Biology subjects

Cantu, D.

Publications and source records attributed to Cantu, D..

At least 19 recordsLinked to original sources

bioRxiv2026

Thille, a chromosome-level type B avocado genome, and the structural and allelic variation at candidate flowering-type loci

Avocado (Persea americana Mill.) is heterodichogamous: each flower opens first as female and then as male, and cultivars fall into two complementary flowering types, A and B, determined by the transcription factor SDMYB, whose dominant allele confers type A. Avocado genomes have so far come from type A cultivars. Here we report a chromosome-level genome of Thille, a type B cultivar used in breeding. From 72.2 Gb of PacBio HiFi reads (81x coverage) and Omni-C data, the assembly places 859.8 Mb, 96% of the expected 896 Mb genome, in 12 chromosomes with an NG50 of 84.5 Mb, a consensus quality of QV 69.95, and 97.3% complete BUSCO genes; repeats occupy 64.0% of the assembly, and 35,386 protein-coding genes were predicted. We compared Thille with the type A cultivars Hass and Puebla, a predominantly Guatemalan hybrid, and with a West Indian (WI) accession, a Lowland botanical group variety of unreported flowering type. The assemblies differ by large rearrangements, including an 18.8 Mb inversion on chromosome 12 specific to Hass. Using phased reads and haplotype assemblies, we reconstructed the alleles of candidate flowering genes from a type A versus type B Fst scan. Both WI haplotypes carry the recessive SDMYB allele found in Thille; based on this study and the previous study of SDMYB, we predict the WI accession to be type B. Six genes, including SDMYB, carry an allele shared by all three type A cultivars and absent from Thille and WI. The genome and HiFi reads are available under BioProject PRJNA1142556.

genomics↗

bioRxiv2026

Genetic Mapping Reveals a Major Locus for Boron Exclusion in Wild Vitis

Boron toxicity constrains grapevine (Vitis) production in irrigated arid and semi-arid regions, yet most available rootstocks are sensitive to excessive boron. Although underutilized Vitis species may harbor sources of adaptation, the genetic basis of natural variation in boron exclusion remains poorly understood, limiting the development of boron-tolerant rootstocks. We screened 267 wild Vitis accessions representing 16 species for their ability to limit leaf boron accumulation under excess boron conditions. Leaf boron concentration varied 190-fold among accessions and was highly heritable (entry-mean H^2 = 0.71). Its correlation with stomatal conductance was modest (r = 0.34), indicating that transpiration rate explained only a small proportion of the variation in leaf boron accumulation. Leaf boron was correlated with leaf chloride (r = 0.36), but their largely non-overlapping association profiles supported distinct genetic control. Genome-wide association analysis identified a suggestive association on chromosome 11. Independent quantitative trait locus mapping in a full-sib F1 population (n = 158) derived from Longii-9018 (V. acerifolia) x GRN3 (commercial rootstock) identified qBEx11.1 in the same syntenic region on chromosome 11, which explained 19.5% of the phenotypic variance. Positional candidate genes near the association peak included a pectinesterase and a MATE efflux transporter, suggesting roles for cell wall modification and membrane transport. Leaf spectral reflectance was associated with leaf boron concentration in both populations, particularly at visible and red-edge wavelengths, indicating potential for high-throughput phenotyping. Together, these findings identify qBEx11.1 as a priority locus for boron exclusion and support integrated genetic and spectral approaches for breeding boron-tolerant grapevine rootstocks.

plant biology↗

bioRxiv2026

Sixteen thousand grafts reveal limited scion genetic control of grapevine bench-grafting success

