Plant Reproduction

Forward genetics is a powerful approach for gene discovery, but identifying causal mutations becomes difficult when mutants are maintained in heterogeneous populations with uncertain pedigrees. This is exemplified by classical tasselseed (ts) mutants, which have long served as a genetic model for studying sex determination and carpel suppression. Decades of repeated outcrossing to diverse inbred lines have created substantial genetic heterogeneity, limiting the effectiveness of conventional bulked-segregant analysis sequencing (BSA-Seq). To address this, we developed a BSA-Seq framework that integrates flexible experimental designs, multiple reference genomes, and complementary statistical methods tailored for genetically heterogeneous populations. Applying this framework revealed that reference genome selection is critical for mapping success and that Euclidean distance raised to the fourth power (ED4) outperformed homozygosity mapping (HM). Furthermore, the framework enables simultaneous mapping of multiple mutations within a single population, eliminating the need for additional mapping populations. Applying this framework to 26 ts mutant stocks from the Maize Genetics Cooperation Stock Center, we successfully mapped 24 mutants to genomic intervals containing known ts genes, while the remaining mutants mapped to distinct genomic intervals, defining novel candidate regions underlying carpel suppression. Together, these results demonstrate that historical mutant collections represent an underutilized resource for gene discovery and establish a generalizable mapping strategy for unlocking their genetic potential across diverse species.

Genomic selection (GS) and trait discovery in crops require reliable marker datasets, yet variant discovery remains constrained by the lack of standardised, high-confidence reference resources. Publicly available plant genomic resources are often based on outdated reference genomes, heterogenous pipelines, and generic variant hard-filtering thresholds derived from human studies. Consequently, implementation of the Genome Analysis Toolkit (GATK) Best Practices framework, including machine learning-based variant quality score recalibration (VQSR), remains challenging in crop genomes. Here, we present a crop-adapted framework for unsupervised variant quality assessment and VQSR enabling advanced SNP and insertion-deletion (INDEL) discovery without predefined high-confidence reference variant sets. Using Brassica napus as a model, we use the Illumina 60k Brassica SNP array to generate a reference-specific truth set for data pre-processing and independent evaluation of an unsupervised machine-learning approach for constructing cohort-specific VQSR resources. Gaussian mixture model (GMM)-based clustering of multidimensional variant annotation profiles identifies high-, intermediate-, and low-quality variant classes, providing flexible training resources for VQSR without requiring labelled training data. We further validate the framework in diploid Brassica oleracea and hexaploid wheat (Triticum aestivum) across cohorts differing in population size, genetic diversity, and genome architecture. Despite population- and species-specific differences in variant annotation profiles, GMM-based clustering consistently identifies informative variant quality classes and enables cohort-specific recalibration of both SNPs and INDELs. Our results demonstrate that unsupervised machine learning can extend GATK Best Practices to crop species lacking comprehensive genomic resources, providing a scalable approach for generating high-quality variant datasets for crop genomics and downstream breeding applications.

Petal coloration is a key ornamental trait in Cyclamen persicum Mill. and is closely associated with anthocyanin composition and relative accumulation, yet the major anthocyanins and candidate molecular factors associated with color variation remain unclear. In this study, anthocyanin profiling and transcriptome analysis were integrated to investigate three Cyclamen materials with distinct petal colors, BXK, FXK, and SHXK. Anthocyanin profiling showed clear separation among the three materials, indicating that petal color divergence was associated with differences in anthocyanin composition and relative abundance. Petunidin-, peonidin-, delphinidin-, malvidin-, and cyanidin-related derivatives exhibited distinct accumulation patterns. Transcriptome analysis revealed marked transcriptional divergence, with phenylpropanoid and flavonoid biosynthesis repeatedly enriched across comparisons, whereas isoflavonoid and flavone and flavonol biosynthesis were enriched in selected comparisons. Mapping differentially expressed genes onto the anthocyanin biosynthetic pathway revealed branch-specific expression patterns of structural genes, including CHS, F3H/F3'H, F3'5'H, DFR, and ANS. Notably, Cluster-26256.0, annotated as a putative anthocyanidin synthase gene, showed a progressive increase in expression from BXK to FXK and SHXK, consistent with increasing petal pigmentation. Several DFR-related transcripts also showed relatively high expression in SHXK. Co-expression analysis linked key anthocyanins, structural genes, and transcription factors, while qRT-PCR generally confirmed RNA-seq trends. These results suggest that late anthocyanin biosynthesis, particularly ANS-related transcriptional variation, may contribute to dark red petal coloration. This study provides a metabolite-transcript framework for Cyclamen petal coloration and identifies ANS-related genes as priority candidates for future functional studies and flower-color breeding.

