Plant Molecular Biology

G-protein coupled receptors (GPCRs) are responsible for translating environmental signals of various types into cellular signals. Over 40 thousand GPCR orthologs have been discovered in the supergroup Unikonta, and around 800 genes encode for GPCRs in the human genome. In contrast to this astonishing variety, only a handful of GPCR-related genes have been reported in vascular plants, a major group within land plants. In an attempt to advance our understanding of plant GPCRs as well as their role in plant cellular signaling, here we present comprehensive bioinformatic analysis that includes phylogenetic hypotheses, in silico structural analysis, and tissue distribution of transcripts. Altogether, our work strongly suggests that GCR1 is the sole genuine GPCR expressed in Embriophyta. Finally, we briefly discuss the potential role of GCR1 in root hairs, the tubular outgrowths in root epidermal cells that are involved in nutrient absorption, environmental interaction, and root development.

Photoperiod sensitivity (PS) is a key biological response in plants as they adapt to specific environments. Rice (Oryza sativa L.) exhibits a clear PS, as it implements critical phase transition decisions based on PS signals. In this study, we identified a novel PS gene, JMJ706, that is expected to deliver photoperiod-related signals to the flowering-time regulatory network in a day-length-dependent manner. The JMJ706 mutants exhibit early flowering under LD and later flowering under SD compared to WT plants. The gene encodes an H3K9me2 demethylase, and under long-day (LD) conditions, its demethylase activity facilitates the expression of Grain number, Plant height, and Heading-date7 (Ghd7). Since Ghd7 is a floral repressor in LD, it promotes the vegetative phase by delaying flowering. Under short-day conditions (SD), H3K9me2 demethylase activity facilitates Early heading-date 1 (Ehd1) expression, and it acts as a floral accelerator by inducing Heading date 3 (Hd3a) and RICE FLOWERING LOCUS T 1 (RFT1). Furthermore, we propose that the daylength-dependent promotion of target genes (Ghd7 and Ehd1) occurs through demethylation of specific promoter regions at a crucial time window. In addition, JMJ706 may play an important role in regulating plant architecture, including plant height. The natural variation in JMJ706 alleles shows high frequencies across major rice subpopulations, suggesting that JMJ706 could play an important role in the geographical distribution and adaptation of rice cultivars. Our results may add a new layer to the rice flowering-time regulatory pathway, supporting regional adaptation and potential for future breeding.

Phosphorus Starvation Tolerance 1 in rice (OsPSTOL1, known as Phosphorus uptake 1, Pup1) is a receptor-like cytoplasmic protein kinase that confers tolerance to phosphorus deficiency. The OsPSTOL1 gene possesses a Ser/Thr kinase and shows high amino-acid sequence similarity with the leaf rust receptor-like kinase (OsLrK10). We hypothesize that the putative wheat TaPSTOL1 and TaLrK10 have a common ancestral origin and that putative TaPSTOL1 diverged recently acquiring new structural modifications and biological functions in the process. In this study, we identified all putative TaPSTOL1 homeologs and examine the evolutionary relationship between TaPSTOL1 and TaLrK10 in Triticum species. Our results indicate that the putative TaPSTOL1 diverged recently without possessing the amino-terminal domain, which is a typical characteristic of TaLrK10. We observed numerous conversions tracts between these two genes and the substitution pattern of randomly selected amino acids indicates that dynamic selection pressures acted on both genes. The putative TaPSTOL1 shows high nucleotide diversity compared to TaLrK10 within Triticum species. Further, a multiple-sequence analysis reveals that the third exon of TaLrK10 appears to have been duplicated and diverged as a putative single-exon based TaPSTOL1 in bread wheat. Overall, our comparative analysis indicates that both TaPSTOL1 and TaLrK10 appears to have diverged from a common ancestor, acquiring distinct structural organizations and biological functions.

