BMC Plant Biology

Viroids, minimalist plant pathogens, present significant threats to crops by causing severe diseases. The use of high-throughput sequencing technologies for analyzing the transcriptomes of viroid-infected host plants has yielded informative information on gene regulation by these pathogens, however a complete understanding of the transcriptome data suffers from the inclusion of numerous genes of unknown function. Co-expression analysis addresses this by clustering genes into modules based on global gene expression levels. Our previous study emphasized basic helix-loop-helix protein (bHLH) transcriptional reprogramming in tomato in response to different potato spindle tuber viroid (PSTVd) strains. In the current research, we delve into tissue-specific gene modules, particularly in root and leaf tissues, governed by bHLH transcription factors during PSTVd infections. Utilizing public datasets that span Control (C; (mock-inoculated), PSTVd-mild (M), and PSTVd-severe (S23) strains in time-course infections, we uncovered differentially expressed gene modules. These modules were functionally characterized, identifying essential hub genes. We identified the roles of bHLH transcription factors (TFs) in managing processes like photosynthesis and rapid membrane repair in infected roots. In leaves, external layer alterations influenced photosynthesis, linking bHLH TFs to distinct metabolic functions. Expanding on these findings, we explored bipartite networks, discerning both common and unique bHLH TF regulatory roles, notably highlighting the bifan motifs significance in these interactions. Through this holistic approach, we deepen our understanding of viroid-host interactions and the intricate regulatory mechanisms underpinning them.

To address the growing incidences of increased diabetes and to meet the daily protein requirements, we developed low glycemic index (GI) rice varieties with protein yield exceeding 14%. In the development of recombinant inbred lines using Samba Mahsuri and IR36 amylose extender as parental lines, we identified quantitative trait loci (QTLs) and genes associated with low GI, high amylose content (AC), and high protein content (PC). By integrating genetic techniques with classification models, this comprehensive approach identified candidate genes on chromosome 2 (qGI2.1/qAC2.1 spanning the region from 18.62Mb to 19.95Mb), exerting influence on low GI and high amylose. Notably, the phenotypic variant with high value was associated with the recessive allele of the starch branching enzyme 2b (sbeIIb). The genome-edited sbeIIb line confirmed low GI phenotype in milled rice grains. Further, combinations of alleles from the highly significant SNPs from the targeted associations and epistatically interacting genes showed ultra-low GI phenotypes with high amylose and high protein. Metabolomics analysis of rice with varying AC, PC, and GI revealed that the superior lines of high AC and PC, and low GI were preferentially enriched in glycolytic and amino acid metabolism, whereas the inferior lines of low AC and PC and high GI were enriched with fatty acid metabolism. The high amylose high protein RIL (HAHP_101) was enriched in essential amino acids like lysine. Such lines may be highly relevant for food product development to address diabetes and malnutrition. Significance StatementThe increasing global incidence of diabetes calls for the development of diabetic friendly healthier rice. In this study, we developed recombinant inbred rice lines with milled rice exhibiting ultra-low to low glycemic index and high protein content from the cross between Samba Mahsuri and IR36 amylose extender. We performed comprehensive genomics and metabolomics complemented with modeling analyses emphasizing the importance of OsSbeIIb along with additional candidate genes whose variations allowed us to produce target rice lines with lower glycemic index and high protein content in a high-yielding background. These lines represent an important breeding resource to address food and nutritional security.

Small interfering RNAs (siRNAs), a subclass of small non-coding RNAs, play crucial roles in regulating seed germination and viability through epigenetic mechanisms like RNA-directed DNA methylation (RdDM). This study presents the first comprehensive investigation of siRNA profiles linked to seed viability and germination in barley (Hordeum vulgare), utilizing a unique set of seeds from a single batch subjected to controlled long-term storage. Some seeds lost viability due to moisture exposure from unsealing, creating a natural experimental model to explore vigor effects. sRNA sequencing revealed 85,728 differentially expressed siRNAs, with distinct patterns between regenerated, high-viability, and low-viability seeds. Notably, trans-acting siRNAs (ta-siRNAs) showed peak abundance at different imbibition times depending on seed quality, suggesting dynamic regulation. Around 46% of siRNAs were 21 nucleotides, and 54% were 22 nucleotides long. Gene Ontology and degradome analyses confirmed siRNA target genes involved in vital biological processes such as cytochrome complex function, root development, cell maturation, and carbohydrate metabolism. Despite RNA degradation in low-viability seeds, siRNAs remained relatively stable, indicating their potential role in maintaining seed metabolic activity during dormancy release and germination initiation. This pioneering research uncovers novel insights into siRNA-mediated control of seed longevity and germination, highlighting the innovative use of stable, well-characterized plant material to disentangle molecular mechanisms underpinning seed vigor and germination success.