BackgroundN6-Methyladenosine (m6A) regulates RNA fate in flowering plants, but the repertoire and expression patterns of m6A-related genes remain incompletely characterized in liverworts. Here, we characterized these gene families in the liverwort Marchantia polymorpha within a comparative framework spanning diverse plant lineages. ResultsA comparative survey of 119 plant genomes using sequence similarity and conserved-domain criteria identified 18 candidate m6A-related loci in M. polymorpha, comprising seven writer-related genes, five AlkB homolog genes, and six YTH- or CPSF30-related genes. Across the surveyed genomes, canonical writer components generally had low copy numbers, whereas AlkB homolog and reader-related families showed greater copy-number variation. Phylogenetic and domain analyses supported the family assignments of the M. polymorpha candidates. Eight of the 18 candidate loci showed detectable collinearity with Marchantia quadrata. Putative promoter analysis identified motifs annotated as hormone-, light-, and stress-responsive, with hormone-responsive motifs accounting for the largest share of predicted sites. Public transcriptome analysis revealed variation in candidate gene expression across developmental stages and tissues. Quantitative reverse-transcription PCR showed higher transcript abundance for all six YTH- or CPSF30-related genes in thalli than in gemmae or gemma cups. Cold, heat, osmotic, salt, and dark treatments elicited gene-specific expression responses. Darkness increased transcript abundance of several candidates, including MpMTA, MpALKBH1A, and MpECT2. A predicted protein-association network suggested links between subsets of the candidates and RNA-processing proteins. ConclusionsThese findings define a candidate m6A-related gene repertoire in M. polymorpha and identify tissue- and stress-responsive genes for targeted functional studies of RNA methylation in this liverwort.

Understanding how grafts form has fascinated scientists for over a century, and numerous transcription regulators involved in wound healing, callus formation, and grafting have been identified in model plants. However, the genetic architecture underlying grafting traits remains poorly characterized. To address this gap, we aimed to identify quantitative trait loci (QTLs) associated with grafting success rates and grafting-related traits (including callus formation, mechanical strength of the graft union, and post-grafting growth) in a grapevine rootstock mapping population, Vitis berlandieri x V. rupestris. Although no QTLs were identified for grafting success rate, QTLs were identified for component traits revealing a genetic architecture that would have remained hidden when considering grafting success as a single composite phenotype. By integrating QTL and transcriptome data, we identified regulators potentially mediating grafting-related traits. We then showed that one transcription factor that regulates callus formation and grafting in Arabidopsis potentially has a similar role in grapevine by silencing with exogenous double-stranded RNAs and that exogenous application of methyl jasmonate increases callus development at the graft interface. Candidate transcriptional regulators located within QTLs and evidence from gene silencing and hormone treatments suggest the conservation of regulatory mechanisms underlying wound-induced callus formation across plant species.