Chloroplast genomes are invaluable resources for plant genomic research, providing insights into genome evolution and molecular adaptation. With the growing scientific and economic interest in Adansonia digitata, a comprehensive characterization of its chloroplast is timely and necessary. A complete chloroplast genome of A. digitata was assembled, annotated, and characterized. Comparative structural analysis was conducted against other Adansonia species, and the assembly was validated through phylogenetic placement within Malvaceae. The assembled genome exhibits the canonical quadripartite organization, spanning 160,061 bp with a GC content of 36.88%, 79 protein-coding genes, 32 tRNAs, and 4 rRNAs. Repeat analysis identified 100 simple sequence repeat motifs, predominantly A/T-rich mononucleotide types (76%), alongside 50 long sequence repeats dominated by forward (26) and palindromic (17) repeats. Comparative analysis with other Adansonia species revealed conserved genome structure, with minor IR boundary shifts involving the ndhF gene, and ycf1 duplication in A. gregorii and A. grandidieri. Average nucleotide identity exceeded 99% across all Adansonia species, with near-complete similarity (ANI {approx} 99.96%) observed with the putative A. kilima. All predicted RNA editing events were nonsynonymous, dominated by C[->]U conversions (55.02%). Codon usage showed non-random synonymous preferences biased toward A/U-ending codons, driven primarily by mutational pressure with detectable gene-specific translational selection. Nucleotide diversity ({pi}) was higher in intergenic spacers (0.00490 {+/-} 0.00574) than in coding regions (0.00167 {+/-} 0.00199), with the majority of genomic regions showing no sequence variation ({pi} = 0). Substitution patterns indicated pervasive purifying selection, with relatively high but insignificant signals in matK, ycf1, accD, and rpoB. Phylogenomic analyses placed the assembled A. digitata chloroplast genome within the Adansonia lineage, consistent with its established systematic position. This study provides detailed insight into the chloroplast genome of A. digitata, and the findings will contribute towards advancing its genomic research.

Previously, the chitin receptor-interacting protein kinase LIK1 (LysM receptor kinase 1/CERK1-interacting kinase) was shown to play an important role in regulating chitin signaling and plant defense. A limited proteolysis proteomics study revealed several LIK1-derived peptides that showed differential abundance between ATP-treated and mock-treated Arabidopsis samples, suggesting a possible involvement of LIK1 in extracellular ATP (eATP) signaling. To explore this possibility, LIK1 mutants were obtained and examined for their response to ATP. The results showed that mutations in LIK1 significantly reduced the expression of eATP-responsive genes. In addition, LIK1 was found to interact with the eATP receptor P2K1 and to be phosphorylated by it. The LIK1 protein was localized to the plasma membrane and its gene expression appeared to be ubiquitous. Collectively, these findings indicate that LIK1 not only contributes to chitin signaling but also participates in eATP signaling, highlighting its potential role as a shared component in multiple signaling pathways to regulate plant responses to diverse internal and external cues.

The RETINOBLASTOMA-RELATED (RBR) protein in plants functions as a cell-cycle inhibitor, regulating cell numbers in developing organs and establishing cellular quiescence during growth. Although the role of RBR counterparts in animals also involves regulating cell size, this potential function remains unexplored in plants. We investigated transgenic Arabidopsis plants with altered RBR levels and observed corresponding changes in cell size from embryogenesis through organ development. In addition, stomatal meristemoid cells with reduced RBR levels divided beyond the size threshold, whereas elevated RBR levels increased their size. RBR stimulated terminal differentiation in the stomatal lineage by inducing MUTE and CYCLIN D5;1 expression, whereas reduced RBR levels maintained asymmetric divisions through high SPEECHLESS and CYCLIN D3;1 expression. Interestingly, the cell proliferation-dependent phosphorylation of RBR at the conserved 911Ser site positively correlated with RBR protein levels in the transgenic lines and aligned with the effect of RBR on cell size. This study discusses the potential link between RBRs control of cell proliferation and cell size, providing new insights into the coordinated regulation of plant development.