Maize is the most-produced cereal in the world, but its production faces constraints such as parasitic attacks from stemborers. We evaluated the resistance of a core-collection of 18 maize lines by measuring their palatability to European Corn Borer (ECB) larvae fed on maize leaf discs. Using an original consumption test device that takes into account the variability of larvae behaviour, we were able to phenotype the resistance of the 18 maize lines. We matched consumption data to existing enzymatic and metabolomic data that characterized the maize core-collection and identified some metabolites such as caffeoyl-lquinate, trocopherol, digalactosylglycerol and tyrosine that are positively or negatively correlated with the palatability to ECB larvae. Altogether, our results confirm the metabolic complexity involved in the establishment of plant defenses. Metabolic changes associated to leaf palatability mostly concern membrane and cell wall composition. Some of them, pointing-out to the phenylpropanoids pathway, were observed independently of plant developmental pace and plant earliness.

BACKGROUNDGrafting is a horticultural practice used widely across woody perennial crop species to fuse together the root and shoot system of two distinct genotypes, the rootstock and the scion, combining beneficial traits from both. In grapevine, grafting is used in nearly 80% of all commercial vines to optimize fruit quality, regulate vine vigor, and enhance biotic and abiotic stress-tolerance. Rootstocks have been shown to modulate elemental composition, metabolomic profiles, and the shape of leaves in the scion, among other traits. However, it is currently unclear how rootstock genotypes influence shoot system gene expression as previous work has reported complex and often contradictory findings. RESULTSIn the present study, we examine the influence of grafting on scion gene expression in leaves and reproductive tissues of grapevines growing under field conditions for three years. We show that the influence from the rootstock genotype is highly tissue and time dependent, manifesting only in leaves, primarily during a single year of our three-year study. Further, the degree of rootstock influence on scion gene expression is driven by interactions with the local environment. CONCLUSIONSOur results demonstrate that the role of rootstock genotype in modulating scion gene expression is not a consistent, unchanging effect, but rather an effect that varies over time in relation to local environmental conditions.

BackgroundApplication of the wheat phytohormones abscisic acid (ABA) or gibberellic acid (GA) affect Fusarium head blight (FHB) disease severity; however, the molecular underpinnings of the elicited phenotypes remain unclear. Herein, the transcriptomic responses of an FHB-susceptible wheat cultivar Fielder were characterized upon treatment with ABA, an ABA receptor antagonist (AS6), or GA in the presence or absence of Fusarium graminearum (Fg) challenge. ResultsA total of 30,876 differentially expressed genes (DEGs) where identified in Fielder (26,004) and Fg (4,872). Fg challenge alone resulted in the most substantial wheat DEGs contributing to 57.2% of the total transcriptomic variation. Using a combination of topology overlap and correlation analyses, 9,689 Fg-related wheat DEGs were defined. Further enrichment analysis of the top 1% networked wheat DEGs identified critical expression changes within defense responses, cell structural metabolism, molecular transport, and membrane/lipid metabolism. Fg-challenged conditions also included the expression of a putative Fg ABA-biosynthetic cytochrome P450 and repression of wheat FUS3 for dysregulating ABA and GA crosstalk. ABA treatment alone elicited 4536 (32%) wheat DEGs common to those of the Fg-challenge, and Fg+ABA further enhanced 888 (12.5%) of them. These ABA elicited DEGs are involved in defense through both classical and non-classical phytohormone signaling and regulating cell wall structures including polyphenolic metabolism. Conversely, Fg+GA opposed 2239 (33%) Fg-elicited wheat DEGs, including modulating primary and secondary metabolism, defense responses, and flowering genes. ABA and jointly ABAFg[Fg+ABA] treatments repressed, while Fg+GA induced an over-representation of wheat DEGs mapping to chromosome 6BL. Finally, compared to Fg+ABA, co-application of Fg+AS6 did not antagonize ABA biosynthesis or signal but rather elicited antagonistic Fg (557) and wheat (11) DEGs responses directly tied to stress responses, phytohormone transport, and FHB. ConclusionsComparative transcriptomics highlight the effects of wheat phytohormones on individual pathway and global metabolism simultaneously. Application of ABA may reduce FHB severity through misregulating defense mechanisms and cell wall fortification pathways. GA application may alter primary and secondary metabolism, creating a metabolic shift to ultimately reduce FHB severity. By comparing these findings to those previously reported for four additional plant genotypes, an additive model of the wheat-Fg interaction is proposed.