RNA-guided nucleases (RGNs), such as Cas9 from Streptococcus pyogenes (SpCas9), are widely used for plant genome editing. Previous surveys for off-targeting, the modification of unintended targets with similarity to the intended target, indicate high specificity of SpCas9 in plant cells. However, off-targeting has not been assessed for efficiency-optimized editing systems combined with extensive multiplexing, which might increase the likelihood of cleavage at unintended sites. We therefore analyzed Arabidopsis thaliana lines that had been extensively mutagenized using zCas9i and up to 29 gRNAs addressing >45 target sites over several rounds of editing. Genomes were sequenced by short- and long-read technologies, and genome-wide variants were catalogued. Our pipeline for variant calling reliably detected RGN-induced mutations at on-targets. When excluding these on-target modifications, variants were detected in edited lines at frequencies similar to those previously reported for spontaneous mutations. In further analyses, we did not find any evidence for an origin of these variants from RGN activity. Our data are thus consistent with high specificity of SpCas9. In contrast, we detected genomic reorganization events upon editing at two complex loci, RPP1 and RPP7, encompassing multiple homologous genes, and also identified an allele by WGS that had escaped detection by amplicon sequencing. We conclude that, while off-targets may efficiently be avoided by selection of specific gRNAs, on-target modifications may be more extensive than intended, especially at complex loci and/or during multiplexing.

The chloroplast is a semi-autonomous organelle with a compact genome encoding ~100 expressed genes. Numerous and widespread promoters coupled with inefficient transcription termination leads to a highly complex primary transcriptome that undergoes extensive RNA quality control orchestrated by RNA binding proteins and ribonucleases. Among the ribonucleases, only RNase J (RNJ) is absolutely required for plant germination and survival, likely because its deficiency leads to massive accumulation of antisense RNA which inhibits translation. As one approach to understanding the selectivity of RNJ for antisense RNA and noncoding regions, we created Arabidopsis lines overexpressing catalytically inactive versions of the enzyme designed to trap its substrates. Initially, transgenic plants appeared phenotypically normal. However, crosses used to generate homozygous transgenic lines yielded progeny that developed varying degrees of chlorosis within 2-3 weeks. Concomitantly, these plants exhibited chloroplast transcriptome alterations similar to RNJ-silenced lines, along with mRNA changes consistent with retrograde signaling. This chlorosis could be correlated to the likely decay of RNJ protein, but not its endogenous transcript. The result could be replicated by attempting to overexpress a wild-type version of RNJ, in which case even one transgene copy was sufficient to induce chlorosis. Our results suggest that RNJ is under protein expression level surveillance to prevent wholesale destruction of the plastid transcriptome.

Amaranthus tricolor is a climate-resilient orphan crop whose betalain pigments contribute to nutritional quality and stress tolerance. However, breeding efforts have traditionally relied on field yield trials due to the limited availability of genomic information and genetic resources. To address this gap, we first used the Illumina and PacBio sequencing platforms to assemble a novel reference genome of A. tricolor, using the Japanese cultivar Bayam. By combining this reference genome with a core collection developed in our previous study, we aimed to identify genomic regions associated with pigmentation-related traits across winter and summer seasonal environments. Betalains are antioxidant phytonutrients that contribute to abiotic stress tolerance, making the regulation of their biosynthesis under heat stress particularly relevant for developing climate-resilient crops such as A. tricolor. As a result, ATR1.0CH03_5131103, which is associated with ATR1.0ch03g000565, was identified under the winter seasonal environment. Meanwhile, transcriptomic profiles in this study revealed that ATR1.0ch03g000565 was differentially expressed under both control and heat stress conditions, suggesting that it may be associated with the regulation of CYP76AD1 expression under optimal conditions. Under the elevated-temperature treatment, transcript levels of CYP76AD1 and 5GT decreased in Am087, coinciding with lower betacyanin-dominated absorbance indices. cDOPA-like homologs were upregulated; however, pigmentation-related absorbance remained reduced. Further molecular experiments, such as CRISPR-Cas9 knockouts or overexpression studies, are necessary to validate the regulatory mechanisms of CYP76AD1 and 5GT under heat stress.