Heavy metal ATPases (HMAs) are important group of transmembrane proteins involved in homeostasis of metal ions in plant systems. In this study, a comprehensive analysis of genome assembly (VC1973A v7.1) resulted in the identification of nine HMA genes (VrHMA) and their corresponding proteins in Mungbean, an agronomically important legume crop known for its nutritional values. VrHMA proteins were also characterized based on their biomolecular features, conserved domains and motifs arrangement, transmembrane helices, pore-line helices, subcellular location and occurrence of signal peptides. Based on sequence homology, nine VrHMAs were clustered into two major substrate-specific groups: VrHMA1, VrHMA5 and VrHMA7 were categorized under the Zn/Co/Cd/Pb ATPase group, whereas the remaining six VrHMAs belong to the Cu/Ag subgroup. Gene structure analysis and promoter scanning revealed the structural divergence and presence of various stress-responsive cis-acting elements, respectively. The expression analysis of VrHMA genes in root and leaf tissues, in response to heavy metal (Zn, Cd and Cu) stress, indicates their role in the uptake, transport and sequestration of metal ions. Interestingly, VrHMA5 showed incremental upregulation in roots in response to all three heavy metal stresses, whereas its expression was only upregulated in the leaf tissues under Zn stress, which indicates its role in vascular transport in V. radiata. In addition, this study provides valuable insights into the functional roles of VrHMA genes and will lay a foundation for future genetic improvement in mung bean aimed at enhanced heavy metal stress tolerance and micronutrient homeostasis.

Plant architecture, the spatial configuration of stems, branches, leaves, and inflorescences underpins essential physiological functions such as light capture, assimilate partitioning, flowering, and ultimately, yield. In cassava (Manihot esculenta), architectural traits such us plant height, branching level, and plant shape are agronomically important yet remain underexploited in breeding. Here, a large-scale analysis was conducted using phenotypic and genomic data from more than 14,000 cassava accessions evaluated across 34 field locations in Nigeria between 2010 and 2021, encompassing the national breeding programs of the National Root Crops Research Institute and the International Institute of Tropical Agriculture. The study aimed to dissect the genetic architecture, environmental stability, and breeding relevance of four key traits: plant full height, height to first branching, the branching level number (BranchlevelNum) and plant shape. Phenotypic analyses across breeding stages revealed consistent variation in plant height, branching height, and branching intensity, reflecting the cumulative effects of selection and evaluation across environments. Broad-sense heritability estimates ranged from 0.41 to 0.72, with BranchlevelNum and Cylindrical shape exhibiting strong genetic control and weak correlations with yield components, indicating their suitability for independent improvement. Genome-wide association analyses identified significant loci associated with BranchlevelNum, including a major region on chromosome 2 and an additional locus on chromosome 13, collectively explaining approximately 11% of the phenotypic variance. Candidate genes within these regions included regulators of meristem activity and hormone-related pathways, supporting a developmental basis for branching variation. Genomic prediction accuracy for BranchlevelNum reached 0.44, comparable to values reported for key agronomic traits in cassava. These results demonstrate that branching-related architectural traits are genetically tractable, largely independent of yield, and amenable to genomic selection. The findings support the integration of BranchlevelNum and plant shape into ideotype-driven breeding frameworks aimed at improving flowering efficiency, canopy structure, and field performance in cassava. Author SummaryCassava is a major food crop, and its plant shape plays an important role in how easily it can be grown, harvested, and improved through breeding. Traits such as plant height, branching, and canopy form affect flowering, seed production, and field management, yet they have received much less attention than yield or disease resistance. In this study, we examined plant architecture using field and genetic data from more than 14,000 cassava plants grown across Nigeria over twelve years. We focused on key traits describing plant height, branching level, and overall plant shape. We found that branching level is strongly controlled by genetics, remains stable across environments, and can be predicted accurately using genomic data. We also identified specific regions of the cassava genome linked to branching behavior. Our findings show that plant architecture can be improved using modern breeding tools without compromising yield. Incorporating branching traits into breeding programs can help develop cassava varieties that flower more reliably and perform better in farmers fields.

