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Biology subjects

Kane, N. C.

Publications and source records attributed to Kane, N. C..

14 recordsLinked to original sources

Glaciation history and geographic barriers shape population genetic structure and diversity of Lewis flax across western North America

Factors shaping genetic variation at the continental scale include geographic barriers, glacial cycles, and genetic constraints like mating system. We examine how such factors combine to influence diversification in Linum lewisii (Lewis flax), a perennial forb native to North America. We used whole genome sequencing, spatial analyses, and demographic inference to quantify population diversity across space and time and to test expectations about mating system evolution during range expansion. We found that Lewis flax consists of four genetic clusters that diverged during the Last Glacial Period, with the Rocky Mountains acting as a dispersal barrier. These clusters arent reflected in current taxonomy. Genomic data also suggest Lewis flax is predominantly outcrossing, despite its self-compatibility. Although it is thought to have colonized North America via Beringia, we found no evidence of northern refugia. Current northern populations apparently derive from a post-glacial range expansion, and their genomic signatures may indicate a shift towards increased selfing. The mixed mating system of Lewis flax thus interacts with glacial history and geographical barriers in a way that differs from obligate outcrossing or selfing species. This study furthermore provides foundational knowledge for its use in ecological restoration and ongoing neodomestication as a perennial oilseed food crop.

evolutionary biology↗

Unraveling Cross-Cellular Communication in Cannabis sativa Sex Determination: A Network Ontology Transcript Annotation (Nota) Analysis.

Cannabis sativa L. (marijuana, hemp, cannabis) is an angiosperm species currently evolving sex chromosomes. Genetic mechanisms, primarily an XY chromosome system, dictate cannabis sex expression in dioecious populations. However, sexual expression is also governed by the interplay of hormone regulatory gene networks, influenced by both genetic and environmental factors. Within the species, some populations exhibit dioecy, monoecy, or a gradient of both. Dioecious individuals produce exclusively male or female flowers, while monoecious plants bear both male and female flowers. Remarkably, through interruption of phytohormone signal transduction via abiotic stressors, genetically male or female cannabis are able to produce flowers of the opposite sex. Previous transcriptomic analysis have identified genes associated with masculinization through the application of phytohormone signal disruption using silver thiosulfate treatment. We analyzed transcriptomic data from cannabis treated with colloidal silver to similarly induce masculinization. Using Nota (Network Ontology Transcript Annotation), a multilayer network analysis (random walk with restart) tool, we identified candidate genes involved in sex-determination. Nota and a companion program Jack facilitate multi-layer network analyses, enabling discovery and annotation of gene-trait associations. Our findings highlight Notas robust application to enrich the genetic architecture of complex traits, particularly in non-model systems like cannabis, and complex traits such as sex determination. Our analyses identified genes associated with cell wall morphogenesis and embryogenic tissue homeostasis, indicating that silver ion treatment perturbs phytohormone signal transduction through metal ion imbalance. In this reproductive strategy, cannabis is able to make use of its widely investigated sex-determining genetic architecture to navigate transient changes in co-expression and cross-cellular signaling driving embryogenic cell wall re-patterning.

systems biology↗

Understanding genetic architecture overcomes tradeoffs between seed quality and insect resistance

The sunflower (Helianthus annuus) pericarp protects the seed within from both abiotic and biotic stresses. Achenes with stronger pericarps are less susceptible to damage from insect feeding. Complicating the genetic improvement of pericarp strength is the negative correlation between pericarp thickness (a component of strength) and oil content. As breeding efforts have increased oil content, there has been a concomitant decrease in pericarp thickness. A logical sunflower improvement goal is to improve oil content while preserving pericarp strength through genetic mechanisms independent of the tradeoffs with pericarp thickness. To determine the genetic basis of oil content, pericarp strength, and thickness, we identified QTL in two populations; the Sunflower Association Mapping panel (Mandel et al., 2011) and a recombinant inbred line (RIL) population derived from a thin pericarp oilseed inbred (HA 467) crossed to a thick pericarp open pollinated variety from Turkiye (PI 170415). A region on chromosome 15 was associated with neighboring QTL for banded moth resistance, oil content, and pericarp thickness, partially underlying the trade-offs among these traits. Additional QTL on chromosome 5 and 14 for pericarp strength provide fewer trade-offs with oil content. QTL for pericarp strength on chromosome 5 and pericarp thickness on chromosome 16 were associated with large structural variants, with candidate gene presence/absence variation between the haplotypes on chromosome 5. Understanding the origin and nature of phenotypic tradeoffs is beneficial to plant biologists and sunflower breeders as they seek to understand the origin and genetic architecture of adaptive and maladaptive traits.

genetics↗

Variant filters using segregation information improve mapping of nectar-production genes in sunflower (Helianthus annuus L.)