Breeding programmes choose genomic prediction models by accuracy rankings, and the metric behind a ranking decides which model wins. Selecting the best genotypes within a trial depends only on the order of the predictions, so a metric that guides the choice must be unchanged by any order-preserving change of the predictions and respond when their order is reversed. Of the 22 official metrics of the Genomes to Fields maize competition, one meets both conditions, the within-environment rank correlation; the within-environment Pearson correlation is protected against rescaling only; and twenty fail, including the root mean squared error that decided the 2022 ranking. Switching between the competitions two official metrics reverses 27 % of team pairs, and an exact decomposition shows why pooled metrics mislead: in three constructed panels, 94-95 % of the attainable square of pooled correlation is carried by the environment means. The accuracy difference a trial can resolve falls as about 3/{surd}N with its N genotype-environment cells: at least 0.03 at the competitions design and 0.04-0.12 once calibrated on real panels, against 0.003 between the first two teams; tests that two correlations differ declare far smaller differences, which do not reliably change selection. In soybean, where the two most accurate methods stand clear of most others by more than the trial resolves, ranking by either within-environment correlation selects better material, 98-99 % of the attainable selection gain in held-out environments against 13-34 % for error-magnitude rankings; in common bean and a Chinese maize population, where the leading methods lie within the resolvable gap, all rules pick material of similar value. GPverdict runs these checks on any programmes cross-validated predictions.

Zea mays (maize) flowering time is genetically determined and a critical yield determinant. Yet mechanistic understanding of maize flowering remains poor. Indeterminate1 (Id1), a zinc-finger transcription factor (TF), is a monocot-conserved master regulator of maize flowering. Epistasis between Id1 and the ZeaCentroradialis-Delayed Flowering1 (Zcn-Dlf1) inductive pathway partly explains ID1 floral control; however, the strong mutant id1- floral delay is not explained by this pathway alone. To better characterize Id1 actions, we performed single-cell assay for transposase-accessible chromatin and single nucleus RNA sequencing (scATAC-seq and snRNA-seq) comparing Id1+ and id1- developing leaves. These analyses reveal id1- chromatin remodeling via TEOSINTE BRANCHED1 CYCLOIDEA PROLIFERATING CELL FACTOR (TCP) and APETALA2/ETHYLENE RESPONSEFACTOR (AP2/ERF) transcription factors and provide candidate direct targets that include AP2/ERF genes. These candidate direct targets include the family of {beta}-glucosidase genes that lose expression in id1-. Unexpectedly, CRISPR/Cas9 {beta}-glucosidase edits produced plants that phenocopied terminal ear1- (te1-) mutants. This phenocopy prompted an investigation into the genetic relationship between id1-, te1- and flowering. Surprisingly, id1- te1- plants exhibited a synergistic floral delay, producing ~90 leaves before inflorescence production. Beyond highlighting hitherto unappreciated Te1 autonomous flowering roles, this genetic synergy raises the hypothesis that meristem leaf primordia cessation underpins maize flowering.

Despite the recent cloning of RLK-encoding resistance genes and the identification of a common point of hyphal arrest at the stomatal level in incompatible interactions, the molecular mechanisms leading to the establishment of wheat resistance against Septoria tritici blotch (STB) remain poorly characterized. Here, we use long-read RNA-sequencing data to compare the early transcriptomic responses of two wheat cultivars carrying major, isolate-specific resistances to Zymoseptoria tritici via differential expression, differential transcript usage (DTU), functional- and pathway enrichment analyses. We show that both magnitude and composition of the responses depend strongly on the combination of host cultivar and pathogen isolate and that DTU appears to be uncommon at this infection stage. We do not detect a set of genes exclusively differentially expressed across incompatible interactions, but genes coding for proteins involved in several defense-related processes consistently show higher expression levels in incompatible than compatible interactions, including receptors and ROS homeostatic, antifungal hydrolytic, phenylpropanoid and diterpenoid biosynthetic enzymes. Our results suggest that whilst interaction-specific differences in magnitude and composition may also shape resistance, different major STB resistances may quantitatively enhance overlapping defense machinery, providing a basis for its functional investigation.