Improving rice (Oryza sativa L.) yield requires a balanced enhancement of both sink size and source capacity. While many QTLs for sink size have been identified, only a few are known for source capacity, which is essential for achieving high yield. Here we identified qHP10 as a major QTL for increased photosynthetic rate by using chromosome segment substitution lines derived from a cross between the high-yielding indica cultivar Takanari and the average-yielding japonica cultivar Koshihikari. High-resolution mapping combined with CRISPR/Cas9-induced mutagenesis revealed that the causative gene underlying qHP10 is Mitogen-Activated Protein Kinase 4 (OsMPK4). A near-isogenic line carrying the OsMPK4Takanari allele (NIL-OsMPK4) had a 15-25% higher photosynthetic rate than Koshihikari. NIL-OsMPK4 also had higher stomatal conductance than Koshihikari but similar stomatal pore size and density, indicating that increased stomatal aperture increases photosynthetic rate. This enhancement is likely attributable to the down-regulation of OsMPK4 expression, which increases stomatal conductance and thus promotes CO2 uptake. Our findings demonstrate that OsMPK4 is a promising genetic target for increasing source capacity and, potentially, rice yield through molecular breeding. (175 words)

Phenotypic plasticity is a key mechanism by which plants adjust their traits to environmental changes. These phenotypic adjustments are driven by plastic changes in gene expression regulated by gene regulatory networks. Drought, a major selective force in Mediterranean ecosystems, provides a powerful context to examine how genomic plasticity translates into phenotypic responses. Here, we used Dianthus inoxianus, a drought-tolerant Mediterranean carnation, in order to characterize the phenotypic and transcriptomic plasticity in response to drought stress combining ecophysiological measurements with RNA-seq, gene co-expression and gene regulatory network analyses. Most of the phenotypic traits exhibited low plasticity in response to drought, except water and osmotic potential. At transcriptome level, we identified 57 plastic genes, suggesting that drought tolerance in D. inoxianus relies predominantly on constitutive gene expression. These plastic genes were enriched in processes typically related to drought response, such as cell wall components and abscisic acid (ABA) signaling. Some plastic genes belonged to drought-responsive modules, while others were hubs in different modules acting as inter-modular connectors. Furthermore, the regulatory network revealed that these plastic genes were strongly regulated by multiple stress-responsive transcription factors, and that drought-associated modules were regulated through both ABA-dependent and ABA-independent pathways. In addition, we identified contrasting patterns of canalization and decanalization, with immune and post-transcriptional regulation remaining canalized under drought, whereas photosynthesis and amino acid metabolism became decanalized, potentially releasing cryptic genetic variation. Overall, our results emphasise that drought tolerance in D. inoxianus emerges from a strategy combining preadaptation with targeted plasticity in key molecular pathways.

Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are essential regulators of plant growth, development, and stress adaptation. In this study, we performed a comprehensive genome-wide identification of SNARE genes in cucumber (Cucumis sativus L.), uncovering 51 putative members designated as CsSNAREs. Phylogenetic analysis confirmed that these genes cluster into five major clades: Qa-CsSNARE (14), Qb-CsSNARE (9), Qc-CsSNARE (10), Qb+c-CsSNARE (3), and R-CsSNARE (15). Bioinformatic analysis of their promoter regions, coupled with expression profiling under diverse abiotic stress conditions, highlighted a heightened responsiveness within the Qa-CsSNARE subfamily. To validate this, we selected representative Qa-CsSNARE genes for quantitative real-time PCR analysis under drought and salt stress. Among these, CsSYP121 was notably induced by salt treatment. We subsequently generated transgenic cucumber lines overexpressing CsSYP121 and challenged them with salinity. Phenotypic assessment, combined with measurements of reactive oxygen species (ROS) accumulation and K+/Na+ ratios, demonstrated that CsSYP121 overexpression (OE) confers enhanced salt tolerance and boosts antioxidant capacity. We propose a model wherein CsSYP121 mitigates ROS-induced cellular damage under salt stress, potentially through promoting K+/Na+ homeostasis, thereby improving plant performance under saline conditions. Our findings identify CsSYP121 as a promising candidate gene for breeding salt-tolerant crops.