Accurate variant calling is critical for identifying the genetic basis of complex traits, yet filters used in variant detection and validation may inadvertently exclude valuable genetic information. In this study, we compare common sequencing depth filters, used to eliminate error-prone variants associated with repetitive regions and technical issues, with a biologically relevant filtering approach that targets expected population-level Mendelian segregation. The resulting variant sets were evaluated in the context of nectar volume QTL mapping in sunflower (Helianthus annuus L.). Our previous research failed to detect a significant interval containing a strong candidate gene for nectar production (HaCWINV2). We removed certain hard filters and implemented a Chi-square goodness-of-fit test to retain variants that segregate according to expected genetic ratios. We hypothesized that this will enhance mapping resolution and capture key genetic regions previously missed. We demonstrate that biologically relevant filtering retains more significant QTL and candidate genes, including HaCWINV2, while removing variants due to technical errors more effectively, and accounted for 48.55% of phenotypic variation. In finding nine putative homologs of Arabidopsis genes with nectary function within QTL regions, we demonstrate that this filtering strategy, which considers biological contexts, has a higher power of true variant detection than the commonly used variant depth filtering strategy. PLAIN LANGUAGE SUMMARYIn genomic research, identifying genetic markers is key to understanding complex traits, but traditional methods for filtering genetic data can sometimes miss important information. In this study, we explored a new data filtering approach for mapping genes related to nectar production in sunflower. We applied a more flexible filtering method that considers how markers are expected to segregate in breeding populations. Our previous work failed to identify an important gene previously hypothesized to be involved in nectar production, likely due to overly strict filtering. Our improved approach identified nine sunflower genes related to nectar production genes in the model species Arabidopsis thaliana, as compared to zero genes identified from the previous filtering strategy. This study highlights the value of using flexible, biologically relevant filtering methods, which can lead to better results in plant genomic studies. CORE IDEASO_LIDiscovering biologically meaningful variants from sequence data requires a careful and critical view of bioinformatic workflows. C_LIO_LIThe use of arbitrary filters can remove significant genomic variation that contributes to the phenotype of interest. C_LIO_LIArbitrary filters can also fail to remove variant call errors. C_LIO_LIA Chi-square filtering strategy based on segregation ratio retained a larger number of valid variants. C_LIO_LIMore candidate regions with putative nectar-related genes and better statistical support were discovered. C_LI

plant biology↗

Environment, plant genetics, and their interaction shape important aspects of sunflower rhizosphere microbial communities

Associations with soil microorganisms are crucial for plants overall health and functioning. While much work has been done to understand drivers of rhizosphere microbiome structure and function, the relative importance of geography, climate, soil properties, and plant genetics remains unclear, as results have been mixed and comprehensive studies across many sites and genotypes are limited. Rhizosphere microbiomes are crucial for crop resistance to pathogens, stress tolerance, nutrient availability, and ultimately yield. Here we quantify the relative roles of plant genotype, environment, and their interaction in shaping soil rhizosphere communities, using 16S and ITS gene sequencing of rhizosphere soils from 10 common sunflower (Helianthus annuus) genotypes from 15 sites across the Great Plains of the United States. While site generally outweighed genotype overall in terms of effects on archaeal, bacterial and fungal richness, community composition, and taxa relative abundances, there was also a significant interaction such that genotype exerted a significant influence on archaeal, bacterial and fungal microbiomes in certain sites. Site effects were attributed to a combination of spatial distance and differences in climate and soil properties. Microbial taxa that were previously associated with resistance to the fungal necrotrophic pathogen Sclerotinia were present in most sites but differed significantly in relative abundance across sites. Our results have implications for plant breeding and agronomic microbiome manipulations for agricultural improvement across different geographic regions. ImportanceDespite the importance of plant breeding in agriculture, we still have a limited understanding of how plant genetic variation shapes soil microbiome composition across broad geographic regions. Using 15 sites across the Great Plains of North America, we show that cultivated sunflower rhizosphere archaeal, bacterial and fungal communities are driven primarily by site soil and climatic differences, but that genotype can interact with site to influence composition, especially in warmer and drier sites with lower overall microbial richness. We also show that all taxa that were previously found to be associated with resistance to the fungal pathogen Sclerotinia sclerotiorum were widespread but significantly affected by site, while a subset were also significantly affected by genotype. Our results contribute to a broader understanding of rhizosphere archaeal, bacterial and fungal community assembly, and provide foundational knowledge for plant breeding efforts and potential future microbiome manipulations in agriculture.

