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Open Access Publications from the University of California

Scholarly Works (10 results)

  • Thesis
  • Peer Reviewed
UC Berkeley Electronic Theses and Dissertations (2015)

Duplication has long been recognized as an evolutionary source of novelty. The relaxation of purifying selection following duplication allows for normally deleterious mutations to persist long enough to give rise to novel phenotypes. Whole-genome duplications (WGDs) are a specific type of duplication, in which a species suddenly finds itself with two copies of all of its genomic loci. While the fate of most of the duplicated loci is to be lost, those that persist are thought to underlie the innovations seen in groups with a history of polyploidy, such as flowering plants, yeast, Paramecium, and vertebrates. These ancient events give us an idea of how WGDs can drive the radiation of large and diverse phyla, but do not give us any information on the genomic response immediately following polyploidy. This thesis provides insights into the origins of polyploidy and its effects on genome dynamics.

There are two models for the mechanism of polyploidy: autopolyploidy and allopolyploidy. Autopolyploids are formed by doubling the somatic chromosomes in the zygote or early embryo. Allopolyploids are formed by the hybridization of two related, but genetically distinct, species, followed by chromosome doubling. If there are no extant diploid relatives, it can be difficult to distinguish between these two models. One feature of allopolyploids is the lack of recombination between their homeologous chromosomes. The end result is that any markers that were unique to each species while apart, such as transposable element subfamilies, will be asymmetrically distributed on the progenitor chromosomes in an organism that recently underwent a WGD.

Xenopus laevis is an important vertebrate model in developmental and cell biology that has experienced a recent WGD (~40 million years ago [MYA], based on cDNA alignments (Hellsten, 2007). Its diploid cousin Xenopus tropicalis has become a popular genetic model frog. Comparative analysis of these two frog genomes gives us an excellent opportunity to study genome dynamics following whole genome duplication. The discovery of asymmetrically distributed transposon subfamilies supports the model that cross-species hybridization through allotetraploidy is the mechanism underlying the polyploid Xenopus radiation. Thus, the sub-genome sequence divergence of 40 MYA dates the divergence of the progenitor species, not the hybridization event. The asymmetric distribution of these elements between homeologous sequences allows us to assign chromosomes to progenitor species, named “A” and “B”, making X. laevis a unique system to study sub-genome-specific evolution. The wealth of transcriptome and epigenetic data available for Xenopus allows me to assay how these genomic changes affect gene expression as well as gene retention. The combination of these resources with genomic data gives me the resolution needed to date the hybridization both by studying the decay of unitary pseudogenes and by comparative analysis of the transposable elements discussed above.

The sub-genome from progenitor species “A” has more assembled length, longer chromosomes, a higher rate of gene retention, and higher average expression in the adult frog. The B sub-genome has higher synonymous and nonsynonymous mutation rates. The chromosomes orthologous to X. tropicalis 9 and 10 are fused in both sub-genomes of X. laevis, forming homeologous chromosomes 15 and 18, and deviate from the A/B trends discussed above. The regions of these X. laevis chromosomes orthologous to X. tropicalis chromosome 10 have a lower density of diagnostic repeats, no sub-genome bias in gene retention, and have a higher silent substitution rate. This divergence from the rest of the genome is not shared by the regions orthologous to X. tropicalis 9. I hypothesize that the short length of X. tropicalis 10 plays a role in these deviations due to a higher rate of gene conversion on shorter chromosomes.