Published studies have reported species-variance between profiles of Calvin-Benson cycle (CBC) intermediates, not only between C4 species and C3 species, but also within C3 species (Arrivault et al., 2019, Borghi et al. 2019). It was proposed that this variance reflects lineage-dependent changes in the balance between different reactions, or poising, of the CBC. These earlier studies investigated phylogenetically-unrelated C3 species. In the current study, CBC intermediates were profiled in five closely-related species from Solanum sect. lycopersicon subsect. Lycopersicum. The levels of individual CBC intermediates showed many significant differences. In a principal component analysis, whilst three species (Solanum lycopersicum, Solanum cheesmaniae, Solanum neorickii) overlapped, Solanum pimpinellifolium and especially Solanum pennellii grouped separately, and were at opposing ends of the distribution. When combined with published data, whilst the separation between Solanum species was retained, they formed a group that was separated from five other C3 species, as well as two C4 species. It is discussed that the observed variation in CBC metabolites profiles within Solanum, together with their separation from other C3 species, supports the idea that CBC evolution is shaped both by phylogenetic relatedness and lineage-specific adaptation. HighlightVariance of intermediate levels points to poising of the Calvin-Benson cycle varying between closely-related species in the tomato clade Solanum sect. lycopersicon subsect. Lycopersicum

The nucleus is the characteristic organelle for eukaryotic organisms. Unlike the classic textbook view of static two-dimensional nuclei, nuclear shape is dynamic inside the live cell. The alteration or deformed nuclear shape is the hallmark of cancer in animal cells and environmental stress in plants. The nuclear envelope proteins interact with chromatin to regulate gene expression. Unfortunately, we have limited knowledge about the impact of abiotic stress on nuclear shape, movement, and chromatin dynamics. To circumvent this issue, we are utilizing a dual fluorescently tagged marker lines - nuclear envelope protein and chromatin - to perform live cell imaging in the model plant Arabidopsis thaliana root. The live cell imaging was performed in control and salt-stressed conditions. We utilized these captured movies to analyze through open-source image processing software Fiji/ImageJ with the help of the TrackMate plugin. Using this method, we have demonstrated that chromatin velocity is decreased in salt-treated conditions. This method will be widely applied to quantitative live cell imaging of nuclear shape and chromatin dynamics during plant development and environmental stress. SummaryThis process aims to simultaneously record nucleus and chromatin dynamics in Arabidopsis thaliana roots and investigate changes in these dynamics in response to developmental and environmental cues.

European ash dieback caused by the invasive ascomycete species Hymenoscyphus fraxineus poses the most prominent danger to common ash trees (Fraxinus excelsior) in Europe. The disease is widely distributed in Europe and currently no efficient management strategy is available. Host-induced gene silencing and exogenous dsRNA applications have shown great potential for controlling fungal diseases in crop plants. In this study, we reported in silico evidence for the presence of a functional RNA interference pathway in Hymenoscyphus fraxineus. Moreover, we showed that the transgenic expression of a double stranded RNA (dsRNA) leads to inhibition of translation of its target polyketide synthase-like gene, a fungal endogene. We explored whether the dsRNA could be introduced exogenously and demonstrated that H. fraxineus can take up externally applied dsRNA molecules. This study highlights the RNA interference mechanism in H. fraxineus and suggests exoRNA applications as a promising approach to control European ash dieback.

Plant genome sequences provide access to the gene repertoire of a species. This facilitates basic research, biotechnological processes, or horticultural applications. Here, we present the genome sequence of Begonia manicata and unravel the genes underlying the pigmentation of red structures emerging from its leaves and stems. Structural genes of the anthocyanin biosynthesis and corresponding regulatory genes were discovered to be upregulated in these red structures suggesting that the pigmentation is caused by the accumulation of anthocyanins. Our work provides a resource for future studies on pigmentation of Begoniaceae.

