Biology subjects

Michael, T. P.

Publications and source records attributed to Michael, T. P..

At least 19 recordsLinked to original sources

bioRxiv2026

A haplotype-resolved, near telomere-to-telomere genome assembly of the desiccation-tolerant plant Vellozia tubiflora

Vegetative desiccation tolerance allows plants to survive the loss of up to 95% of their cellular water content and recover upon rehydration. The family Velloziaceae contains the largest number of desiccation-tolerant species among flowering plants, yet genomic resources for this lineage remain extremely limited. Here we report the first reference genome of the genus Vellozia, the most species-rich genus of the family. Using PacBio HiFi sequencing and Omni-C chromatin conformation data, we generated a haplotype-resolved genome assembly of Vellozia tubiflora with near telomere-to-telomere continuity. Hap1 and Hap2 span 423.2 Mb and 426.4 Mb, respectively, with both assemblies successfully anchored into nine chromosome-length scaffolds. The former presents a scaffold N50 of 49.8 Mb and 96.5% BUSCO completeness, while the latter reaches 50.0 Mb and 96.8%. Genome annotation predicted 23,352 and 23,549 protein-coding genes in Hap1 and Hap2, respectively. This resource is anticipated to foster new insights for comparative and evolutionary studies of vegetative desiccation tolerance across angiosperms and expands the genomic representation of plants from the Brazilian campos rupestres, a threatened biodiversity hotspot characterized by extreme environmental conditions.

plant biology↗

bioRxiv2026

Inherited proliferation states organize plant transcriptomes

A central question in plant biology is whether natural transcriptome variation primarily reflects environmental adaptation or intrinsic genetic programs. Using transcriptomes from 665 natural Arabidopsis thaliana accessions, we show that inherited population structure, not environmental gradients, is the primary organizer of global transcriptome architecture. Genomic population structure uniquely explains 30.0% of transcriptome-wide variation, while environmental variables account for only 2.5%. This architecture is dominated by a single co-expression program (proliferation Module Eigengene: prolifME), enriched for cell cycle regulation and ribosome biogenesis, regulated by a bipartite transcription factor architecture, and genetically encoded at discrete loci with broad trans-regulatory effects. This proliferation program is associated with reduced plant size, biomass, growth rate and water-use efficiency, and these relationships persist under common-garden conditions. Gene contributions to this proliferation program are non-randomly conserved in rice and maize across 150-200 million years of divergence. These findings identify an intrinsic transcriptional proliferation program as a primary state of plant population transcriptomes, conserved across plant species.

plant biology↗

bioRxiv2026

Deciphering Parasitic Strategies: Dual Transcriptomics Reveal Distinct Infection Mechanisms and Gall-like Traits in Rafflesiaceae

Rafflesiaceae, known for producing the largest flowers in the world, are obligate parasites that exclusively infect Tetrastigma sp. (Vitaceae). Despite their unique biology, the interactions between parasitic tissues and host roots remain poorly understood, particularly during the flower morphogenesis phase, where parasitic tissue erupts through the host root. Here, we performed dual transcriptome analyses of two Rafflesiaceae species and their respective Tetrastigma hosts: Sapria himalayana with T. cauliflorum and Rafflesia speciosa with T. magnum. Our findings reveal species-specific transcriptional responses in Tetrastigma, suggesting divergent parasitism strategies between Rafflesia and Sapria. Moreover, we identify molecular signatures of parasitism that parallel plant gall formation, particularly in genes governing cell wall modification and host tissue reorganization. Unlike bacterial or insect-induced galls, these mechanisms may involve fungal symbionts, highlighting the unique nature of these interactions. Together, our results demonstrate that Rafflesiaceae parasitism represents a complex tripartite relationship among host, holoparasite, and associated microbes, offering new insights into the hidden biology of these remarkable parasitic plants.

plant biology↗

bioRxiv2026

Whole-genome phylogenomics and synteny resolve a single origin of body-plan asymmetry in flatfishes