    Cover page: Genomic Analysis of the Allotetraploid Frog, Xenopus laevis
    • Article
    • Peer Reviewed
    UC Berkeley Previously Published Works (2019)
    Size is a fundamental feature of biological systems that affects physiology at all levels. For example, the dynamic, microtubule-based spindle that mediates chromosome segregation scales to a wide range of cell sizes across different organisms and cell types. Xenopus frog species possess a variety of egg and meiotic spindle sizes, and differences in activities or levels of microtubule-associated proteins in the egg cytoplasm between Xenopus laevis and Xenopus tropicalis have been shown to account for spindle scaling [1]. Increased activity of the microtubule severing protein katanin scales the X. tropicalis spindle smaller compared to X. laevis [2], as do elevated levels of TPX2, a protein that enriches the cross-linking kinesin-5 motor Eg5 at spindle poles [3]. To examine the conservation of spindle scaling mechanisms more broadly across frog species, we have utilized the tiny, distantly related Pipid frog Hymenochirus boettgeri. We find that egg extracts from H. boettgeri form meiotic spindles similar in size to X. tropicalis but that TPX2 and katanin-mediated scaling is not conserved. Instead, the microtubule depolymerizing motor protein kif2a functions to modulate spindle size. H. boettgeri kif2a possesses an activating phosphorylation site that is absent from X. laevis. Comparison of katanin and kif2a phosphorylation sites across a variety of species revealed strong evolutionary conservation, with X. laevis and X. tropicalis possessing distinct and unique alterations. Our study highlights the diversity and complexity of spindle assembly and scaling mechanisms, indicating that there is more than one way to assemble a spindle of a particular size.
      Cover page: Kif2a Scales Meiotic Spindle Size in Hymenochirus boettgeri.
      • Article
      • Peer Reviewed
      UC Berkeley Previously Published Works (2019)
      Size is a fundamental feature of biological systems that affects physiology at all levels. For example, the dynamic, microtubule-based spindle that mediates chromosome segregation scales to a wide range of cell sizes across different organisms and cell types. Xenopus frog species possess a variety of egg and meiotic spindle sizes, and differences in activities or levels of microtubule-associated proteins in the egg cytoplasm between Xenopus laevis and Xenopus tropicalis have been shown to account for spindle scaling [1]. Increased activity of the microtubule severing protein katanin scales the X. tropicalis spindle smaller compared to X. laevis [2], as do elevated levels of TPX2, a protein that enriches the cross-linking kinesin-5 motor Eg5 at spindle poles [3]. To examine the conservation of spindle scaling mechanisms more broadly across frog species, we have utilized the tiny, distantly related Pipid frog Hymenochirus boettgeri. We find that egg extracts from H. boettgeri form meiotic spindles similar in size to X. tropicalis but that TPX2 and katanin-mediated scaling is not conserved. Instead, the microtubule depolymerizing motor protein kif2a functions to modulate spindle size. H. boettgeri kif2a possesses an activating phosphorylation site that is absent from X. laevis. Comparison of katanin and kif2a phosphorylation sites across a variety of species revealed strong evolutionary conservation, with X. laevis and X. tropicalis possessing distinct and unique alterations. Our study highlights the diversity and complexity of spindle assembly and scaling mechanisms, indicating that there is more than one way to assemble a spindle of a particular size.
        Cover page: Kif2a Scales Meiotic Spindle Size in Hymenochirus boettgeriCreative Commons 'BY' version 4.0 license
        • Article
        • Peer Reviewed
        UC Berkeley Previously Published Works (2019)
        The Western clawed frog Xenopus tropicalis is a diploid model system for both frog genetics and developmental biology, complementary to the paleotetraploid X. laevis. Here we report a chromosome-scale assembly of the X. tropicalis genome, improving the previously published draft genome assembly through the use of new assembly algorithms, additional sequence data, and the addition of a dense genetic map. The improved genome enables the mapping of specific traits (e.g., the sex locus or Mendelian mutants) and the characterization of chromosome-scale synteny with other tetrapods. We also report an improved annotation of the genome that integrates deep transcriptome sequence from diverse tissues and stages. The exon-intron structures of these genes are highly conserved relative to both X. laevis and human, as are chromosomal linkages ("synteny") and local gene order. A network analysis of developmental gene expression will aid future studies.
          Cover page: A chromosome-scale genome assembly and dense genetic map for Xenopus tropicalis
          • Article
          • Peer Reviewed
          UC Berkeley Previously Published Works (2015)
          Polyploid species have long been thought to be recalcitrant to whole-genome assembly. By combining high-throughput sequencing, recent developments in parallel computing, and genetic mapping, we derive, de novo, a sequence assembly representing 9.1 Gbp of the highly repetitive 16 Gbp genome of hexaploid wheat, Triticum aestivum, and assign 7.1 Gb of this assembly to chromosomal locations. The genome representation and accuracy of our assembly is comparable or even exceeds that of a chromosome-by-chromosome shotgun assembly. Our assembly and mapping strategy uses only short read sequencing technology and is applicable to any species where it is possible to construct a mapping population.
            Cover page: A whole-genome shotgun approach for assembling and anchoring the hexaploid bread wheat genome
            • Article
            • Peer Reviewed
            UC Berkeley Previously Published Works (2020)
