G3: Genes|Genomes|Genetics最新文献

Insects rely on the translation of environmental chemical cues into behaviors necessary for survival and reproduction. Specific chemosensory receptors belonging to the odorant and gustatory receptor groups detect odorant and gustatory cues, respectively, making them crucial to these processes. How odorant (OR) and gustatory (GR) receptor expression profiles change in combination with changing life strategies is not well understood. Using genomic and transcriptomic resources we annotated the OR and GR expression profiles across all life stages of the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, a major pest of corn in the US and Europe. Genomic analyses identified 193 ORs and 189 GRs, of which 125 and 116 were found to be expressed, respectively, in one or more WCR life stages. WCR larvae are subterranean and feed on roots before emerging as adults aboveground. Expression profile analyses revealed first instar larvae possess a unique OR and GR repertoire distinct from other instars and adults, suggesting a role in host plant finding. Similarly, a subset of ORs and GRs differed in their expression levels between adult male and female antennae. By comparing the phylogenetic relationship of ORs and GRs, we identified several receptors with potentially important roles in WCR foraging and reproductive behavior. Together, this study provides support for future investigations into the ecology and evolution of chemoreception in insects.

The rapid advancement of genomic technologies has enabled the production of highly contiguous reference genomes for non-model organisms. However, these methods often require exceptionally fresh material containing unfragmented high molecular weight nucleic acids. Researchers who preserve field-collected specimens in ethanol at ambient temperatures, prior to transferring them to long-term frozen archives, face challenges in applying advanced genomic approaches due to DNA and RNA fragmentation under suboptimal preservation conditions. To explore the potential of such preserved specimens as sources of reference genomes, we utilized Nanopore MinION technology to generate genomic data from a frozen archived specimen of the endemic alpine ground beetle Carabus (Platycarabus) depressus. Using a rapid in-house protocol for high molecular weight DNA extraction, followed by sequencing on a single flow cell, we produced 8.75 million raw reads with an N50 of 2.8 Kb. The resulting assembly achieved remarkable completeness, recovering up to 98% of BUSCO genes, despite a moderate N50 of 945 Kb. This genome is only the second available for the taxonomically diverse genus Carabus and highlights the feasibility of performing short-to-long-read sequencing from frozen archived specimens commonly housed in natural history collections. These findings open new avenues for advancing non-model organism genomics and its downstream applications.

Collective cell migration is critical to embryonic development, wound healing, and the immune response, but also drives tumor dissemination. Understanding how cell collectives coordinate migration in vivo has been a challenge, with potential therapeutic benefits that range from addressing developmental defects to designing targeted cancer treatments. The small GTPase Rap1 has emerged as a regulator of both embryogenesis and cancer cell migration. How active Rap1 coordinates downstream signaling functions required for coordinated collective migration is poorly understood. Drosophila border cells undergo a stereotyped and genetically tractable in vivo migration within the developing egg chamber of the ovary. This group of 6-8 cells migrates through a densely packed tissue microenvironment and serves as an excellent model for collective cell migration during development and disease. Rap1, like all small GTPases, has distinct activity state switches that link extracellular signals to organized cell behaviors. Proper regulation of Rap1 activity is essential for successful border cell migration yet the signaling partners and other downstream effectors are poorly characterized. Using the known requirement for Rap1 in border cell migration, we conducted a dominant suppressor screen for genes whose heterozygous loss modifies the migration defects observed upon constitutively active Rap1V12 expression. Here we identified seven genomic regions on the X chromosome that interact with Rap1V12. We mapped three genomic regions to single Rap1-interacting genes, frizzled 4, Ubiquitin specific protease 16/45, and strawberry notch. Thus, this unbiased screening approach identified multiple new candidate regulators of Rap1 activity with roles in collective border cell migration.