Optical mapping is a fundamental tool for cardiac electrophysiology research. Processing of these signals can be non-trivial, requiring multiple pre- and post-processing steps to limit interpretation errors. Existing software packages have features siloed across platforms and require users to combine several packages for in-depth analysis. To address this, we developed CADENCE, an open-source MATLAB-based application combining feature breadth (e.g., SVD denoising, conduction velocity, alternans, annotated rotors) and depth (e.g., 15 independent alternans features across both voltage and calcium modalities) in an end-to-end pipeline including data conversion, signal conditioning, feature extraction, and signal analysis. We demonstrate CADENCE's capabilities across datasets spanning four species (rat, rabbit, pig, and human), three tissue preparations (slices, wedge, and whole heart), and three camera configurations (single, dual, and panoramic). CADENCE was validated against three blinded expert observers (15 hearts) and demonstrated agreement comparable to inter-observer variability (SD ratios 1.07/0.95/0.89 for APD80, activation, CaTD80). Dominant frequency was recovered with a median error of 0.03% across 84 of 88 paced recordings with 1:1 capture. CADENCE's code is publicly available and distributed as a compiled standalone application that the end-user can install and run without a MATLAB license to encourage adoption and standardization across laboratories.

In many flowering plants, self-incompatibility (SI) is a genetically controlled recognition system that prevents self-fertilization, thereby promoting outcrossing and maintaining genetic diversity within populations. The gametophytic SI system in grasses (Poaceae) is governed by two multiallelic loci, S and Z, each locus comprising of three genes governing the initial recognition between pollen and stigma. Identifying these genes establishes the molecular basis of SI in grasses, but how its unique two-locus architecture has shaped the evolution of the underlying loci and recognition genes remains largely unexplored. Here, we report targeted sequence capture of SI genes in 362 genotypes across nine forage grass species, revealing extensive allelic richness and sequence diversity. The SI genes exhibited elevated nucleotide diversity ({pi}) (0.134 < {pi} < 0.403) and great amino acid sequence divergence, ranging between 48.2-58.0% for the female determinants (sS and sZ) and 66.5-83.4% for the male determinants (SI-DUF247). The SI-DUF247 proteins also displayed trans-generic polymorphism, indicating that alleles predate speciation. Phylogenetic analysis of SI determinants showed congruence at both S- and Z-locus, consistent with co-evolution of recognition partners. Our results demonstrate that the unique two-locus SI system in grasses shares evolutionary characteristics with other SI systems, suggesting that their reproductive function is shaping their evolution. These findings also indicate that the grass SI loci are under long-term balancing selection to maintain recognition specificity while creating sufficient allelic diversity. Our study provides comprehensive sequence resources for SI genes and new insights into the evolution of the unique two-locus SI system of grasses.

Gene regulatory networks (GRNs) establish the spatio-temporal gene expression patterns necessary for reproducible development of organisms. MicroRNAs have emerged as important regulators of gene expression controlling either target gene levels, spatial patterns or timing. These molecular functions may vary depending on how they are integrated into GRNs. Here we used the Arabidopsis leaf as a model to study the genetic and molecular interactions between the CUP-SHAPED COTYLEDON 2 (CUC2) transcription factor and its negative regulator, miR164A. Quantitative morphometric analyses showed that miR164A acts from early stages during leaf development regulating CUC2 expression dynamic rather than the spatial distribution of CUC2. At the molecular level, CUC2 directly activates MIR164A expression, forming a single negative feedback loop. Further, using a ratiometric approach, we identified a cis-regulatory element (CRE) sufficient to drive CUC2-dependent MIR164A expression at the leaf sinus. Ordinary differential equation (ODE) modelling suggests that CUC2-CRE may control the timely accumulation of CUC2. Accordingly, quantification of CUC2 protein accumulation dynamics using a spatio-temporal mean cartography strategy in CRISPR cis-regulatory variants at the MIR164A locus revealed that CUC2-dependent MIRA164A expression is required to maintain both the precise timing of CUC2 expression and CUC2 levels. Altogether, our work provides experimental evidence for a specific role of the CUC2-MIR164A module in shaping leaf development, highlighting the contribution of network topology to GRN functions.

