data, we construct the pseudo-time regulatory network driving root hair differentiation. We find MYB3R4, REF6, HDG1, and GATAs as key regulators in this process. Conclusions: Our findings reveal the transcriptional landscape of root organization and asymmetric gene transcription at single-cell resolution in polyploid wheat.

Modern wheat production comes from two polyploid species, Triticum aestivum and Triticum turgidum (var durum), which putatively arose from diploid ancestors Triticum urartu, Aegilops speltoides, and Aegilops tauschii. How gene expression during embryogenesis and grain development in wheats has been shaped by the differing contributions of diploid genomes through hybridization, polyploidization, and breeding selection is not well understood. This study describes the global landscape of gene activities during wheat embryogenesis and grain development. Using comprehensive transcriptomic analyses of two wheat cultivars and three diploid grasses, we investigated gene expression at seven stages of embryo development, two endosperm stages, and one pericarp stage. We identified transcriptional signatures and developmental similarities and differences among the five species, revealing the evolutionary divergence of gene expression programs and the contributions of A, B, and D subgenomes to grain development in polyploid wheats. The characterization of embryonic transcriptional programming in hexaploid wheat, tetraploid wheat, and diploid grass species provides insight into the landscape of gene expression in modern wheat and its ancestral species. This study presents a framework for understanding the evolution of domesticated wheat and the selective pressures placed on grain production, with important implications for future performance and yield improvements.

Plant biotrophic and hemibiotrophic pathogens modulate cellular distribution of host phosphoinositides and recruit PI(4,5)P2 to the plant-pathogen interfacial m

The widely recognized pleiotropic adult plant resistance gene Lr34 encodes an ATP-binding cassette transporter and plays an important role in breeding wheat for enhanced resistance to multiple fungal diseases. Despite its significance, the mechanisms underlying Lr34-mediated pathogen defense remain largely unknown. Our study demonstrates that wheat lines carrying the Lr34res allele exhibit thicker cell walls and enhanced resistance to fungal penetration compared to those without Lr34res. Transcriptome and metabolite profiling revealed that the lignin biosynthetic pathway is suppressed in lr34 mutants, indicating a disruption in cell wall lignification. Additionally, we discovered that lr34 mutant lines are hypersensitive to sinapyl alcohol, a major monolignol crucial for cell wall lignification. Yeast accumulation and efflux assays confirmed that the LR34 protein functions as a sinapyl alcohol transporter. Both genetic and virus-induced gene silencing experiments demonstrated that the disease resistance conferred by Lr34 can be enhanced by incorporating the TaCOMT-3B gene, which is responsible for the biosynthesis of sinapyl alcohol. Collectively, our findings provide novel insights into the role of Lr34 in disease resistance through mediating sinapyl alcohol transport and cell wall deposition, and highlight the synergistic effect of TaCOMT-3B and Lr34 against multiple fungal pathogens by mediating cell wall lignification in adult wheat plants.

Summary: Dissecting the genetic basis of complex traits such as dynamic growth and yield potential is a major challenge in crops. Monitoring the growth throughout growing season in a large wheat population to uncover the temporal genetic controls for plant growth and yield‐related traits has so far not been explored. In this study, a diverse wheat panel composed of 288 lines was monitored by a non‐invasive and high‐throughput phenotyping platform to collect growth traits from seedling to grain filling stage and their relationship with yield‐related traits was further explored. Whole genome re‐sequencing of the panel provided 12.64 million markers for a high‐resolution genome‐wide association analysis using 190 image‐based traits and 17 agronomic traits. A total of 8327 marker‐trait associations were detected and clustered into 1605 quantitative trait loci (QTLs) including a number of known genes or QTLs. We identified 277 pleiotropic QTLs controlling multiple traits at different growth stages which revealed temporal dynamics of QTLs action on plant development and yield production in wheat. A candidate gene related to plant growth that was detected by image traits was further validated. Particularly, our study demonstrated that the yield‐related traits are largely predictable using models developed based on i‐traits and provide possibility for high‐throughput early selection, thus to accelerate breeding process. Our study explored the genetic architecture of growth and yield‐related traits by combining high‐throughput phenotyping and genotyping, which further unravelled the complex and stage‐specific contributions of genetic loci to optimize growth and yield in wheat.

As with the majority of the hemibiotrophic fungal pathogens, the rice blast fungus Magnaporthe oryzae uses highly specialized infection structures called appressoria for plant penetration. Appressoria differentiated from germ tubes rely on enormous turgor pressure to directly penetrate the plant cell, in which process lipid metabolism plays a critical role. In this study, we characterized the MoPAH1 gene in M. oryzae , encoding a putative highly conserved phosphatidate phosphatase. The expression of MoPAH1 was up‐regulated during plant infection. The MoPah1 protein is expressed at all developmental and infection stages, and is localized to the cytoplasm. Disruption of MoPAH1 causes pleiotropic defects in vegetative growth, sporulation, and heat tolerance. The lipid profile is significantly altered in the Mopah1 mutant. Lipidomics assays showed that the level of phosphatidic acid (PA) was increased in the mutant, which had reduced levels of diacylglycerol and triacylglycerol. Using a PA biosensor, we showed that the increased level of PA in the Mopah1 mutant was primarily accumulated in the vacuole. The Mopah1 mutant was blocked in both conidiation and the formation of appressorium‐like structures at hyphal tips. It was nonpathogenic and failed to cause any blast lesions on rice and barley seedlings. RNA sequencing analysis revealed that MoPah1 regulates the expression of transcription factors critical for various developmental and infection‐related processes. The Mopah1 mutant was reduced in the expression and phosphorylation of Pmk1 MAP kinase and delayed in autophagy. Our study demonstrates that MoPah1 is necessary for lipid metabolism, fungal development, and pathogenicity in M. oryzae .