Work in many systems has shown large-scale changes in gene expression during aging. However, many studies employ just two, arbitrarily-chosen timepoints at which to measure expression, and can only observe an increase or a decrease in expression between "young" and "old" animals, failing to capture any dynamic, non-linear changes that occur throughout the aging process. We used RNA sequencing to measure expression in male head tissue at 15 timepoints through the lifespan of an inbred Drosophila melanogaster strain. We detected >6,000 significant, age-related genes, nearly all of which have been seen in previous Drosophila aging expression studies, and which include several known to harbor lifespan-altering mutations. We grouped our gene set into 28 clusters via their temporal expression change, observing a diversity of trajectories; some clusters show a linear change over time, while others show more complex, non-linear patterns. Notably, re-analysis of our dataset comparing the earliest and latest timepoints - mimicking a two-timepoint design - revealed fewer differentially-expressed genes (around 4,500). Additionally, those genes exhibiting complex expression trajectories in our multi-timepoint analysis were most impacted in this re-analysis; their identification, and the inferred change in gene expression with age, was often dependent on the timepoints chosen. Informed by our trajectory-based clusters, we executed a series of gene enrichment analyses, identifying enriched functions/pathways in all clusters, including the commonly seen increase in stress- and immune-related gene expression with age. Finally, we developed a pair of accessible Shiny apps to enable exploration of our differential expression and gene enrichment results.

Identifying the genetic architecture of complex traits requires access to populations with sufficient genetic diversity and recombination. Multiparent Advanced Generation InterCross (MAGIC) populations are a powerful resource due to their balanced population structure, allelic diversity, and enhanced recombination. However, implementing a MAGIC population in complex polyploids such as wheat is challenging, as wheat harbors many introgressions, inversions, and other genetic factors that interfere with linkage mapping. By utilizing a comprehensive crossing strategy, additional rounds of mixing, and novel genotype calling approaches, we developed a bread wheat 8-parent MAGIC population of over 3000 genotyped recombinant inbred lines derived from 2151 distinct crosses. This effort resulted in a dense genetic map covering the complete genome. Further rounds of inter-crossing led to increased recombination in inbred lines, as expected. We identified structural variation highlighted by segregation distortion, along with epistatic allelic interactions between specific founders. We report on a novel and effective resource for genomic and trait exploration in hexaploid wheat, capable of detecting small genetic effects and epistatic interactions due to the high level of recombination and large number of lines. The interactions and genetic effects identified provide a basis for ongoing research to understand the basis of allelic frequencies across the genome, particularly where economically important loci are involved.

Numerous studies have revealed a signature of strong adaptive evolution in the piwi-interacting RNA (piRNA) machinery of Drosophila melanogaster, but the cause of this pattern is not understood. Several hypotheses have been proposed. One hypothesis is that transposable element (TE) families and the piRNA machinery are co-evolving under an evolutionary arms race, perhaps due to antagonism by TEs against the piRNA machinery. A related, though not co-evolutionary, hypothesis is that recurrent TE invasion drives the piRNA machinery to adapt to novel TE strategies. A third hypothesis is that ongoing fluctuation in TE abundance leads to adaptation in the piRNA machinery that must constantly adjust between sensitivity for detecting new elements and specificity to avoid the cost of off-target gene silencing. Rapid evolution of the piRNA machinery may also be driven independently of TEs, and instead from other functions such as the role of piRNAs in suppressing sex-chromosome meiotic drive. We sought to evaluate the impact of TE abundance on adaptive evolution of the piRNA machinery in D. melanogaster and 2 species with higher repeat content-Drosophila ananassae and Drosophila willistoni. This comparison was achieved by employing a likelihood-based hypothesis testing framework based on the McDonald-Kreitman test. We show that we can reject a faster rate of adaptive evolution in the piRNA machinery of these 2 species. We propose that the high rate of adaptation in D. melanogaster is either driven by a recent influx of TEs that have occurred during range expansion or selection on other functions of the piRNA machinery.

The RNA exosome is an evolutionarily conserved, multiprotein complex that is the major RNase in 3' processing and degradation of a wide range of RNAs in eukaryotes. Single amino acid changes in RNA exosome subunits cause rare genetic diseases collectively called exosomopathies. However, distinguishing disease-causing variants from non-pathogenic ones remains challenging, and the mechanism by which these variants cause disease is largely unknown. Previous studies have employed a budding yeast model of RNA exosome-linked diseases that relies on mutating the orthologous yeast genes. Here, we develop a humanized yeast model of exosomopathies that allows us to unambiguously assess damaging effects of the exact patient variant in budding yeast. Individual replacement of the yeast subunits with corresponding mammalian orthologs identified six out of nine non-catalytic core subunits of the budding yeast RNA exosome that can be replaced by a mammalian subunit, with three of the replacements supporting close to normal growth. Further analysis of the disease-associated variants utilizing the hybrid yeast/mammalian RNA exosome revealed functional defects caused by both previously characterized and uncharacterized variants of EXOSC2, EXOSC4, EXOSC7, and EXOSC9. Analysis of the protein levels of these variants indicates that a subset of the patient-derived variants causes reduced protein levels, while other variants are defective but are expressed as well as the reference allele, suggesting a more direct contribution of these residues to RNA exosome function. This humanized yeast model of exosomopathies provides a convenient and sensitive genetic tool to help distinguish damaging RNA exosome variants from benign variants. This disease model can be further exploited to uncover the underpinning mechanism of RNA exosome defects.