Flatfishes display the most dramatic asymmetric body plan in vertebrates, yet whether this rare innovation evolved once (flatfish monophyly, FM) or multiple times (flatfish polyphyly, FP) has remained contentious. A recent genome-wide study supported FP by placing Psettodes, the earliest-diverging flatfish lineage, among symmetric relatives within Carangaria, the clade that also includes billfishes, jacks, mahi-mahi, and barracudas. Subsequent work traced this to base-composition artifacts and inadequate substitution modeling. Here we revisit the question using whole-genome phylogenomic and synteny data from 17 carangarian species spanning flatfishes and carangarian relatives. We contribute three new chromosome-level assemblies, including the first for Psettodes. Nucleotide-based coalescent analyses (e.g., ROADIES, CASTER) yield strong support for FM, with Psettodes sister to all other flatfishes. Microsynteny analyses built from conserved gene-order blocks corroborate this result: topology tests, cluster-profile counts, and rearrangement-based trees favor FM over two competing FP topologies. Macrosynteny, based on chromosome-scale rearrangements, yields a more mixed signal, with support for FM depending on the metric and taxon-sampling scheme. We interpret this scale-dependent pattern in the context of the explosive post-Cretaceous radiation of Carangaria. The short intervals between speciation events that characterize rapid radiations appear to have left sufficient signal in fine-grained microsyntenic rearrangements, while chromosome-scale rearrangements were too rare to consistently resolve these closely spaced splits. When integrated with evidence from conserved developmental mechanisms active during metamorphosis, the stage at which flatfish asymmetry first emerges, and from the exceptionally complete fossil record, our multi-scale genomic evidence supports a single evolutionary origin of flatfish asymmetry.

evolutionary biology↗

bioRxiv2026

Efficient genomic prediction at reduced training size and moderate marker density in an expanded aus-NAM population of rice

Genomic selection (GS) can accelerate genetic gain in crops, but its effectiveness depends on training population design and marker density. Nested association mapping (NAM) populations provide a structured framework that captures broad allelic diversity within a controlled genetic background. Here, we evaluated genomic prediction (GP) and genome-wide association study (GWAS) performance in an expanded aus-NAM population of rice comprising 1,818 recombinant inbred lines across 14 families and 11 agronomic traits, using genotyping-by-sequencing (GBS) markers and projected whole-genome sequence variants. Prediction accuracy plateaued at moderate marker densities ([~]20k SNPs) and with training populations of [~]500 lines ([~]40-60% of the available pool), with trait heritability emerging as the strongest determinant of predictive performance rather than model choice or marker density. In contrast, GWAS resolution continued to improve with increasing marker density, enabling detection of additional loci, including a chromosome 12 locus associated with heading date, while consistently recovering well-characterized genes such as EARLY HEADING DATE 1 (Ehd1) and SEMIDWARF 1 (SD1). These contrasting patterns indicate that GP reaches near-optimal performance once genome-wide variation is adequately represented, whereas GWAS benefits from higher marker density through improved locus resolution. The present study establishes a benchmark for implementing breeding programs involving japonica/indica crosses using GP in a single environment.

plant biology↗

bioRxiv2026

Precision-Based Filtering Facilitates Integration of Conventional and Single-Nucleus Transcriptomes to Identify Time- and Temperature-Sensitive Cell Populations

Transcriptome analysis via RNA sequencing (RNAseq) has become a ubiquitous method of molecular characterization from whole organisms, dissected tissues, and single cells. These experiments have provided an extraordinary volume of data describing the molecular states and responses to many conditions. However, standard approaches to RNAseq analysis commonly use expression level filters that eliminate potentially useful data in the service of decreasing noise. Here we describe the implementation of a coefficient of variation-based filter for RNAseq gene expression data. This filter prioritizes consistent data across replicates, allowing lowly-expressed genes with low-variation measurements to be retained for downstream analysis. We show that, in our Arabidopsis RNAseq data set, this filter allows for the inclusion of many more transcription factors than even a low-stringency expression level filter. We find that these lowly-expressed genes mark specific cell clusters in our single-nucleus (sn)RNAseq dataset. We further characterize communities of co-expressed genes, sampled across the day at two growth temperatures, in relation to snRNAseq cell clusters, finding evidence for a highly photosynthetic cell population, and a cell state marked by high cell division and translation. These methods can be expanded to RNAseq analysis in many systems, facilitating the construction of more detailed models of tissue-specific gene regulatory networks.