Abstract/SummaryGlandular trichomes are specialized epidermal structures on Cannabis sativa L. inflorescences that synthesize and store cannabinoids and terpenoids, making them central to the species economic and medicinal value. Although trichome development is a multistage genetic process, its regulatory basis in C. sativa remains poorly understood, particularly with respect to sexual dimorphism and sex plasticity. Here, we combined orthology-based gene discovery with transcriptomic profiling to investigate trichome development across four floral phenotypes: female flowers (FF), male flowers (MF), induced male flowers on XX plants (IMF), and induced female flowers on XY plants (IFF). Using trichome development-related genes from Arabidopsis thaliana, a model for unicellular non-glandular trichome development, and Solanum lycopersicum, a model for multicellular glandular trichomes, we identified and mapped 53 candidate C. sativa trichome development regulator genes (CsTDRGs). The CsTDRG set did not support a simple Arabidopsis- or tomato-like model, but instead included Arabidopsis-like epidermal fate components, including MBW-related regulators and GL2, alongside tomato-like multicellular trichome regulators, including MIXTA-like, HD-ZIP IV, WOX, MTR, GRAS, and hormone-associated candidates. RNA-seq analysis showed that CsTDRG expression was more strongly associated with floral phenotype than chromosomal sex. Genes with preferential expression in pistillate tissues were CsTT8, CsMYC1/GL3, CsGL2, CsYABBY4 and CsMIXTA-like1/MYB106, whereas CsMTR1, CsGRAS9, and CsCKX3 were upregulated in male (MF and IMF) flower. These findings suggest that sexually dimorphic trichome development in C. sativa reflects differential regulation of a shared developmental toolkit that combines conserved epidermal fate components with multicellular and glandular trichome regulatory modules.

BackgroundIn the plant sciences, results of laboratory studies often do not translate well to the field because lab growth conditions are very different from field conditions. To help close this lab-field gap, we developed a new strategy for studying the wiring of plant traits directly in the field, based on molecular profiling and phenotyping of individual plants of the same genetic background grown in the same field. This single-plant omics strategy leverages uncontrolled micro-environmental variation across the field and stochastic variation among the individual plants as information sources, rather than controlled perturbations. Here, we use single-plant omics on winter-type Brassica napus (rapeseed) plants to investigate to what extent rosette-stage gene expression profiles can be linked to the early and late phenotypes of individual field-grown plants. ResultsWe find that rosette leaf gene expression in autumn has substantial predictive power for both autumnal leaf phenotypes and final yield in spring. Many of the top predictor genes are linked to developmental processes known to occur in autumn in winter-type B. napus accessions, such as the juvenile-to-adult and vegetative-to-reproductive phase transitions, indicating that the yield potential of winter-type B. napus is influenced by autumnal development. ConclusionsOur results show that profiling individual plants under uncontrolled field conditions is a valid strategy for identifying genes and processes influencing crop yield in the field.