Subgenome dominance is a phenomenon observed in many allopolyploids where one parental genome exhibits stronger influence over phenotype than the other parental genomes. This may present as preferential retention of one subgenome through fractionation, replacement via homoeologous exchange, or as subgenome expression bias, where one subgenome is expressed at a higher abundance compared to other subgenomes. Sour cherry (Prunus cerasus) is an allotetraploid fruit tree species resulting from an interspecific cross between extant relatives of ground cherry (P. fruticosa) and sweet cherry (P. avium). Prior comparative genomic analyses suggest that the sour cherry cultivar Montmorency contains three subgenomes. Subgenomes A and A, each present in one copy, are derived from a P. fruticosa-like ancestor, and B, present in two copies, is derived from a P. avium-like ancestor. In this study we investigated the subgenome dynamics of the three subgenomes of sour cherry in four diverse landraces and two cultivars, including Montmorency. We found evidence of 26 homoeologous exchange events and five whole-homoeolog replacements relative to Montmorency in three of the six accessions. We also detected subgenome expression bias favoring the A and A subgenomes over the B subgenome, the magnitude of which differs between accessions and changes over the course of fruit development. Lastly, we show differences in dosage variation and expression bias of four previously-described genes in Montmorency associated with fruit softening, a key trait in this crop. These findings on subgenome dominance offer valuable insights into how this phenomenon may influence traits important for sour cherry breeding.

AO_SCPLOWBSTRACTC_SCPLOWNitrogen fertilizers are essential for crop productivity but cause environmental harm, necessitating the development of cultivars that thrive under limited nitrogen. This study investigates the transcriptomic response to nitrate in Arabidopsis thaliana (a model dicot), Brachypodium distachyon (a model Pooideae), and Hordeum vulgare (barley, a domesticated Pooideae) to identify conserved and species-specific molecular mechanisms. Using RNA-seq after 1.5 and 3 hours of nitrate treatment, we found that core nitrate-responsive biological processes - such as nitrate transport, assimilation, carbon metabolism, and hormone signaling - are largely conserved across species. However, comparative analysis at gene level based on orthology revealed specificities between the species. For instance, rRNA processing was uniquely stimulated in Arabidopsis, while cysteine biosynthesis from serine and gibberellin biosynthesis were specifically regulated in Brachypodium and barley. Orthologs of key nitrate-responsive genes (e.g., NRT, NLP, TCP20) exhibited variable regulation, reflecting potential adaptations linked to domestication or nutrient acquisition strategies. These findings highlight the importance of integrating model and crop species to uncover targets for improving nitrogen use efficiency in cereals. The study provides a pipeline integrating gene ontology and orthology analyses to compare transcriptomic responses between species.

Garcinia binucao (Blanco) Choisy is an indigenous species endemic to the Philippines. Its fruit is traditionally used as a souring agent in local cuisine and has been reported to possess nutritional and medicinal properties. Despite its ethnobotanical significance and promising bioactive properties, the species remains underutilized. To date, no genomic resources have been published for G. binucao, limiting its application in food systems, genetic studies, and conservation programs. This study reports the first complete chloroplast genome of G. binucao from an accession conserved at the Institute of Plant Breeding, University of the Philippines Los Banos. The assembled plastome is circular with a length of 156,570 base pairs (bp). It displays the typical quadripartite structure of most angiosperms, consisting of a large single-copy (LSC) region (85,357 bp), a small single-copy (SSC) region (17,129 bp), and a pair of inverted repeats (IR), each 27,042 bp in size. A total of 128 genes were annotated, including 83 protein-coding genes, 37 transfer RNAs (tRNAs), and eight ribosomal RNAs (rRNAs), consistent with the majority of Garcinia species. Of the protein-coding genes, 45 are involved in photosynthesis, 28 genes for self-replication, five genes with conserved open reading frames, and five genes are associated with other functions. The GC content was 36.2%. Leucine (10.6%) was the most abundant amino acid, with a codon usage bias toward UUA. Additionally, 98 simple sequence repeats (SSRs) were detected, 88.78% consisting of A/T motifs. Phylogenomic analysis based on assembled plastome and publicly available cpDNA sequences of 17 other species in the order Malpighiales revealed that G. indica is the closest relative of G. binucao. These findings provide a framework for future research on the species, including its conservation and potential use as a genetic resource.

