Akito Y Kawahara’s research while affiliated with Florida Museum of Natural History and other places

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Publications (19)


Elevation (m) in Puerto Rico and major physiographic regions including the northern karst formations (mogotes), Sierra de Luquillo, Cajuas‐Juncos (CJ) Valley, Cordillera Central and the Sierra de Cayey.
Modelled present (1970–2000), future (2061–2080) and per cent change (%) of (a) species richness (number of species), functional richness (sum of the trait–distance matrix branch lengths), species and functional turnover (Sørensen pairwise dissimilarity index) and (b) wing length (cm), hue, saturation and brightness (colour metrics all unitless, range 0–255) for 62 butterfly species using stacked ensemble distribution models. Note the different units and scales for each panel.
Density histogram plots showing changes in modelled (a–d) species and functional richness and turnover; (e–h) wing length, hue, saturation and brightness over time. Values represent pixel‐level values calculated based on mean traits for all species with at least 65% probability of occurring in each cell. Pixels are ~1 km² spatial resolution (883 × 923 m), and (i) total area (km²) exhibiting gains or losses in each response variable.
Local regression between modelled change in area and modelled change in maximum elevation for 62 butterfly species based on current (1970–2000) and future (2061–2080) environmental conditions, showing that predicted declines in maximum elevation are not associated with changes in distribution area. In contrast, predicted increases in maximum elevation are associated with predicted increases in distribution area (R² = .35, p < .001). Histogram and density distributions for predicted change in distribution area (km²) and predicted change in maximum elevation (m) are shown along the corresponding axes. Most species with small wings were predicted to expand their habitat area and shift to higher elevations. Size and colour of data points indicate wing length (cm). Notably, species which were predicted to exhibit declines in maximum elevation were predominantly larger winged.
Temperature seasonality drives taxonomic and functional homogenization of tropical butterflies
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January 2024

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219 Reads

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3 Citations

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Aim To better understand the potential impact of climate change on butterfly assemblages across a tropical island, we model the potential for taxonomic and functional homogenization and determine climate‐ and trait‐mediated shifts in projected species distributions. Location Puerto Rico. Methods We used thousands of museum records of diurnal Lepidoptera to model current (1970–2000) and forecast future (2061–2080) species distributions and combined these to test for taxonomic and functional homogenization. We then quantified climatic‐mediated effects on current and forecasted taxonomic and functional composition and, specifically, whether temperature was a primary driver, as predicted by the temperature–size rule and the thermal melanism hypotheses. Finally, we measured wing traits important in thermoregulation (size and colour) and determined trait‐mediated changes in forecasted species distributions over time. Results Based on ensemble model outputs, taxonomic and functional richness and turnover were predicted to vary across the island's complex topography. Our models projected an increase in taxonomic and functional richness over time, and a decrease in taxonomic and functional turnover – a signature of biotic homogenization. Under future climate scenarios, models projected a decrease in wing length and an increase in wing brightness at higher elevations. One variable, temperature seasonality, was the strongest predicted driver of both the current spatial distribution and the projected per cent change over time for not only wing traits but also taxonomic and functional richness and turnover. Main conclusions The species distribution models generated here identify several priority regions and species for future research and conservation efforts. Our work also highlights the role of seasonality and climatic variability on diverse tropical Lepidoptera assemblages, suggesting that climatic variability may be an important, albeit overlooked, driver of climate change responses.

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Figure 1. Analysis of visual genes shows high conservation with day and night-flying lepidoptera with gene losses in the DAGL and innexin in M. leucophasiata.
Maximum likelihood tree of the 20 selected lepidopteran species (Representing six butterfly and eight moth families) based on 3,376 BUSCO single-copy orthologs (left), and the gene count matrix of phototransduction-related gene families (right). The diel niche of each species is indicated by orange suns (diurnal) and purple moons (nocturnal). The mean gene count (repertoire size) is shown in the colored heatmap.
Figure 2. Convergence of visual opsins in Macrosoma leucophasiata with nocturnal moth species highlighting branches under positive selection.
Opsin gene family tree showing the seven visual opsins in the studied species. Colored branches represent nocturnal (blue) and diurnal (orange) species. Stars indicate branches or nodes detected to be under positive selection by the aBSREL test for episodic diversifying selection.
New genome reveals molecular signatures of adaptation to nocturnality in moth-like butterflies (Hedylidae)