Nurseries produce tens of millions of bench-grafted grapevines (Vitis vinifera L.) each year, yet the genetics underlying grafting success rate remain poorly understood. Here, we dissected the genetic architecture of this trait in a scion mapping population. We grafted 138 progeny from a Riesling x Cabernet Sauvignon cross, along with both parents, onto 1103P, with 40 grafts per genotype in each of three separate field blocks and grafted on separate days (16,645 grafts). Success averaged 91.1 %. Scion genotype explained 19 % of the variance among blocks (entry-mean heritability 0.42); genotype x block variance exceeded genotype variance, and a few genotypes failed almost completely in a single block. Success was uncorrelated with yield of the mother vines. QTL mapping across six phenotype definitions and four haplotype-resolved parental genome assemblies detected one modest, reproducible QTL on chromosome 7 (Cabernet Sauvignon alleles; +3.9 percentage points; 11 to 15 % of variance) and suggestive regions on chromosomes 5 and 9. Most variation in grafting success within this elite V. vinifera cross was non-genetic and attributable to events affecting entire bundles. These results set expectations for effect sizes, replication, and process control in genetic studies of grafting success.

genetics↗

bioRxiv2026

Harnessing Vitis germplasm diversity to dissect and predict adventitious rooting traits in grapevine

Adventitious root formation (ARF) is a critical trait for the cost-effective propagation of grapevines in commercial nurseries. Poor rooting ability can limit the use and adoption of new rootstocks derived from underutilized Vitis species, constraining breeding efforts largely to the traditional trio: Vitis riparia, V. rupestris, and V. berlandieri. Despite its agronomic relevance, the genetic basis of ARF remains poorly characterized across the broader Vitis genus. In this study, we evaluated 308 accessions representing 18 Vitis species over three growing seasons, quantifying rooting performance at two developmental stages, callus-stage and post-transplant, alongside root biomass, cutting weight, and a derived transplant-response index. We observed extensive phenotypic variation both within and across species, and species rankings depended on the trait considered. V. riparia, V. rupestris and V. californica ranked among the top five species for all four rooting traits, whereas V. cinerea and V. candicans ranked among the lowest for root weight and post-transplant rooting. V. arizonica and V. acerifolia rooted well at the callus stage but were intermediate after transplanting, and V. berlandieri was among the weakest at the callus stage yet intermediate for post-transplant rooting. Repeatability was moderate to high for root weight (0.74) and callus-stage rooting (0.66), and lower for post-transplant rooting (0.47), reflecting both genetic control and season-to-season variation. Between-species differences accounted for 68% of the genetic variance in callus-stage rooting but only 10% in cutting weight. Rooting was associated with the climate of each accession's wild site of origin: after removing differences among species, accessions originating from sites with lower dry-season precipitation rooted better and produced more root biomass. Genome-wide association analysis using 3.4 million SNPs identified 54 significant SNPs resolving into 18 independent loci across four traits, with root weight contributing 12 of them. Candidate genes in linkage with these loci include a mitogen-activated protein kinase, a SCARECROW-LIKE GRAS transcription factor, PASTICCINO1, expansin A1, an AP2/ERF-RAV1 transcription factor, a tandem array of caffeoyl-CoA O-methyltransferases, and several sugar, peptide and nitrate transporters, implicating auxin-linked cell proliferation, cell wall and lignin remodeling, and solute transport. Genomic and phenomic prediction models yielded moderate accuracies across traits and seasons; up to r = 0.67 for post-transplant rooting within a season and r = 0.65 for previously unevaluated accessions. Moreover, the integration of spectral and genotypic data further improved predictive performance. Prediction accuracy was essentially flat between 5,000 and 50,000 markers. This study establishes a foundational framework for the genetic improvement of grapevine rootstocks, promoting broader use of resilient, high-performing, and clonally-propagable germplasm in viticulture.

genetics↗

bioRxiv2026

TPS Expression, Not Copy Number, Explains Aroma Diversity among Grapevine (Vitis vinifera L.) Cultivars