The transformation of the free nuclear syncytium into cellular endosperm tissue with starch and protein accumulation is a well-established phenomenon, at least in the fruits of cereals of the Triticeae tribe. The present article demonstrates that there is considerable diversity inherent in this type of caryopsis morphogenesis. By examining various taxa (species, varieties, and cultivars) of wheat, oats, and some wild grasses, this research reveals significant deviations in endosperm morphogenesis from the typical state. A new developmental pattern of endosperm was identified, characterized by several distinctive features such as incomplete cellularization of the syncytium and starch accumulation within the acellular endosperm domains and the endosperm cavity. A large number of plastids were observed in the syncytium stage, which served as the basis for the later amyloplast stage. The acellular endosperm domains and the cavity domain exhibited connections at specific discontinuities in the modified aleurone layer surrounding the cavity. The peripheral parts of the caryopsis received fewer assimilates necessary for starch synthesis, which was attributed to their increased distance from the transfer system and a likely reduction in the efficiency of assimilate transport through the apoplast in these areas. The starch cavity volume constituted a few percent of the overall caryopsis volume, which could serve as a foundation for potential breeding improvements to enhance starch yields across different varieties.

Dietary fiber composition is a major determinant of fruit nutritional quality, yet its genetic basis remains poorly characterized in wild Vaccinium species. Here, we combined extensive phenotyping with a genome-wide association study (GWAS) to dissect the genetic control of dietary fiber traits in Colombian agraz (Vaccinium meridionale Swartz). Total dietary fiber (TDF), insoluble dietary fiber (IDF), and soluble dietary fiber (SDF), the SDF/IDF ratio, and maturity index (MI) were quantified in fruits from 119 genotypes, representing the most comprehensive evaluation of dietary fiber fractions in fresh Vaccinium fruit to date. GWAS mapped this phenotypic diversity to 24 QTLs distributed across 15 chromosomes, revealing a polygenic architecture underlying fiber-related trait. A TDF QTL (Chr41:26883013) directly co-localized with VaccDscaff31-augustus-gene-268.33, a 7-deoxyloganetin glucosyltransferase, embedded within a glycosyltransferase-rich LD block. IDF variation was associated with VaccDscaff33-processed-gene-116.2 (pectin methylesterase 15) while the SDF/IDF ratio co-localized with VaccDscaff55-augustus-gene-9.30, encoding a xyloglucan endotransglucosylase/hydrolase. Together, the integration of high-resolution phenotyping with QTL mapping connects natural variation in dietary fiber content and composition to specific biosynthetic, remodeling, and regulatory pathways, providing actionable molecular targets for marker-assisted and genomic selection aimed at improving nutritional quality, texture, and processing traits in Vaccinium breeding programs.

Improving cacao yield, a key objective in post-domestication crop improvement, remains a primary goal for breeders, but progress is often hindered by the confounding effects of population structure. To overcome this, we analyzed 346 diverse cacao accessions using an ML-based association mapping framework (with and without population structure adjustment) and a phenotype-only ML prediction of yield. By correcting for population structure, our Bootstrap Forest-based GWAS revealed association signals that showed consistent enrichment for ribosome and protein-synthesis functions, and a recurrent subset of SNPs with high importance appeared across multiple yield components, including pod index and seed number. In parallel, a Neural Network model was utilized to identify cotyledon mass and length as the most powerful predictors for total wet bean mass (R{superscript 2} = 0.715 by repeated five-fold cross-validation), suggesting a practical, low-cost screening proxy for breeding). Collectively, this study delivers a robust genetic framework and a novel predictive tool to accelerate the development of high-yielding cacao varieties through the early identification of elite clones.

Grape breeding programs are mostly focused on developing new varieties with high production volume, sugar contents, and phenolic compound diversity combined with resistance and tolerance to the main pathogens under culture and adverse environmental conditions. The Niagara variety (Vitis labrusca x Vitis vinifera) is one of the most widely produced and commercialized table grapes in Brazil. In this work, we selected three Niagara somatic variants with contrasting berry phenotypes and performed morphological and transcriptomic analyses of their berries. Histological sections of the berries were also performed to understand anatomical and chemical composition differences of the berry skin between the genotypes. An RNA-Seq pipeline was implemented, followed by global coexpression network modeling. Niagara Steck, an intensified russet mutant with the most extreme phenotype, showed the largest difference in expression and showed selection of coexpressed network modules involved in the development of its russet-like characteristics. Enrichment analysis of differently expressed genes and hub network modules revealed differences in transcription regulation, auxin signaling and cell wall and plasmatic membrane biogenesis. Cutin- and suberin-related genes were also differently expressed, supporting the anatomical differences observed with microscopy.