Sudden Death Syndrome (SDS), caused by Fusarium virguliforme, can significantly impact soybean yields. To advance the understanding of SDS resistance in plant introductions, we conducted genome-wide association studies (GWAS) on a set of 351 accessions from the USDA Soybean Germplasm Collection. The analysis revealed seven single nucleotide polymorphisms (SNPs) significantly associated with foliar SDS resistance; three significant QTL (on chromosomes 3, 9 and 11) were detected with Fixed and Random Model Circulating Probability Unification (FarmCPU) and four (on chromosomes 5, 6 and two on chromosome 7) with Bayesian-information and Linkage disequilibrium Iterative Nested Keyway (BLINK). Localized linkage disequilibrium blocks were estimated for these regions. Notably, all of them overlapped with regions where QTL associated with SDS resistance had been previously reported and where within, or close to, sequences annotated as implicated in plant defense mechanisms, such as toxin transporters and transcription factors. The architecture of SDS resistance appears quantitative and polygenic, with most loci explaining modest phenotypic variance. Six plant introductions in this study--PI 175188, PI 307889A, PI 323559, PI 578307C, PI 445683, and PI 323558--harbor favorable alleles at all seven loci, providing high-value resources for resistance introgression. This study underscores the value of leveraging the USDA Soybean Germplasm Collection for the discovery of resistance loci and provides a foundation for marker-assisted breeding to enhance SDS resistance in elite soybean lines.

We performed a screening to evaluate the responses to water deficit before the flowering stage in a panel of 31 genotypes composed of one commercial hybrid, one wild species, multiple ex-PVP (vintage) lines, several commercial heirlooms from the Northeast USA, Mexican landraces from hot and humid environments and genome edited lines with altered flowering and inflorescence traits. We characterized 18 different morphological, physiological and biochemical traits to assess the response to 40% water deficit. We observed diverse morphological, physiological and biochemical responses across the 31 genotypes screened, suggesting the presence of useful variation for breeding for drought tolerance in cultivated tomato. Interestingly, some heirlooms exhibited superior performance under drought, suggesting they can be used as donors to discover novel genes involved in drought responses. Unexpectedly, we found mutants in SlCLE9, a paralog of the stem cell regulator SlCLV3 known to co-regulate meristem proliferation in tomato, exhibited enhanced drought tolerance. Notably, the slcle9 mutant also conferred drought tolerance when used as rootstock with grafted scions from our reference cultivar M82, suggesting SlCLE9 influence root architecture and drought response. Consistent with this, we found SlCLE9 is transcriptionally upregulated in roots and leaves during drought stress, and exogenous application of SlCLE9 modifies root growth and response to drought. Altogether, our screening uncovered new potential trait donors for breeding against drought stress and describes a novel role for SlCLE9 in drought tolerance.

High temperatures during grain filling stage are becoming increasingly frequent, compromising both grain and eating quality and thereby driving demand for heat-resilient cultivars. Such conditions are known to reduce the expression of granule-bound starch synthase I (GBSSI) and starch branching enzyme IIb (BEIIb), which are involved in starch biosynthesis, resulting in a decrease in amylose content and an increase in long-chain amylopectin. Thus, the present study introduced functional mutation in starch synthase IIa (SSIIa) that increases the proportion of short amylopectin chains to genetically compensate for the high-temperature-induced increase in amylopectin long chain. Rice lines carrying the ss2a mutation were grown at two locations with cooler (Akita) and warmer (Okayama) temperatures. Their grain traits, starch structure, and eating quality were compared. The ss2a mutant lines showed an increased proportion of short amylopectin chains as well as an increased apparent amylose content. Furthermore, these alterations in starch structure varied with the grain-filling temperature of the cultivation sites, ultimately affected eating quality. These results suggest that enriching short amylopectin chain via the ss2a mutation can counteract the increase in long amylopectin chain caused by high temperatures during grain filling, thereby maintaining a desirable starch structure and eating quality.