Terpenoids are major contributors to grapevine berry and wine aroma, with mono- and sesquiterpenoids exhibiting substantial variation among cultivars. However, the genetic basis of cultivar-specific terpenoid profiles remains only partially understood due to gene duplications that expanded the terpenoid synthase (TPS) gene family across different cultivars. Here, we compared haplotype-resolved genome assemblies for seven cultivars to analyze copy number variation and expression profiles of terpenoid biosynthesis genes and terpenoid accumulation throughout flower and berry development. These assemblies resolved duplicated TPS loci at the haplotype level, revealing extensive copy number variation across cultivars and between haplotypes within a cultivar, with up to 44 mono-TPS genes on a single haplotype. We identified eight specific monoterpenoids as key drivers of cultivar-specific profiles in ripe berries. Total TPS transcript abundance correlated strongly with total terpenoid accumulation over time. In contrast, TPS copy number was largely decoupled from both expression and terpenoid accumulation, indicating that expression regulation, rather than gene dosage, underlies aroma differences among cultivars. Transcript-metabolite network analysis revealed both expected correlations between specific TPS genes and putative products as well as novel associations. We functionally validated selected candidate TPS in Nicotiana benthamiana, characterizing -farnesene, {beta}-ocimene, and multi-product -terpineol synthases. The in vivo characterization demonstrated that minor sequence variations can alter catalytic activity and product profile. Together, these findings show that the extensive duplication of TPS genes in grapevine does not itself determine terpenoid output; instead, cultivar-specific aroma arises from how these genes are expressed and from sequence differences that shape enzyme activity.

plant biology↗

bioRxiv2026

Genomic compartmentalization of pervasive sex-biased gene expression in the vine mealybug Planococcus ficus

The vine mealybug, Planococcus ficus, is a globally invasive pest of grapevine and a vector of leafroll viruses. Like other mealybugs, it reproduces through paternal genome elimination, a sex-determination system that operates without sex chromosomes and is associated with extreme sexual dimorphism. To characterize genome organization and sex-biased expression in this species, we generated a long-read reference genome spanning 369 Mb with 23,489 annotated genes and macrosynteny conserved with the citrus mealybug, Planococcus citri. Resequencing of four California field individuals yielded a first whole-genome estimate of nucleotide diversity and 132 microsatellite markers for population monitoring. Among 2,129 candidate secreted proteins, a conserved core is shared with P. citri, but each species carries a distinct set of lineage-specific effectors. Comparing adult male and female transcriptomes, we found sex-biased expression to be pervasive and skewed toward females: 41% of tested genes differed between the sexes, with female-biased genes both more numerous and showing larger fold changes. These female-biased genes were not randomly distributed but concentrated in discrete blocks of coordinately expressed, tandemly duplicated gene families, a pattern not previously described in a mealybug. Male- and female-biased secreted proteins also differed in origin, with male-biased proteins drawn from a conserved repertoire shared with P. citri and female-biased proteins spanning a more lineage-specific pool. Together, these results reveal a female-skewed, spatially clustered architecture of sex-biased expression in a mealybug that lacks sex chromosomes, and provide genomic resources for managing an invasive vineyard pest.

genomics↗

bioRxiv2026

Botrytis cinerea infection reshapes the grape berry microbiome during noble rot

Noble rot, caused by Botrytis cinerea, profoundly alters grape berry physiology and is essential to produce botrytized wines. In this study, we profiled bacterial and fungal communities associated with Vitis vinifera cv. Semillon berries across four stages of noble rot development and four consecutive vintages using 16S rRNA gene and ITS1 amplicon sequencing. Noble rot stage significantly impacted the structure of bacterial communities across vintages, while fungal communities showed more variable vintage-dependent responses. Bacterial alpha diversity increased consistently at advanced stages of infection (S3), coinciding with a marked shift from Pseudomonas-dominated communities toward acetic acid bacteria, particularly Gluconobacter, which was significantly enriched at S3 in all vintages. Fungal communities remained dominated by Sclerotiniaceae throughout infection, consistent with pervasive B. cinerea colonization, while non-Botrytis fungal taxa shifted from filamentous fungi such as Cladosporium and Alternaria toward fermentative yeasts including Hanseniaspora and Lachancea. Co-occurrence network analysis revealed a positive association between Gluconobacter and these fermentative yeasts, suggesting coordinated enrichment of oxidative and fermentative microorganisms at advanced noble rot stages. Together, these results reveal a reproducible stage-associated microbial succession during noble rot progression and identify acetic acid bacteria as consistent markers of advanced infection.