Berry and cluster size are pivotal determinants of grapevine productivity and consumer preferences and remain major targets in grapevine breeding. However, given their complexity as quantitative traits under polygenic control, a deeper understanding of their genetic determinants is needed. The gene pool of the Brazilian grapevine has made a significant contribution to enhancing grapevine performance in tropical and subtropical regions. In this study, we conducted a genome-wide association study (GWAS) using a diverse panel of 288 Vitis spp. accessions from the Instituto Agronomico Germplasm Bank, Brazil. This panel was phenotyped for six cluster architecture traits over 12 years and genotyped using the Vitis18kSNP array. Using two different algorithms, the GWAS identified 56 significant SNPs distributed across 17 chromosomes, validating previously identified quantitative trait loci (QTLs) and revealing novel associations. Four closely spaced markers on Chr1 suggest the presence of a QTL influencing five traits simultaneously. A strong association signal, with phenotypic variance explained (PVE) values of approximately 29-35%, indicated a major QTL for berry length (BL) and width (BWi) on Chr14. Additionally, major-effect SNP loci were identified for cluster weight (CW) on Chr1, cluster length (CL) on Chr7 and 14, cluster width (CWi) on Chr6 and 18, and berry weight (BW) on Chr4, with PVE values ranging from 18-27%. Furthermore, 80 genes associated with berry traits and 52 genes associated with cluster traits were identified as putative candidate genes in the genomic regions associated with significant SNPs. These candidate genes are involved in the regulation of growth and development, hormone regulation, protein synthesis, stress response, and other physiological processes essential for cell health and functionality. Our results provide valuable insights into the genetic determinants of grape berry size and cluster architecture, offering critical data to support future functional studies and enhance the efficiency of related breeding programs.

Plasmodiophora brassicae (Phytomyxea, Rhizaria) is the etiological agent of clubroot disease, one of the most important diseases of Brassicaceae crops. Alteration of metabolism and hormone homeostasis leads to the formation of tumour-like galls in the roots of affected plants. Host plant energy metabolism, defence and developmental processes are under strong temporal control and the very same processes are affected by clubroot. For the first time, this study uses time-resolved transcriptome analyses to explore how P. brassicae affects Arabidopsis thaliana in the night during intermediate (14 days after inoculation; DAI) and late (21 DAI) infection. Day-night differences in gene expression were more pronounced in younger rather than older plants in our differential gene expression (DGE) analysis. Consequently, intermediate phases of infection showed more day-night differences than later ones. Clustering of differentially expressed genes (DEGs) in functional categories highlighted how some of the typical processes known to be disrupted by clubroot infection are more significantly affected in the night and also uncovered some disrupted exclusively in the night. RNA modification stood out as the most unambiguously upregulated process in infected Arabidopsis roots in the night. Analysis of the interaction between clubroot infection and diel oscillations in gene expression detected modifications in the rhythmicity of central circadian clock components during the infection. We discuss our findings in the context of manipulation of plant defence and metabolism, identifying targets for experimental validation and highlighting potential new lines of investigation of our time-resolved datasets to better understand the interaction between P. brassicae and its host. Significance statementPlasmodiophora brassicae impacts physiological processes under strong temporal control in Brassicaceae hosts: e.g., metabolism, hormone homeostasis and defence. Here, for the first time, we performed a time-resolved transcriptomic exploration of clubroot disease at intermediate and mature stages of infection. We identify a previously unrecognised role for nocturnal manipulation of organellar RNA editing and disruption of rhythmicity in circadian clock components. We provide a dataset enabling further exploration of the impact of clubroot on plant circadian processes.