            Our understanding of polyploid genome evolution is constrained because we cannot know the exact founders of a particular polyploid. To differentiate between founder effects and post polyploidization evolution, we use a pan-genomic approach to study the allotetraploid Brachypodium hybridum and its diploid progenitors. Comparative analysis suggests that most B. hybridum whole gene presence/absence variation is part of the standing variation in its diploid progenitors. Analysis of nuclear single nucleotide variants, plastomes and k-mers associated with retrotransposons reveals two independent origins for B. hybridum, ~1.4 and ~0.14 million years ago. Examination of gene expression in the younger B. hybridum lineage reveals no bias in overall subgenome expression. Our results are consistent with a gradual accumulation of genomic changes after polyploidization and a lack of subgenome expression dominance. Significantly, if we did not use a pan-genomic approach, we would grossly overestimate the number of genomic changes attributable to post polyploidization evolution.
              Cover page: Gradual polyploid genome evolution revealed by pan-genomic analysis of Brachypodium hybridum and its diploid progenitors.
              • Article
              • Peer Reviewed
              UC Berkeley Previously Published Works (2020)
              Miscanthus is a perennial wild grass that is of global importance for paper production, roofing, horticultural plantings, and an emerging highly productive temperate biomass crop. We report a chromosome-scale assembly of the paleotetraploid M. sinensis genome, providing a resource for Miscanthus that links its chromosomes to the related diploid Sorghum and complex polyploid sugarcanes. The asymmetric distribution of transposons across the two homoeologous subgenomes proves Miscanthus paleo-allotetraploidy and identifies several balanced reciprocal homoeologous exchanges. Analysis of M. sinensis and M. sacchariflorus populations demonstrates extensive interspecific admixture and hybridization, and documents the origin of the highly productive triploid bioenergy crop M. × giganteus. Transcriptional profiling of leaves, stem, and rhizomes over growing seasons provides insight into rhizome development and nutrient recycling, processes critical for sustainable biomass accumulation in a perennial temperate grass. The Miscanthus genome expands the power of comparative genomics to understand traits of importance to Andropogoneae grasses.
                Cover page: Genome biology of the paleotetraploid perennial biomass crop Miscanthus
                • Article
                • Peer Reviewed
                UC Davis Previously Published Works (2021)
                Long-term climate change and periodic environmental extremes threaten food and fuel security1 and global crop productivity2-4. Although molecular and adaptive breeding strategies can buffer the effects of climatic stress and improve crop resilience5, these approaches require sufficient knowledge of the genes that underlie productivity and adaptation6-knowledge that has been limited to a small number of well-studied model systems. Here we present the assembly and annotation of the large and complex genome of the polyploid bioenergy crop switchgrass (Panicum virgatum). Analysis of biomass and survival among 732 resequenced genotypes, which were grown across 10 common gardens that span 1,800 km of latitude, jointly revealed extensive genomic evidence of climate adaptation. Climate-gene-biomass associations were abundant but varied considerably among deeply diverged gene pools. Furthermore, we found that gene flow accelerated climate adaptation during the postglacial colonization of northern habitats through introgression of alleles from a pre-adapted northern gene pool. The polyploid nature of switchgrass also enhanced adaptive potential through the fractionation of gene function, as there was an increased level of heritable genetic diversity on the nondominant subgenome. In addition to investigating patterns of climate adaptation, the genome resources and gene-trait associations developed here provide breeders with the necessary tools to increase switchgrass yield for the sustainable production of bioenergy.
                  Cover page: Genomic mechanisms of climate adaptation in polyploid bioenergy switchgrass
                  • Article
                  • Peer Reviewed
                  UC Berkeley Previously Published Works (2016)
                  To explore the origins and consequences of tetraploidy in the African clawed frog, we sequenced the Xenopus laevis genome and compared it to the related diploid X. tropicalis genome. We characterize the allotetraploid origin of X. laevis by partitioning its genome into two homoeologous subgenomes, marked by distinct families of 'fossil' transposable elements. On the basis of the activity of these elements and the age of hundreds of unitary pseudogenes, we estimate that the two diploid progenitor species diverged around 34 million years ago (Ma) and combined to form an allotetraploid around 17-18 Ma. More than 56% of all genes were retained in two homoeologous copies. Protein function, gene expression, and the amount of conserved flanking sequence all correlate with retention rates. The subgenomes have evolved asymmetrically, with one chromosome set more often preserving the ancestral state and the other experiencing more gene loss, deletion, rearrangement, and reduced gene expression.
                    Cover page: Genome evolution in the allotetraploid frog Xenopus laevis.
                    • Article
                    • Peer Reviewed
                    UC Berkeley Previously Published Works (2016)
                    To explore the origins and consequences of tetraploidy in the African clawed frog, we sequenced the Xenopus laevis genome and compared it to the related diploid X. tropicalis genome. We characterize the allotetraploid origin of X. laevis by partitioning its genome into two homoeologous subgenomes, marked by distinct families of 'fossil' transposable elements. On the basis of the activity of these elements and the age of hundreds of unitary pseudogenes, we estimate that the two diploid progenitor species diverged around 34 million years ago (Ma) and combined to form an allotetraploid around 17-18 Ma. More than 56% of all genes were retained in two homoeologous copies. Protein function, gene expression, and the amount of conserved flanking sequence all correlate with retention rates. The subgenomes have evolved asymmetrically, with one chromosome set more often preserving the ancestral state and the other experiencing more gene loss, deletion, rearrangement, and reduced gene expression.
                      Cover page: Genome evolution in the allotetraploid frog Xenopus laevis
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