microbiology↗

bioRxiv2026

Allelic variation at a terpene synthase locus shapes wine monoterpene composition and aroma

The genetic basis of wine monoterpene variation and its sensory consequences remain poorly understood. We investigated how allelic variation at a chromosome 10 linalool QTL harboring a terpene synthase cluster, including VviTPS54, influences wine aroma in a Riesling x Cabernet Sauvignon mapping population. Twenty-six wines from progeny selected for contrasting genotypes were subjected to HS-SPME-GC-MS profiling and descriptive sensory analysis. Genotypes carrying the high-linalool Riesling allele produced substantially elevated monoterpene levels dominated by (3S)-linalool. Berry and wine monoterpene profiles were significantly correlated, and berry monoterpene glycosides showed broad correlations with wine monoterpenes. High-monoterpene genotypes were more closely associated with Floral, Tropical Fruit, and Apricot/Peach attributes, though perception was modulated by matrix effects from berry pigmentation. These results demonstrate that allelic variation at a terpene synthase locus drives coordinated changes in wine monoterpene composition and linked sensory attributes, while underscoring the complexity of translating genetic variation into sensory outcomes.

genetics↗

bioRxiv2026

Cell-type-specific transcriptional plasticity in a multi-organ atlas reveals organ-independent companion cell regulatory programs in grapevine

Plants comprise both cell types shared across organs and those restricted to specific tissues. How the transcriptional programs defining cell identity are maintained or remodeled across organ contexts remains poorly understood, particularly in long-lived perennials, for which cell type-resolved transcriptomic data remain scarce. We generated a multi-organ single-nucleus transcriptomic atlas of the dwarf grapevine cultivar Pixie, comprising over 220,000 nuclei from nine organs, including roots, green stems, pre-anthesis flowers, dormant buds, young and old leaves, and berries at three developmental stages, each sampled in duplicates. We annotated 46 distinct cell types, reconstructed developmental trajectories within selected cell types, and inferred gene regulatory networks at cell type resolution. Broadly distributed cell types, including epidermis, xylem parenchyma, and phloem parenchyma, exhibited pronounced organ-dependent transcriptional divergence, with organ identity accounting for 65% of regulon activity variance across the atlas. In contrast, companion cells maintained organ-independent regulatory programs, representing the stable end of a continuum of transcriptional plasticity that spans shared cell types. We identified cell-type-specific transcription factor expression and inferred gene regulatory networks using motif-based regulon analysis, revealing candidate regulators of cell identity and tissue specialization. Together, this atlas provides a reference framework for cell type-resolved functional genomics in a perennial woody crop.

genomics↗

bioRxiv2026

Haplotype-specific expression of a terpene synthase underlies linalool variation in the grapevine cultivar Riesling

Grapevine cultivars vary widely in monoterpenoid content, yet the genetic and regulatory mechanisms underlying this variation remain poorly characterized beyond highly aromatic Muscat types. We profiled free volatiles and monoterpenoid glycosides in a Riesling x Cabernet Sauvignon F1 mapping population, revealing extensive variation and transgressive segregation consistent with multigenic control. QTL mapping identified 59 significant loci associated with 47 volatile compounds and monoterpene glycosides, including two major QTLs explaining 33.8% and 32.4% of phenotypic variance in (3S)-linalool accumulation. Integration of haplotype-resolved transcriptomics with metabolite data, enabled by a chromosome-scale diploid Riesling genome assembly, resolved a (3S)-linalool/nerolidol synthase cluster on chromosome 10 and identified VviTPS54 as the strongest candidate underlying linalool variation. VviTPS54 exhibited haplotype-specific expression strongly correlated with (3S)-linalool accumulation across genotypes, while no QTL was detected at the VviDXS1 locus and VviDXS1 expression was not correlated with terpene levels, indicating that regulatory variation within terpene synthase clusters, rather than methylerythritol phosphate (MEP) pathway flux, drives monoterpenoid composition in this population. These results establish regulatory variation of terpene synthases as a key mechanism underlying monoterpenoid diversity in grapevine and demonstrate that resolving such variation requires haplotype-phased genome assemblies coupled with haplotype-resolved transcriptomics to detect allele-specific expression differences at complex, heterozygous loci.