November 2023

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263 Reads

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1 Citation

Nearly all animals have a preferred period of daily activity (diel-niche), which is strongly influenced by the light environment. Sensory systems, particularly vision, are adapted to light, and evolutionary transitions to novel light environments, especially light limited ones, can impose strong constraints on eye evolution, color, and motion vision. The adaptive changes in sensory abilities of animals during these transitions, both at the genetic and neural levels, are largely unexplored. Butterflies and moths, with their diverse diel-niche shifts, are an ideal group for investigating the gene evolution linked to these transitions. While most butterflies are day-flying, hedylid butterflies are unique in being primarily nocturnal, and they represent an important evolutionary shift from diurnality to nocturnality in this clade. Here, we sequence the first high-quality Hedylidae genome and functionally annotate genes to understand genomic changes associated with shifts in diel niche. Comparing Hedylidae visual genes against day-and night-flying Lepidoptera species revealed that visual genes are highly conserved, with no major losses. However, hedylid butterfly opsins were more similar to nocturnal moths than their diurnal congeners. Tests on the evolutionary rates (dN/dS) confirmed that color vision opsins were under strong selection, similar to nocturnal moths. We propose that a convergent event of sequence evolution took place when these butterflies became nocturnal, approximately 98 million years ago.


CRISPR‐based diagnostics detects invasive insect pests

October 2023

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188 Reads

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9 Citations

Molecular Ecology Resources

Rapid identification of organisms is essential for many biological and medical disciplines, from understanding basic ecosystem processes, disease diagnosis, to the detection of invasive pests. CRISPR‐based diagnostics offers a novel and rapid alternative to other identification methods and can revolutionize our ability to detect organisms with high accuracy. Here we describe a CRISPR‐based diagnostic developed with the universal cytochrome‐oxidase 1 gene (CO1). The CO1 gene is the most sequenced gene among Animalia, and therefore our approach can be adopted to detect nearly any animal. We tested the approach on three difficult‐to‐identify moth species ( Keiferia lycopersicella , Phthorimaea absoluta and Scrobipalpa atriplicella ) that are major invasive pests globally. We designed an assay that combines recombinase polymerase amplification (RPA) with CRISPR for signal generation. Our approach has a much higher sensitivity than real‐time PCR assays and achieved 100% accuracy for identification of all three species, with a detection limit of up to 120 fM for P. absoluta and 400 fM for the other two species. Our approach does not require a sophisticated laboratory, reduces the risk of cross‐contamination, and can be completed in less than 1 h. This work serves as a proof of concept that has the potential to revolutionize animal detection and monitoring.


Phylogeny of the Poritiinae (Lepidoptera: Lycaenidae), butterflies with ant associations and unusual lichenivorous diets

January 2023

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500 Reads

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2 Citations

Systematic Entomology

The Poritiinae are a diverse subfamily of lycaenid butterflies with about 700 species divided into two major groups: the Asian endemic tribe Poritiini, and the African endemic tribe Liptenini. Among these, the Liptenini are notable for their lichenivorous diet and the strong but apparently non‐mutualistic ant associations of many species. We present the first molecular phylogeny for this subfamily, based on data from 14 gene regions, and including 218 representatives from 177 taxa (approximately 25% of species) in 50 of the 58 (86%) recognized genera. From this analysis, we confirm the division of the subfamily into two tribes, and we rearrange the Liptenini tribe into six subtribes, Durbaniina, Pentilina, Liptenina, Iridanina and Epitolina, plus a new tribe, Cooksoniina subtrib. n. , to fill a gap in the nomenclature revealed by the phylogenetic analysis. We also point to several genera in need of further taxonomic revision. Ancestral range reconstruction could not infer the range of the common ancestor of the Poritiinae; however, the common ancestor of the Poritiini was likely Asian, while that of the Liptenini was likely African, with subsequent narrowing of ranges in several lineages.



Fig. 1. (A) Subsection of a typical sticky trap collected in the field with natural bycatch using a Phthorimaea absoluta pheromone lure (no P. absoluta pictured). Each black square on the trap is approximately 25 mm 2 . (B) Subsection of a sticky trap mock community. Starting at the top going clockwise, species pictured are Anastrepha ludens, Diaphorina citri, Phthorimaea absoluta, Tribolium confusum, and Plodia interpunctella. The subsection in B is equivalent to one black square from A (approximately 25 mm 2 ).
Fig. 4. Concentration of DNA extracted from three P. interpunctella moths exposed, on sticky traps, to different temperatures over a 14-d trial period at <10% relative humidity. Moths were removed from the sticky traps with Histo-clear before DNA extraction. Raw data are available in Supp Files S2 and S3 (online only).
Results of the analysis of variance (ANOVA) showing the effect of Histo-clear concentration on PCR amplification
Real Time PCR Cq results showing little difference in re- sults between abundance, bycatch, and time of sampling
A DNA Extraction Method for Insects From Sticky Traps: Targeting a Low Abundance Pest, Phthorimaea absoluta (Lepidoptera: Gelechiidae), in Mixed Species Communities