bioinformatics↗

bioRxiv2026

The circadian clock gates lateral root development

Lateral root (LR) formation remodels root architecture, yet how temporal information integrates with hormonal cues remains unclear. We show that the circadian clock component EARLY FLOWERING 3 (ELF3) acts as a temporal gatekeeper of root organogenesis in Arabidopsis thaliana. Time-resolved transcriptomics, imaging, and genetics reveal that ELF3 restrains hormone-induced pericycle proliferation by maintaining rhythmic expression of key regulators under callus-inducing conditions. Loss of ELF3 disrupts rhythmic gene expression, enhancing callus growth and accelerating LR development. ELF3 functions through its target LNK1 to regulate the MADS-box genes AGL14 and AGL20. Notably, callus-inducing signals partially restore rhythmicity in elf3 mutants, revealing feedback from hormonal signaling to the circadian system. These findings identify ELF3 as an integrator of circadian and hormonal inputs that temporally constrains root developmental plasticity.

plant biology↗

bioRxiv2025

Pangenome of U.S. ex-PVP and Wild Sorghum Reveals Structural Variants and Selective Sweeps Shaping Adaptation and Trait Improvement

Sorghum is a cereal crop grown for food, feed, and biofuel, yet the genomic diversity of elite commercial lines remains underexplored. Here, we assembled a sorghum pangenome from long-read sequences of 46 U.S. ex-Plant Variety Protection Act (ex-PVP) cultivars and a haplotype-resolved assembly of one wild accession. The pangenome revealed largely conserved genomic architecture across elite lines but identified presence-absence variations (PAVs) in 28% of gene families, including genes involved in stress responses and starch metabolism, such as STARCH BRANCHING ENZYME I (SBE1). Selective sweep analyses detected strong signals at key flowering-time loci (Ma1/SbPRR37, Ma2, Ma6) and circadian/light regulators (PHOT1, DET1, XAP5), with PHYA showing copy number variations (CNVs). Analysis of 24 wild accessions, beyond the core pangenome panel, detected CNVs in MULTIDRUG AND TOXIC COMPOUND EXTRUSION (MATE) transporters, potentially reflecting selection or breeding for enhanced herbicide detoxification. These results highlight breeding-shaped regions and position the pangenome as a resource for sorghum improvement.

genomics↗

bioRxiv2025

Transcriptomic stress responses in Vaccinium spp. F1 hybrids: Implications for temperature-resilient cranberry breeding using a crop wild relative

O_LICranberry (Vaccinium macrocarpon) is an important North American fruit crop with vulnerability to temperature extremes and a relatively recent domestication history. Hybridization with a cold-adapted crop wild relative (CWR), V. oxycoccos, offers a strategy for improving temperature stress tolerance. C_LIO_LIWe conducted RNA sequencing (RNA-seq) on V. macrocarpon and F1 hybrids between V. macrocarpon and V. oxycoccos subjected to acute heat and cold stress, capturing early transcriptional responses up to 30 minutes (heat) and 95 minutes (cold) after treatment onset. We then evaluated differences in responses across genotypes and stress conditions. C_LIO_LIDifferential expression analysis and functional profiling revealed cold-induced differences in pathways related to photosynthesis, ribosomes, defense, and hormone signaling. No subgenome-specific functional specialization was observed. Two F1 hybrids exhibited suggestive cold resilience, with expression changes elevated at 60 minutes but declining by 95 minutes. Hybrids also displayed substantial regulatory variation under stress and transgressive downregulation of photosynthesis genes under ambient conditions. C_LIO_LIThese findings suggest that V. oxycoccos introgression could be utilized in breeding cold-tolerant cranberry cultivars. Variation observed between F1 hybrids reflects the diversity introduced through CWR germplasm and provides opportunities for selection. Conservation of V. oxycoccos and other CWRs remains critical for future crop improvement. C_LI Plain Language SummaryCranberries face increasing threats from temperature extremes. We explored crossbreeding cultivated cranberry (Vaccinium macrocarpon) and a cold-adapted wild relative (V. oxycoccos) to improve resilience to temperature stress. Hybrids showed different gene expression patterns from their parents, especially under cold stress. Our findings support the value of wild relatives in crop improvement and emphasize the importance of protecting wild plant diversity.