plant biology↗

bioRxiv2026

Genome-wide characterization of extant clonal diversity in Chilean Carmenere

Carmenere is a widely cultivated and internationally recognized grapevine cultivar in Chile, yet genetic variation among its clones remains poorly characterized. Early studies based on SSR and AFLP markers detected limited polymorphism, but these approaches interrogate only a small fraction of the genome, leaving the extent of clonal diversity unresolved. Here, we generated an improved chromosome-scale diploid genome assembly of Carmenere FPS clone 02 and characterized clonal genomic diversity by sequencing 36 biological replicates representing 12 clones maintained in Chile, including heritage selections rescued from old producer vineyards by Vina Santa Carolina as part of its Bloque Herencia conservation program, and commercial nursery-derived clones. Focusing on low-frequency variants and using replicate-aware consensus calling, we identified more than 9,000 private single nucleotide variants (SNVs) and small indels per clone, providing high-resolution markers for clonal identification. Although most variants were located in repetitive or intergenic regions, a subset affected coding sequences, with genes involved in plant-pathogen interactions, transport, and secondary metabolism most frequently impacted. While variant-affected genes associated with wine anthocyanin content, TA, pH, and alcohol percentage were identified, broader phenotypic characterization will be required to assess their biological significance. Overall, this study provides a genome-wide characterization of extant clonal diversity in Carmenere, with implications for clonal selection and genetic resource conservation.

genomics↗

bioRxiv2026

The dynamics of introgression across an adaptive radiation: examining hybrid speciation and parallel adaptation in North American Vitis

Hybridization between species can contribute to parallel adaptive events and, in some cases, speciation. However, the prevalence of introgression is rarely measured across multiple species, meaning that it is rarely integrated with landscape-scale processes or connected to adaptive repeatability. Here, we analyzed whole-genome resequencing data from 639 accessions representing 48 Vitis species. Vitis is notable for the domesticated grapevine (V. vinifera), multiple economically important North American species, and as an example of a temperate adaptive radiation. Our dataset included population-level sampling for 19 species, from which we reconstructed individual- and species-level phylogenetic frameworks for the genus. The analyses uncovered widespread evidence of introgression, comprising [~]14% of the average Vitis genome. Introgression was associated with geographic distribution between species, and highly admixed individuals were more frequently found near ecological niche margins. Genetic analyses further indicated that previously recognized hybrid taxa, V. x doaniana and V. x champinii, likely represent hybrid swarms rather than distinct species. We also assessed patterns of adaptive repeatability across eight species with denser population sampling. For six of the eight species, most of the detected sweeps overlapped with sweeps in other species; on average, more recently diverged species shared more overlapping sweeps. Parallel sweep events were driven mostly by introgression, although a substantial fraction ([~]40%) was attributed to selection on ancestral standing variation. By integrating population genomic data across many species, this study highlights the central role of shared genetic variation across an adaptive radiation.

evolutionary biology↗

bioRxiv2025

Haplotype graph analysis of PdR1 uncovers resistance diversity to Pierce's Disease in Vitis arizonica and its hybrids

Previous genetic mapping studies indicate that multiple haplotypes of the Pierces disease (PD) Resistance 1 (PdR1) locus occur in Vitis arizonica and its hybrids. To characterize sequence diversity at this locus, we assembled chromosome-scale diploid genomes for four PD-resistant (PD-R) accessions: b43-17 (PdR1a/PdR1b), the backcross 07744-094 (PdR1c/PdR1-), b46-43 (PdR1e/PdR1f), and b42-26 (PdR1-/PdR1-), which displays quantitative PD resistance not associated with PdR1. Haplotype resolution of PdR1a, PdR1b, PdR1c, and PdR1e revealed substantial variation in intergenic repeat content and gene composition between PdR1 and their alternative haplotype at the PdR1 locus not associated with PD resistance phenotype (PdR1-), as well as among PdR1 haplotypes, demonstrating extensive sequence diversity at the PdR1 locus. Sequence graph analysis uncovered substantial structural divergence concentrated in approximately one quarter of the locus, together with smaller-scale variation across haplotypes. This analysis identified PdR1-specific graph nodes, showing that PdR1a and PdR1b share most of their PdR1-specific features, whereas PdR1c contains the highest number of private nodes, followed by PdR1e. Integration of sequence graph features with gene expression data further refined a set of defense-related candidate genes within PdR1c. Together, these results identify candidate genes for functional validation and indicate that multiple resistance determinants co-localized within the PdR1 locus may contribute to PD resistance, highlighting opportunities for targeted genetic improvement strategies.