ecology↗

Chromosome-scale Genome Assembly of Lewis Flax (Linum lewisii Pursh.)

The shift from self-incompatibility to self-compatibility is a frequent evolutionary transition in flowering plants with numerous ecological and evolutionary consequences. It is also an advantageous transition for domestication of new crop plants, as self-compatibility makes it considerably easier to drive adaptively important alleles to fixation. In the flax genus, Linum, self-incompatibility is linked to the floral polymorphism known as heterostyly, where plants exhibit distinct floral morphs with different positioning of male and female reproductive organs. Heterostyly has been lost multiple times independently across the flax genus, leading to homostyly and self-compatibility, but the genetic causes of this transition are not fully understood. Here, we present a near telomere-to-telomere genome assembly of "Maple Grove" Lewis flax (Linum lewisii, 2n = 2x = 18), a homostylous wild blue flax species native to North America. By comparison to the genome of close heterostylous relative L. perenne, we found that the coding sequence of heterostyly candidate gene TSS1 is deleted in the Lewis flax genome, which could underlie its transition from heterostyly to homostyly. Analysis of chromosomal synteny between Lewis flax and common flax (L. usitatissimum) further revealed a striking amount of chromosomal rearrangements, which will complicate the use of comparative genomics to accelerate domestication of Lewis flax as a new perennial oilseed crop. The final, primary haplotype was 845 Mb in length and comprised 9 pseudochromosomes and 324 unplaced scaffolds with a contig N50 of 17.9 Mb. Annotation of the assembly revealed 19,593 protein-coding genes. This genomic resource will inform ongoing breeding efforts and will support its use in native ecosystem restoration and in other native plantings across western North America.

genomics↗

Gene expression and alternative splicing contribute to adaptive divergence of ecotypes

Regulation of gene expression is a critical link between genotype and phenotype explaining substantial heritable variation within species. However, we are only beginning to understand the ways that specific gene regulatory mechanisms contribute to adaptive divergence of populations. In plants, the post-transcriptional regulatory mechanism of alternative splicing (AS) plays an important role in both development and abiotic stress response, making it a compelling potential target of natural selection. AS allows organisms to generate multiple different transcripts/proteins from a single gene and thus may provide a source of evolutionary novelty. Here we examine whether variation in alternative splicing and gene expression levels might contribute to adaptation and incipient speciation of dune-adapted prairie sunflowers in Great Sand Dunes National Park, Colorado, USA. We conducted a common garden experiment to assess transcriptomic variation among ecotypes and analyzed differential expression, differential splicing, and gene coexpression. We show that individual genes are strongly differentiated for both transcript level and alternative isoform proportions, even when grown in a common environment, and that gene coexpression networks are disrupted between ecotypes. Furthermore, we examined how genome-wide patterns of sequence divergence correspond to divergence in transcript levels and isoform proportions and find evidence for both cis and trans-regulation. Together our results emphasize that alternative splicing has been an underappreciated mechanism providing source material for natural selection at micro-evolutionary time scales.

evolutionary biology↗

Genomic evidence that governmentally produced Cannabis sativa poorly represents genetic variation available in state markets