plant biology↗

bioRxiv2025

Domoic acid biosynthesis and genome expansion in Nitzschia navis-varingica

Production of the neurotoxin domoic acid (DA) by benthic diatom Nitzschia navis-varingica poses considerable health and economic concerns. In this study, we employed whole genome sequencing and transcriptomic analyses of regionally distinct N. navis-varingica strains to unravel the genomic underpinnings of DA biosynthesis. Our analyses revealed sizable genomes--characterized by an abundance of repetitive elements and noncoding DNA--that exceed the size of any other pennate diatoms. Central to our findings is the discovery of an expanded domoic acid biosynthesis (dab) gene cluster, spanning over 60 kb and marked by a unique organization that includes core genes interspersed with additional genetic elements. Phylogenetic and syntenic comparisons indicate that transposition events may have driven the expansion and reorganization of this cluster. Biochemical assays validated that the kainoid synthase encoded by dabC catalyzes the formation of isodomoic acid B, thereby establishing a distinct chemotype in contrast to the DA profiles of planktonic diatoms. These results highlight the evolutionary trajectory of DA biosynthesis in diatoms and potential advantages conferred by genome expansion and enzyme diversification in dynamic marine environments. IMPORTANCEDomoic acid (DA) is a potent neurotoxin produced by marine micro- and macroalgae problematic to fisheries and toxic to humans and animals. Our study elucidates the molecular mechanisms underlying DA production in the widespread Western Pacific benthic diatom, Nitzschia navis-varingica. Genomic and biochemical insights add information to our understanding of the evolution of toxin production across diverse phyla and also fill a gap in the knowledge of secondary metabolism in marine diatoms. These findings provide a genetic framework for identifying toxin production and its impacts in the benthos of vulnerable, coastal ecosystems.

genomics↗

bioRxiv2025

Triploidy is prominent in the duckweed Lemna minor complex

Duckweeds (Lemnaceae Martinov) are aquatic monocotyledonous flowering plants comprising five genera and 35 recognized species, known for being the smallest and fastest-growing flowering plants on Earth. Many species are morphologically indistinguishable due to their highly reduced structures, yet molecular evidence suggests that visually similar clones may represent distinct species or hybrids. For example, clonal accessions of the globally distributed Lemna minor in the Landolt Duckweed Collection exhibit genome size variations of several hundred megabases (Mb), raising questions about their taxonomic classification and evolutionary origins. We analyzed 58 presumed L. minor clones to resolve these relationships using a comprehensive suite of methods, including whole-genome sequencing (WGS), flow-cytometric genome size measurements, molecular markers, chromosome counting, and genomic in situ hybridization (GISH). Our findings reveal extensive genome plasticity within the "Lemna minor complex," identifying diploid and triploid L. minor clones, as well as di-haploid and triploid interspecific hybrids called L. x japonica (L. minor x L. turionifera), L. x mediterranea (L. minor x L. gibba), and a novel African-clade distinct from known L. minor lineages. Triploidy was prevalent, occurring in 29% of the clones, and was associated with enhanced growth under optimal conditions but reduced performance under high light and temperature. These findings highlight the widespread role of triploidy, cryptic species, and hybridization in the L. minor complex, emphasizing the importance of multiple approaches for accurately classifying duckweed species and understanding their evolutionary trajectories.