The Ga2 locus controls unilateral cross incompatibility in Zea species. Molecular characterization of this locus indicates that fully functional loci contain two components that function in male and female reproductive tissues, Ga2P and Ga2F respectively. Some Zea mays varieties have been reported to be capable of overcoming the Ga2 reproductive barrier including the widely used inbred line Mo17. Our objective was to better understand the structure, function and distribution of the Ga2 locus in Zea species. To do this, we first examined the ability to overcome the Ga2 reproductive barrier across a diverse sample of Zea mays inbred lines. The ability of Mo17 to overcome the Ga2 reproductive barrier was mapped to the Ga2 locus. We next examined the structure of the Ga2 locus. Like the Ga1 locus, the Ga2 locus in Zea mays sp. contains multiple repeated functional genes and pseudogenes encoding the Ga2P and Ga2F pectin methylesterase enzymes. The organization of these repeat units is similar in the three sweet corn lines examined, and all are capable of overcoming the reproductive barrier. The non-sweet corn lines have a variety of genome structures, but these structures do not correlate well with the ability to overcome the Ga2 reproductive barrier. Levels of transcripts derived from Ga2P and the ability to overcome the reproductive barrier in sweet corn are higher in sweet corn lines than the other lines tested. The role of Ga2P in Zea mays in the apparent absence of a functional female factor remains unclear and may be evidence for an additional role in reproductive tissues.

The European green toad (Bufotes viridis) is geographically widely distributed. While the species global conservation status is labeled as of least concern by the IUCN, it is declining in many parts of its range where populations are fragmented and isolated. A high-quality reference genome is an important resource for conservation genomic researchers who are trying to understand and interpret the genomic signals of population decline, inbreeding, and the accumulation of deleterious mutations. Here, we assembled and annotated a chromosome-level reference genome for B. viridis as part of the European Reference Genome Atlas pilot project. The genome assembly, with a size of ∼3.89 Gb consists of 11 chromosomes and an additional 2,096 unplaced scaffolds. The final assembly had a scaffold N50 value of 478.39 Mb and covered 90.4% single copy tetrapod orthologs, and 46.7% repetitive elements. Finally, a total of 23,830 protein-coding genes matching a known gene, together with 56,974 mRNAs were predicted. This high-quality reference genome will benefit amphibian evolutionary genomics research and enable conservation genetic studies to inform practical conservation work on this species.

Facultatively symbiotic corals provide important experimental models to explore the establishment, maintenance, and breakdown of the mutualism between corals and members of the algal family Symbiodiniaceae. Here, we report the de novo chromosome-scale genome assembly and annotation of the facultatively symbiotic, temperate coral Astrangia poculata. Though widespread segmental/tandem duplications of genomic regions were detected, we did not find strong evidence of a whole genome duplication (WGD) event. Comparison of the gene arrangement between A. poculata and the tropical coral Acropora millepora revealed considerable conserved colinearity despite ∼415 million years of divergence. Gene families related to sperm hyperactivation and innate immunity, including lectins, were found to contain more genes in A. millepora relative to A. poculata. Sperm hyperactivation in A. millepora is expected given the extreme requirements of gamete competition during mass spawning events in tropical corals, while lectins are important in the establishment of coral-algal symbiosis. By contrast, gene families involved in sleep promotion, feeding suppression, and circadian sleep/wake cycle processes were expanded in A. poculata. These expanded gene families may play a role in A. poculata's ability to enter a dormancy-like state ("winter quiescence") to survive freezing temperatures at the northern edges of the species' range.