The National Institute on Drug Abuse (NIDA) is the sole producer of Cannabis for research purposes in the United States, including medical investigation. Previous research established that cannabinoid profiles in the NIDA varieties lacked diversity and potency relative to the Cannabis produced commercially. Additionally, microsatellite marker analyses have established that the NIDA varieties are genetically divergent form varieties produced in the private legal market. Here, we analyzed the genome of multiple Cannabis varieties from diverse lineages including two produced by NIDA, and we provide further support that NIDAs varieties differ from widely available medical, recreational, or industrial Cannabis. Furthermore, our results suggest that NIDAs varieties lack diversity in the single copy portion of the genome, the maternally inherited genomes, the cannabinoid genes, and in the repetitive content of the genome. Therefore, results based on NIDAs varieties are not generalizable regarding the effects of Cannabis after consumption. For medical research to be relevant, material that is more widely used would have to be studied. Clearly, having research to date dominated by a single, non-representative source of Cannabis has hindered scientific investigation.

genomics↗

Variation in mtDNA haplotypes suggests a complex history of reproductive strategy in Cannabis sativa

Cannabis is one example in angiosperms that appears to have a recent origin of dioecy and X/Y sex chromosomes. Several evolutionary explanations for this transition have been proposed, with the most parsimonious beginning with a mitochondrial mutation leading to cytoplasmic male sterility (CMS). Our study utilized 73 Cannabis sativa whole genome shotgun libraries to reveal eight different mtDNA haplotypes. The most common haplotype contained 60 of the 73 individuals studied and was composed of only dioecious individuals. However, other haplotypes contained a mix of dioecious and monoecious individuals, so haplotype alone does not predict dioecy. From these haplotype groupings we further examined the fully annotated mitochondrial genomes of four hemp individuals and looked for genetic variation affecting reproductive strategy (e.g., monoecious vs. dioecious strategies). Specifically, we searched for markers associated with CMS and for gene rearrangements within these mitochondrial genomes. Our results revealed highly syntenic mitochondrial genomes that contained approximately 60 identifiable sequences for protein coding genes, tRNAs and rRNAs and no obvious rearrangements or chimeric genes. We find no clear evidence that the different reproductive patterns are due to easily identifiable CMS mutations. Our results refute the simplest hypothesis that there was a single recent origin of dioecy in a monoecious ancestor. Instead, the story of the evolution of dioecy is likely much more complex. Further exploration of the nuclear and mitochondrial genomes and their interaction is required to fully understand Cannabis mating strategies and the possible existence of CMS in this species.

genomics↗

Aberrant regulation of RNA splicing in sunflower hybrids may underlie intrinsic incompatibilities

Alternative spicing is an integral part of gene expression in multicellular organisms that allows for diverse mRNA transcripts and proteins to be produced from a single gene. However, most existing analyses have focused on macro-evolution, with only limited research on splice site evolution over shorter term, micro-evolutionary time scales. Here we examine splicing evolution that has occurred during domestication and observe 45 novel splice forms with strongly transgressive isoform compositions, representing 0.24% of analyzed transcripts. We identify loci associated with variation in the levels of these splice forms, finding that many novel transcripts were regulated by multiple alleles with non-additive interactions. A subset of these interactions involved the expression of individual spliceosome components. These overdominant and epistatic interactions often resulted in alteration in the protein-coding regions of the transcripts, resulting in frameshifts and truncations. By associating the splice variation in these genes with size and growth rate measurements, we found that none of the individual splice variants affected these plant traits significantly, but the cumulative expression of all aberrant transcripts did show a significant reduction in growth rate associated with higher proportions of disrupted transcripts. This demonstrates the importance of co-evolution of the different spliceosomal components and their regulators and suggests that these genes may contribute to evolution of reproductive isolation as Bateson-Dobzhansky-Muller incompatibility loci. Author summaryIn multicellular organisms, it is common that segments of pre-mRNA molecules are physically removed, and the remaining segments are spliced back together. Through splicing alternative combinations of segments together, organisms produce various mRNA molecules, and thus multiple proteins, using the information encoded in a single gene. Here, we investigated the RNA of two sunflower genotypes, one wild and one domesticated, as well as the hybrid offspring resulting from a cross between the two genotypes. We found certain mRNA molecules that were spliced exclusively in the hybrids and were absent in the examined parental lines. These unique hybrid mRNAs were predicted to be consequential for the hybrids health, and thus represented a malfunction in the mechanisms that regulate splicing. These results improve our understanding of the genetic regulation of alternative splicing and how alternative splice forms evolve. Our findings may lead to further inquiries about how aberrant splicing promotes the formation of new species in nature.