plant biology↗

bioRxiv2025

Enabling the Study of Gene Function in Gymnosperms: VIGS in Ephedra tweedieana

PremiseAs the sister clade to angiosperms, gymnosperms are key to enabling the reconstruction of ancestral gene regulatory networks for seed plants. However, tools to rapidly and efficiently investigate gene function in gymnosperms remain limited due to the challenges of long life cycles and large genome sizes. Species within the xerophytic genus Ephedra (Gnetales) have comparatively smaller genomes and shrubby growth habits with shorter life spans, making them better suited for greenhouse cultivation and laboratory experiments. Methods and ResultsHere, we implement Virus-Induced Gene Silencing (VIGS) to manipulate gene expression in Ephedra tweedieana. Agrobacterium-mediated vacuum infiltration of Tobacco Rattle Virus (TRV2 and TRV1) in seedlings resulted in highly efficient silencing of the E. tweedieana PHYTOENE DESATURASE ortholog EtwPDS. The expected photobleaching phenotype was observed as early as two weeks. It lasted at least three months, in stems, shoot tips, leaves, axillary meristems, and lateral branches of treated plants. ConclusionsThis first report of transient transformation and targeted gene silencing in a gymnosperm will further enable functional studies of the genetic mechanisms underpinning adaptations in this important and underrepresented lineage of seed plants.

plant biology↗

bioRxiv2025

Complete genome sequence assembly elucidates evolution and regulation of 5S rDNA loci in the greater duckweed Spirodela polyrhiza

We resolved the molecular architecture of the 5S rDNA loci in an aquatic monocot Spirodela polyrhiza. Measurements of fluorescence in situ hybridization signals revealed two loci with 5S rDNA clusters. A combination of the extra-long DNA reads and conventional sequencing of 5S rDNA repeats allowed the assembling of complete loci sequences located on ChrSp6 and ChrSp13. The homologous chromosomes of the ChrSp6 locus contain clusters of 40 and >60 copies of 5S rDNA repeat units with intergenic spacer (IGS) of 400 bp. The ChrSp13 locus is represented by 5S rDNA clusters of rRNA genes spaced by IGSs of 1,056 or 1,069 bp arranged in two sub-clusters, suggesting a different rate of repeat homogenization between loci. The G/C-rich 5S rDNA arrays in both loci are embedded in A/T-enriched chromosome regions with possible regulatory functions. The TATA-like boxes of the 5S rDNA repeat on ChrSp6 and ChrSp13 exhibit different affinities for the TATA-binding protein in 3D modeling of protein/DNA interactions, suggesting a locus-specific regulation of rRNA transcription. Our findings shed light on the molecular architecture of the 5S rDNA loci, which could foster an innovative era for the principles of evolution and regulation of rRNA genes in plants.

molecular biology↗

bioRxiv2024

The evolution of heteromorphic sex chromosomes in plants

Sex chromosomes in cannabis and hop were identified a century ago because of their obvious visible differences in size (heteromorphy). However, we know little about the genes they contain that control the development of the inflorescences. Here we assembled genomes, with phased sex chromosomes, for hop and cannabis. The XY chromosomes share an origin prior to the divergence between the genera >36 MYA. Due to the inheritance patterns of the XYs, the male-specific region of the Y is highly-degenerated, with substantial gene loss, while the X shows faster rates of molecular evolution. Consistent with the theory that these species lack an active-Y system, no clear sex-determining genes reside on the Y. Instead, an X-linked homolog of aminocyclopropane-1-carboxylate synthase (ACS), that is involved in the ethylene biosynthesis pathway, determines the fate of the female inflorescence. Beyond sex determination, the sex chromosomes contribute to the sexual dimorphism in ecology and physiology and have played a role in the domestication and breeding of these species.

evolutionary biology↗

bioRxiv2024

FT-like genes in Cannabis and hops: sex specific expression and copy-number variation may explain flowering time variation

Cannabis sativa is a fascinating, yet under-researched, species. To facilitate the global expansion of C. sativa cultivation a greater understanding of flowering time control is crucial. The PEBP gene family consists of universal promoters and repressors of flowering, with homologs of FLOWERING LOCUS T (FT) being highly conserved key regulators of flowering. FT encodes florigen, and balancing the florigen and anti-florigen signals is key for fine-tuning a crops flowering to local climatic conditions. Here, we provide an in-depth characterisation of the PEBP gene family in C. sativa and the closely related species H. lupulus. Phylogenetic analysis reveals expansion of FT and TFL1/CEN clades in the Cannabaceae. The retention of the duplicated PEBP genes likely has functional significance with divergent sequences and expression patterns hinting at signatures of sub-functionalisation. We speculate that duplicated PEBP genes have been crucial to the evolution of photoperiod insensitivity and sexually dimorphic flowering in C. sativa and harnessing the available genetic variation for these traits will be key for establishing this future crop.