Cassava is a crucial staple crop for smallholder farmers in tropical Asia and Sub-Saharan Africa. Although high yield remains the top priority for farmers, the significance of nutritional values has increased in cassava breeding programs. A notable negative correlation between provitamin A and starch accumulation poses a significant challenge for breeding efforts. The negative correlation between starch and carotenoid levels in conventional and genetically modified cassava plants implies the absence of a direct genomic connection between the two traits. The competition among various carbon pathways seems to account for this relationship. In this study, we conducted a thorough analysis of 49 African cassava genotypes with varying levels of starch and provitamin A. Our goal was to identify factors contributing to differential starch accumulation. With the carotenoid levels of the varieties considered as a confounding effect on starch production, we found that yellow and white-fleshed storage roots did not differ significantly in most measured components of starch or de novo fatty acid biosynthesis. However, genes and metabolites associated with myo-inositol synthesis and cell wall component production were substantially enriched in high provitamin A genotypes. These results indicate that yellow-fleshed cultivars, in comparison to their white-fleshed counterparts, direct more carbon towards the synthesis of raffinose and cell wall components, a finding that is supported by a significant rise in the starch-free residue to total dry yield ratio in yellow storage roots versus white storage roots. Our findings enhance comprehension of the biosynthesis of starch and carotenoids in the storage roots of cassava.

Chilling stress threatens plant growth and development, particularly affecting membrane fluidity and cellular integrity. Understanding plant membrane responses to chilling stress is important for unraveling the molecular mechanisms of stress tolerance. Whereas core transcriptional responses to chilling stress and stress tolerance are conserved across species, the associated changes in membrane lipids appear to be less conserved, as which lipids are affected by chilling stress varies by species. Here, we investigated changes in gene expression and membrane lipids in response to chilling stress during one diurnal cycle in sorghum (Sorghum bicolor), Urochloa (browntop signal grass, Urochloa fusca) (lipids only), and foxtail millet (Setaria italica), leveraging their evolutionary relatedness and differing levels of chilling-stress tolerance. We show that most chilling-induced lipid changes are conserved across the three species, while we observed distinct, time-specific responses in chilling-tolerant foxtail millet, indicating the presence of a finely orchestrated adaptive mechanism. We detected diurnal rhythmicity in lipid responses to chilling stress in the three grasses, which were also present in Arabidopsis (Arabidopsis thaliana), suggesting the conservation of rhythmic patterns across species and highlighting the importance of accounting for diurnal effects. When integrating lipid datasets with gene expression profiles, we identified potential candidate genes that showed corresponding transcriptional changes in response to chilling stress, providing insights into the differences in regulatory mechanisms between chilling-sensitive sorghum and chilling-tolerant foxtail millet. Significance StatementPlants respond to low-temperature stress in myriad ways. While core transcriptional changes are conserved across species, specific adaptive strategies do exist. However, membrane lipid responses during chilling do not appear to be conserved. Here, we collected samples from control and chilling stress-treated seedlings [PSC4] to assess gene expression and membrane lipids in three panicoid grasses to show that lipid metabolic changes follow a daily rhythm. Lipid changes in chilling-tolerant foxtail millet occurred at specific time points, partly explaining the difficulty in finding conserved chilling-induced lipid changes in previous reports. We identified specific orthologs in sorghum and foxtail millet that showed a correlation between gene expression and lipid metabolic changes; these orthologs may be used as potential target genes for developing chilling-tolerant sorghum varieties.

VRS1-5 genes determine spike row types during the early stages of spike development in barley (Hordeum vulgare), yet their functions for the determination of grain yield during the late stages of spike development are largely unknown. To assess the role of VRS1-5 genes in determining grain yield components, we sequenced VRS1-5 genes from 894 worldwide barley accessions and measured 19 spike morphology traits in four environments. Single nucleotide polymorphism SNP markers and gene marker-based haplotypes for VRS1-5 displayed close associations with spike morphology traits. We further developed a spatiote-temporal transcriptome atlas (255 samples) at 17 stages and five positions along the spike, that linked spike morphology to spikelet development and expression patterns of VRS1-5 genes. Phenotypic measurements demonstrated that mutations in VRS1-5 suppress the initiation of spikelet primordia and, trigger spikelet abortion by increasing cytokinin content and improving sensitivity of spikelet primordia to cytokinin. Our integrated results illustrate how breeding can globally alter spike morphology through diversity at the VRS1-5 genes, which show great potential in increasing barley grain yield.