While the core transcription factor TAF4b is known to regulate meiotic transcription and recombination in model plant Arabidopsis thaliana, its functional specialization and evolutionary dynamics remain poorly understood in paleopolyploid crops. To address this, we investigated how the unique, single-copy TAF4b gene modulates early meiotic progression and reproductive success in Brassica rapa. We characterized multiple independent taf4b TILLING lines for fertility and meiotic behavior, then used this information to select a mutant line for further investigation using a pipeline combining whole-genome background sequencing (DNA-seq), stage-specific anther transcriptomics (RNA-seq), yeast two-hybrid assays, and quantitative cytogenetics. Disruption of the meiosis-enriched TAF4b gene on chromosome A7 universally reduced crossover frequency and depleted HEI10 foci, triggering occasional univalent formation. Global RNA-seq and structural modeling indicate that this phenotype is driven by a precise transcriptional dysregulation and altered molecular interactions restricted to the interphase/leptotene transition. In conclusion, TAF4b acts as a stage-specific upstream gatekeeper of the early prophase I expression program in B. rapa, thereby playing an essential, non-redundant role in its meiotic stability.

The mechanisation of agriculture has achieved major increases in labour productivity but has also led to an erosion of agrobiodiversity. Neglected and underutilized crops such as common buckwheat (Fagopyrum esculentum Moench) offer the potential to diversify food production systems but require major genetic improvement to compete with established crops. Although target traits for breeding such as more homogeneous ripening, reduced height, larger seeds and higher and more stable yields are well known, large-scale and field-based phenotyping data of diverse common buckwheat accessions across multiple environments are still scarce. Here, we tested the agronomic performance of a common buckwheat germplasm collection consisting of 170 accessions of diverse origin. The collection was tested at seven year-sites, and 19 agronomic traits related to phenology, growth habit and yield were assessed. Most traits showed high repeatability (0.31 < R2 < 0.98) and heritability (0.45 < H2 < 0.97), indicating genetic control of these traits. Coefficients of variation ranged between 6.19% and 46.15%, showing the high phenotypic diversity within the collection. By combining multiple traits, distinct ideotypes for different roles in the crop rotation were investigated, and accessions with promising agronomic profiles identified. Genome-wide association studies revealed marker-trait associations for 18 traits, allowing the identification of putative candidate genes influencing these traits such as a putative GRF-INTERACTING FACTOR associated with flowering time and biomass production. Our results show that the collection offers vast phenotypic diversity that can be exploited for the development of improved varieties tailored to meet specific agronomic purposes. The efficient utilization of these genetic resources should be supported by the application of genomics-based breeding approaches. The development of genetic markers and search for putative candidate genes mark important steps towards the establishment of efficient breeding schemes. Thus, these genomic resources provide a foundation for accelerating the genetic improvement of common buckwheat and developing agronomically competitive varieties.

Local metabolic programs can determine whether developing organs maintain growth, differentiate, or degenerate. However, the spatial establishment of such programs during reproductive development remains poorly understood. In cereal inflorescences, this question is closely tied to grain number because the fate of initiated floral primordia is shaped by the developmental position, metabolic support, genotype, and environmental stress. To understand the metabolic logic underlying floral fate in barley (Hordeum vulgare L.), we mapped the spatial distribution of metabolites across the inflorescence in wild-type Bowman and in hvcmf4, a mutant that undergoes premature apical inflorescence degeneration. Amino acid-, carbohydrate-, and chlorophyll-associated metabolites form clear developmental gradients along the inflorescence axis. These gradients are progressively remodeled during developmental pre-anthesis tip degeneration but are both accelerated and spatially distorted in hvcmf4. Within this broader metabolic landscape, two discrete domains stand out. First, spermidine-associated domains specifically marked reproductive meristems, a pattern corroborated by the meristem-enriched expression of polyamine biosynthetic genes. This polyamine-rich meristem state declined as developmental tip degeneration proceeded and was prematurely lost in hvcmf4, where polyamine metabolism appeared to shift toward oxidative breakdown via the polyamine oxidase pathway. Second, lysophosphatidylcholine accumulated specifically at floral attachment and vascular supply zones. The developmental reduction of this bioactive lysophospholipid-associated domain in the apical regions, and its stronger disruption in hvcmf4, coincided with the reprogramming of lipid-remodeling and auxin transport-associated genes, suggesting impaired coordination of lipid signaling, auxin canalization, and vascular support. Together, our study reveals spatial polyamine and lysophospholipid domains that couple meristem maintenance with vascular support, providing a metabolic framework for the fate of developing cereal inflorescences.