genomics↗

bioRxiv2025

Dual single-nucleus gene expression atlas of grapevine and Erysiphe necator during early powdery mildew infection

We applied dual-organism single-nucleus transcriptomics to study the interaction between grapevine leaves and Erysiphe necator, the causal agent of powdery mildew, at one and five days post-infection, including controls and three biological replicates. We generated a grapevine leaf atlas encompassing over 100,000 nuclei, and a pathogen atlas of more than 3,000 nuclei. We successfully annotated all major grapevine cell types, including mesophyll, epidermis, phloem and xylem parenchyma, companion cells, and guard cells. We identified key E. necator structures, including appressoria, haustoria, and hyphae, and provided a list of novel cell type markers for both species. We reveal structure-specific gene expression programs in E. necator laying a foundation for future studies of fungal development and virulence mechanisms. In the host, we identified spatially distinct expression patterns of defense-related genes. As the infection progressed, we observed the activation of a coordinated immune response involving multiple cell types, mainly epidermal and mesophyll cells. High-dimensional weighted gene co-expression network analysis identified key hubs and networks associated with cell type-specific signaling and defense response. We describe a spatial separation of pattern- and effector-triggered immunity, supporting a model in which pattern-triggered immunity is activated at the site of pathogen contact and effector-triggered immunity is induced in surrounding tissue.

plant biology↗

bioRxiv2025

Stress-dependent responses of grapevine wood and fungal pathogen activity under esca and drought

O_LIBiotic and abiotic stresses alter the physiology of perennial plants, with consequences for fungal endophytes and disease expression. In grapevine, one of the worlds most valuable crops, drought inhibits esca disease expression, but the underlying molecular interactions between plant and fungi are unknown. C_LIO_LIWe combined wood metatranscriptomics, metabolomics, and metabarcoding to investigate these interactions in 30-year-old grapevines and eight wood-pathogenic fungi under conditions of drought or esca leaf symptom expression. C_LIO_LIBoth esca and drought decreased grapevine transpiration, but with different transcriptomic and metabolic signatures. Similar pathways were also activated, including the phenylpropanoid and stilbenoid synthesis pathways. These stress responses could potentially confer cross-tolerance, and elicit different fungal molecular responses. The levels of putative fungal virulence factors increased significantly under both stresses. Under drought, only the relative abundance of Phaeomoniella chlamydospora and gene expression involved in anti-oxidative mechanisms, growth, and reproduction increased. Under esca expression conditions, only the relative abundance of Fomitiporia mediterranea and gene expression involved in wood degradation, competition, detoxification, and growth increased. C_LIO_LIThe grapevine defense mechanisms induced by drought coupled with a low transpiration rate and a low abundance and virulence of F. mediterranea may account for esca leaf symptom inhibition upon water deficit. C_LI

plant biology↗

bioRxiv2025

Intraspecies sequence-graph analysis of the Phytophthora theobromicola genome reveals a dynamic structure and variable effector repertoires