plant biology↗

bioRxiv2024

Soil depth determines the microbial communities in Sorghum bicolor fields

Sorghum bicolor, an important global crop, adapted to thrive in hotter and drier conditions than maize or rice, has deep roots that interact with a unique and stratified soil microbiome that plays a crucial role in plant health, growth, and carbon storage. Microbiome studies on agricultural soils, particularly fields growing S. bicolor, have been mostly limited to surface soils (<30 cm). Here we investigated the abiotic factors of soil properties, field location, depth, and the biotic factors of sorghum type across 38 genotypes on the soil microbiome. Utilizing 16S rRNA gene amplicon sequencing, our analysis reveals significant changes in microbial composition and decreasing diversity at increasing soil depths within S. bicolor regardless of genotype or fields. Notably, specific microbial families, such as Thermogemmatisporaceae and an unclassified family within the ABS-6 order, were enriched in deeper soil layers beyond 30 cm. Additionally, microbial richness and diversity declined with depth, reaching a minimum at the 60 - 90 cm layer, with layers beyond 90 cm increasing in alpha diversity. These findings highlight the importance of soil depth in agricultural soil microbiome studies.

microbiology↗

bioRxiv2024

OrthoBrowser: Gene Family Analysis and Visualization

The analysis of gene families across diverse species is pivotal in elucidating evolutionary dynamics and functional genomic landscapes. Typical analysis approaches often require significant computational expertise and user time. We introduce OrthoBrowser, a static site generator that will index and serve phylogeny, gene trees, multiple sequence alignments, and novel multiple synteny alignments. This greatly enhances the usability of tools like OrthoFinder by making the detailed results much more visually accessible. This interface can scale reasonably up to hundreds of genomes, allows a user to filter this large dataset to a subset of samples they are interested in at that particular moment in time, or "zoom in" to only a subtree of the orthogroup. The multiple synteny alignment method uses a progressive hierarchical alignment approach in the protein space using orthogroup membership to establish orthology. Orthobrowser makes it easy for users to identify, interact with, explore, and share key information about their gene families of interest. AvailabilityOrthoBrowser is available under the MIT license at: https://gitlab.com/salk-tm/orthobrowser Contactnhartwick@salk.edu Supplementary InformationExample OrthoBrowser Results: https://orthobrowserexamples.netlify.app/

bioinformatics↗

bioRxiv2024

Domesticated cannabinoid synthases amid a wild mosaic cannabis pangenome

Cannabis sativa is a globally significant seed-oil, fiber, and drug-producing plant species. However, a century of prohibition has severely restricted legal breeding and germplasm resource development, leaving potential hemp-based nutritional and fiber applications unrealized. Existing cultivars are highly heterozygous and lack competitiveness in the overall fiber and grain markets, relegating hemp to less than 200,000 hectares globally1. The relaxation of drug laws in recent decades has generated widespread interest in expanding and reincorporating cannabis into agricultural systems, but progress has been impeded by the limited understanding of genomics and breeding potential. No studies to date have examined the genomic diversity and evolution of cannabis populations using haplotype-resolved, chromosome-scale assemblies from publicly available germplasm. Here we present a cannabis pangenome, constructed with 181 new and 12 previously released genomes from a total of 156 biological samples from both male (XY) and female (XX) plants, including 42 trio phased and 36 haplotype-resolved, chromosome-scale assemblies. We discovered widespread regions of the cannabis pangenome that are surprisingly diverse for a single species, with high levels of genetic and structural variation, and propose a novel population structure and hybridization history. Conversely, the cannabinoid synthase genes contain very low levels of diversity, despite being embedded within a variable region containing multiple pseudogenized paralogs and distinct transposable element arrangements. Additionally, we identified variants of acyl-lipid thioesterase (ALT) genes2 that are associated with fatty acid chain length variation and the production of the rare cannabinoids, tetrahydrocannabinol varin (THCV) and cannabidiol varin (CBDV). We conclude the Cannabis sativa gene pool has only been partially characterized, and that the existence of wild relatives in Asia remains likely, while its potential as a crop species remains largely unrealized.

plant biology↗