evolutionary biology↗

Contrasting selection at multiple life stages maintains divergent adaptation between sunflower ecotypes

Conspecific populations living in adjacent, but contrasting, microenvironments represent excellent systems for studying natural selection. These systems are valuable because gene flow maintains genetic homogeneity except at loci experiencing strong, divergent selection. A history of reciprocal transplant and common garden studies in such systems, and a growing number of genomic studies, have contributed to understanding how selection operates in natural populations. While selection can vary across different fitness components and life stages, few studies have investigated how this ultimately affects allele frequencies and persistence of divergent populations. Here, we study two sunflower ecotypes in distinct, adjacent habitats by combining demographic models with genome-wide sequence data to estimate fitness components, absolute fitness, and allele frequency change at multiple life stages. This framework allows us to demonstrate that only local ecotypes experience positive population growth (lambda>1) and that the maintenance of divergent adaptation is mediated via habitat- and life stage-specific selection. We identify genetic variation, significantly driven by loci in chromosomal inversions, associated with different life history strategies in neighbouring ecotypes that optimize different fitness components and contribute to the persistence of each ecotype in its respective habitat.

evolutionary biology↗

Heritable associations with microbial communities are essential for necrotrophic pathogen resistance

Host-microbe interactions are increasingly recognized as important drivers of organismal health, growth, longevity, and community-scale ecological processes. However, less is known about how genetic variation affects hosts associated microbiomes and downstream phenotypes. We demonstrate that sunflower (Helianthus annuus) harbors substantial, heritable variation in microbial communities under field conditions. We show that microbial communities explain up to 77.5% of the heritable variation in resistance to root infection caused by the necrotrophic pathogen Sclerotinia sclerotiorum, and that plants grown in sterilized soil showed almost complete elimination of pathogen resistance. Association mapping revealed 69 genetic locations related to microbial abundance and Sclerotinia resistance. Although the genetic architecture is complex and quantitative, we have, in large part, elucidated previously unexplained genetic variation for resistance to this pathogen. This suggests new targets for plant breeding and demonstrates the potential for heritable microbial associations to play important roles in defense in natural and human-altered environments.

ecology↗

Gene copy number is associated with phytochemistry in Cannabis sativa

Gene copy number variation is known to be important in nearly every species where it has been examined. Alterations in gene copy number may provide a fast way of acquiring diversity, allowing rapid adaptation under strong selective pressures, and may also be a key component of standing genetic variation within species. Cannabis sativa plants produce a distinguishing set of secondary metabolites, the cannabinoids, many having medicinal utility. Two major cannabinoids -- THCA and CBDA -- are products of a three-step biochemical pathway. Using genomic data for 69 Cannabis cultivars from diverse lineages within the species, we found that genes encoding the synthases in this pathway vary in copy number, and that the cannabinoid paralogs may be differentially expressed. We also found that copy number partially explains variation in cannabinoid content levels among Cannabis plants.

genomics↗

Compromised External Validity: Federally Produced Cannabis Does Not Reflect Legal Markets

As the most widely used illicit drug, the basis of the fastest growing major industry in the US, and as a source of numerous under-studied psychoactive compounds, understanding the psychological and physiological effects of Cannabis is essential. The National Institute on Drug Abuse (NIDA) is designated as the sole legal producer of Cannabis for use in US research studies. We sought to compare the chemical profiles of Cannabis varieties that are available to consumers in states that have state-legalized use versus what is available to researchers interested in studying the plant and its effects. Our results demonstrate that the federally produced Cannabis has significantly less variety and lower concentrations of cannabinoids. Current research, which has focused on material that is far less diverse and less potent than that used by the public, limits our understanding of the plants chemical, biological, psychological, medical, and pharmacological properties. Investigation is urgently needed on the diverse forms of Cannabis used by the public in state-legal markets.

plant biology↗