RNA interference (RNAi) shows great potential to protect crops against fungal diseases, yet reported protection efficiencies vary greatly, and our understanding of the factors responsible for this variance remains limited. In this meta-analysis, we evaluated 89 studies that compare the efficiency of host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS) in controlling fungal diseases, focusing on biotrophic, hemibiotrophic, and necrotrophic fungi, the use of formulations, and the dsRNA design as explanatory factors for differences between reported efficiency values. Our results indicate that SIGS is slightly more effective, particularly in biotrophs. Surprisingly, SIGS studies using formulations did not outperform those applying naked dsRNA. We also assessed parameters of RNA design. Differences in dsRNA length and the number of constructs, and number of targets showed no consistent significant effect on resistance in either HIGS or SIGS. Interestingly, however, HIGS studies reported significantly higher efficiency when targeting genes closer to the 3 end and SIGS when targeting genes closer to the 5 end. We discuss potential reasons for the reported patterns, such as variability in dsRNA uptake mechanisms, intercellular trafficking and Dicer processing, and conclude that more research is needed to understand the biological mechanisms determining RNAi efficiency for fungal control.

The cassava (Manihot esculenta) genome has two large introgressions from its wild relative M. glaziovii on chromosomes 1 and 4 that originate from historical hybridization efforts. The 10 Mbp chromosome 1 introgression has been increasing in frequency in African breeding populations due to its statistical association with higher dry matter content and root number. However, the region also exhibits suppressed recombination, hindering breeders ability to combine favorable glaziovii alleles with the cultivated esculenta background. Since homozygous introgressed lines are rarely selected for advanced trials, dominance effects have not been well-characterized. To analyze the effects of the introgression with higher resolution, we generated a population of over 5000 seedlings from crosses between heterozygous introgressed parents and screened for recombinants using ten KASP markers tagging glaziovii-specific alleles. An optimized subset of 453 lines was then selected and evaluated over two years for yield and vigor traits. Unlike previous studies, composite interval mapping and mixed linear models showed no significant associations between glaziovii alleles and dry matter content or root number. Small, opposing effects on clonal vigor were observed at different ends of the introgression. The region showed significant segregation distortion and enrichment of putative deleterious alleles. Genome alignment of M. esculenta and M. glaziovii assemblies did not show any major structural variants in the introgression region, suggesting that suppressed recombination is likely driven by sequence-level divergence rather than structural rearrangements. These results indicate that the glaziovii introgression does not directly contribute to dry matter, supporting the need for recombination and purging of the glaziovii introgression to aid cassava improvement. Plain language summaryA large chromosome segment from a wild relative of cassava is an important structural aspect in the cassava genome. Since the chromosome segment tends to be inherited as one block, its effects on cassava traits were not well resolved. Through genetic mapping at higher resolution, we identified that the wild segment impacts early vigor and does not appear to impact dry yield, as was previously thought. While there are no major structural differences between the wild and cultivated chromosome segments, their overall divergence seems to suppress the wild chromosome segment from pairing with the cultivated chromosome segment during reproduction. In the apparent absence of any major benefits from the wild segment, removing it from the breeding population may be beneficial. Core ideasO_LIA set of glaziovii allele-specific markers were designed to track the chromosome 1 introgression haplotype. C_LIO_LISegregation distortion suggests the presence of recessive deleterious or lethal alleles in the introgression. C_LIO_LIIncreased recombination is needed to purge deleterious alleles enriched in introgression region. C_LIO_LIThe glaziovii introgression was associated with slightly lower vigor rating and stem diameter. C_LIO_LIThe effects of the previously-identified glaziovii DM QTL were not detected in this population. C_LI