Phytophthora theobromicola is an emerging cacao pathogen recently identified in Brazil as an aggressive agent of black pod rot. We generated genome assemblies for two P. theobromicola isolates using long-read sequencing and five additional isolates using short reads. Comparative analysis revealed a genome size and predicted gene content comparable to P. citrophthora, a closely related species with a broad host range that includes both citrus and cacao. An intraspecies sequence-graph analysis revealed a highly dynamic genome structure with high proportion of variable effectors. Syntenic orthology analysis across 13 Phytophthora species identified orthologous gene groups conserved only in cacao pathogens and others specific to P. theobromicola. RxLR effectors and CAZymes were particularly enriched among lineage-specific syntenic groups, with RxLRs preferentially located near transposable elements and within gene-sparse, repeat-rich regions. Transcriptome analysis of infected cacao tissues showed that 88% of predicted effectors were expressed, with pods exhibiting the highest number of upregulated genes. Notably, several RxLRs classified as P. theobromicola-specific syntenic orthologs were highly expressed in infected tissues, suggesting that these lineage-specific effectors may play key roles in host-pathogen interactions unique to cacao. Together, our findings highlight the dynamic architecture and functional plasticity of the P. theobromicola genome, providing foundational insights into its virulence strategies and supporting future studies on host adaptation and effector evolution in emerging cacao pathogens.

genomics↗

bioRxiv2025

Phased epigenomics and methylation inheritance in a historical Vitis vinifera hybrid

Epigenetic modifications, such as DNA methylation, regulate transcription and influence key biological traits. While many efforts were made to understand their stability in annual crops, their long-term persistence in clonally propagated plants remains poorly understood. Grapevine (Vitis vinifera) provides a unique model, with cultivars vegetatively propagated for centuries. Here, we assembled the phased genomes of Cabernet Sauvignon and its parental lineages, Cabernet Franc and Sauvignon Blanc, using HiFi long-reads and a gene map tenfold denser than existing maps. Using three clones per cultivar, we quantifyied methylation with very consistent short- and long-read sequencing and ensured both varietal representativeness and assessment of clonal variability. We leveraged the parent-progeny sequence graph to highlight allele-specific methylation and conserved transcriptomic patterns for genes and small RNA. Such a format was essential to integrate multi-omics data and revealed that despite less clonal conservation than genetic polymorphisms, methylation marks were remarkably inherited. By further demonstrating the linear-reference limitations, we determined the correct representation of genetic variants by the sequence graph is crucial for the accurate allelic quantification of the methylome. These findings reveal the remarkable stability of epigenetic marks in a model propagated by asexual reproduction. Using a phased sequence graph, we introduce a scalable framework that accounts for genomic variation, accurately quantifies allele-specific methylation, and supports multi-omics integration such as our evaluation of the transcriptional impact of epigenetic inheritance. This approach has broad implications for perennial crops, where epigenetic variation could influence traits relevant to breeding, adaptation, and long-term agricultural sustainability.

genomics↗

bioRxiv2025

Integration of diploid T2T parental genomes with progeny genomes and transcriptomes identifies resistance genes in the grape powdery mildew resistance loci Ren6 and Ren7 of Vitis piazeskii

The Chinese grape accession Vitis piasezkii DVIT2027 carries two loci associated with powdery mildew (PM) resistance loci, Ren6 and Ren7, which differ in timing and strength of response to Erysiphe necator. Both loci are consistent with recognition by intracellular immune receptors. To identify the underlying nucleotide-binding leucine-rich repeat (NLR) genes, we assembled chromosome-scale diploid genomes of DVIT2027 and the susceptible V. vinifera F2-35, parents of a segregating F1 population. We integrated these assemblies with deep resequencing data from eight F1 sib-lines carrying different Ren6/Ren7 combinations and generated trio-binned, parent-phased genomes for six progeny. This resolved both PM-resistant (PMR) and PM-susceptible (PMS) haplotypes at Ren6 and Ren7. Comparative analyses revealed extensive structural variation and complete haplotype specificity among NLRs, with several candidate genes lacking allelic counterparts in PMS haplotypes. Expression profiling across PMR sib-lines identified four and two CC-NBS-LRR genes potentially associated with Ren6 and Ren7, respectively. Sequence graph reconstruction of these loci across multiple V. piasezkii accessions revealed broad intraspecific diversity and DVIT2027-specific nodes, including within candidate NLR genes. These results provide a high-resolution view of Ren6 and Ren7 and support the identification of resistance gene candidates for functional validation and grapevine breeding.

plant biology↗