Wheat produces unbranched inflorescences (spikes) composed of smaller inflorescences (spikelets) as their fundamental building units. The spikelet number per spike (SNS) is a major determinant of grain yield and the gene networks that regulate this trait are the focus of this review. Spikelet development starts with the transition of the shoot apical meristem into an inflorescence meristem (IM) that produces lateral spikelet meristems (SMs). The rate at which SMs are produced and the timing of the IM transition into a terminal spikelet (IM[->]TS) determine the final SNS. These two traits are regulated by genes expressed in the IM (e.g. meristem identity genes), as well as by the amount of FLOWERING LOCUS T1 (florigen) transported from leaves to developing spikes. Spikelet number can also be increased by the production of spikes with supernumerary spikelets (SS) or branch-like structures that resemble small spikes. Mutations that promote a reversion from SM to IM identity can induce the formation of SS or branches. Initial efforts to incorporate these mutations into commercial wheat varieties have faced trade-offs in fertility and grain weight, which will require additional research and breeding efforts. Meanwhile, genes and allele combinations that increase SNS without affecting the number of spikelets per node have been identified and are being deployed in wheat breeding programs. Recent spatial transcriptomics, single-cell analyses, and multi-omics studies of wheat spike development are accelerating the discovery of new genes affecting SNS and enhancing our ability to engineer more productive wheat spikes.

BackgroundA diet rich in cruciferous vegetables is reported to have beneficial health effects, partially mediated by 4-methylsulfinylbutyl glucosinolate, or glucoraphanin, which is predominantly found within broccoli (Brassica oleracea var italica). We describe the downstream effects on transcription and metabolism in broccoli following the introgression of a genetic variant of MYB28 into broccoli from a wild Brassica relative which has previously been associated with enhancement of glucoraphanin. ResultsWhole genome sequencing, RNA expression and metabolite analyses were used to characterise the consequences of the introgression of either one or two copies of a genetic variant of the MYB28 transcription factor into a commercial broccoli genetic background. The introgression of the variant of MYB28 resulted in enhanced expression of genes involved in primary sulphate assimilation, sulphur metabolism and aliphatic glucosinolate biosynthesis, and enhanced accumulation of 4-methylsulphinyl butyl glucosinolate in florets. Other changes in transcription that may be related to non-targeted introgression events are reported. There were no consistent effects upon sulphur metabolites pools, apart from methionine-derived glucosinolates. ConclusionThis study illustrates the downstream effects on transcription and metabolism of the introgression of a genetic variant of MYB28 from a wild species into a commercial broccoli genotype.

Crop wild relatives provide valuable insights into trait diversity and the genetic basis of agronomic traits. In the genus Lactuca, domesticated lettuce (Lactuca sativa) and its wild progenitor, Lactuca serriola, have been extensively studied, yet broader wild species remain underrepresented. Here, we present a phenotypic dataset of 550 Lactuca accessions, including 20 wild relatives, capturing plant morphology, pigmentation, and pathogen resistance traits derived from images and genetic resource collections. To investigate the genetic basis of these traits, we used a jointly processed SNP set for L. sativa and L. serriola, applying an iterative two-step GWAS approach, enabling the dissection of multiple loci per trait. We identified both known and novel QTLs associated with anthocyanin accumulation, leaf morphology, and pathogen resistance in L. sativa and L. serriola. Importantly, we identified L. serriola-specific QTLs undetected in L. sativa, revealing unique genetic architectures underlying anthocyanin biosynthesis and leaf morphology in the wild progenitor. These findings expand the knowledge of Lactuca beyond cultivated varieties, highlighting the potential of wild species for breeding applications. Our dataset and results provide a foundation for further investigations into the evolutionary and agronomic significance of Lactuca diversity.