April 2022

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276 Reads

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17 Citations

Journal of Economic Entomology

Invasive insects can cause catastrophic damage to ecosystems and cost billions of dollars each year due to management expenses and lost revenue. Rapid detection is an important step to prevent invasive insects from spreading, but improvements in detection capabilities are needed for bulk collections like those from sticky traps. Here we present a bulk DNA extraction method designed for the detection of Phthorimaea absoluta Meyrick (Lepidoptera: Gelechiidae), an invasive moth that can decimate tomato crops. We test the extraction method for insect specimens on sticky traps, subjected to different temperature and humidity conditions, and among mock insect communities left in the field for up to 21 d. We find that the extraction method yielded high success (>92%) in recovering target DNA across field and lab trials, without a decline in recovery after three weeks, across all treatments. These results may have a large impact on tomato growing regions where P. absoluta is in the early stages of invasion or not yet present. The extraction method can also be used to improve detection capabilities for other bulk insect collections, especially those using sticky traps, to the benefit of pest surveys and biodiversity studies.


Population differentiation and structural variation in the Manduca sexta genome across the United States

February 2022

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89 Reads

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7 Citations

G3 Genes Genomes Genetics

Many species that are extensively studied in the laboratory are less well characterized in their natural habitat, and laboratory strains represent only a small fraction of the variation in a species’ genome. Here we investigate genomic variation in three natural North American populations of an agricultural pest and a model insect for many scientific disciplines, the tobacco hornworm (Manduca sexta). We show that hornworms from Arizona, Kansas, and North Carolina are genetically distinct, with Arizona being particularly differentiated from the other two populations using Illumina whole-genome resequencing. Peaks of differentiation exist across the genome, but here we focus in on the most striking regions. In particular, we identify two likely segregating inversions found in the Arizona population. One inversion on the Z chromosome may enhance adaptive evolution of the sex chromosome. The larger, 8 megabase inversion on chromosome 12 contains a pseudogene which may be involved in the exploitation of a novel hostplant in Arizona, but functional genetic assays will be required to support this hypothesis. Nevertheless, our results reveal undiscovered natural variation and provide useful genomic data for both pest management and evolutionary genetics of this insect species.


Phylogeny of gracillariid leaf-mining moths: evolution of larval behaviour inferred from phylogenomic and Sanger data

October 2021

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368 Reads

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27 Citations

Cladistics

Gracillariidae is the most taxonomically diverse cosmopolitan leaf-mining moth family, consisting of nearly 2000 named species in 105 described genera, classified into eight extant subfamilies. The majority of gracillariid species are internal plant feeders as larvae, creating mines and galls in plant tissue. Despite their diversity and ecological adaptations, their phylogenetic relationships , especially among subfamilies, remain uncertain. Genomic data (83 taxa, 589 loci) were integrated with Sanger data (130 taxa, 22 loci), to reconstruct a phylogeny of Gracillariidae. Based on analyses of both datasets combined and analyzed separately , monophyly of Gracillariidae and all its subfamilies, monophyly of the clade "LAMPO" (subfamilies: Lithocolletinae, Acrocercopinae, Marmarinae, Phyllocnistinae, and Oecophyllembiinae) and relationships of its subclade "AMO" (subfamilies: Acrocercopinae, Marmarinae, and Oecophyllembiinae) were strongly supported. A sister-group relationship of Ornixolinae to the remainder of the family, and a monophyletic leaf roller lineage (Callicercops V ari + Parornichinae) + Gracillariinae, as sister to the "LAMPO" clade were supported by the most likely tree. Dating analyses indicate a mid-Cretaceous (105.3 Ma) origin of the family, followed by a rapid diversification into the nine subfamilies predating the Cretaceous-Palaeogene extinction. We hypothesize that advanced larval behaviours, such as making keeled or tentiform blotch mines, rolling leaves and galling, allowed gracillariids to better avoid larval parasitoids allowing them to further diversify. Finally, we stabilize the classification by formally re-establishing the subfamily ranks of Marmarinae stat.rev., Oecophyllembiinae stat.rev. and Parornichinae stat.rev., and erect a new subfamily, Callicercopinae Li, Ohshima and Kawahara to accommodate the enigmatic genus Callicercops.


FIG. 1.-Comparison of BUSCO (blue, yellow and red bars) and contig N50 (gray bars) results for the genomes of Trichoptera and Lepidoptera species (from top to bottom: Hydropsyche tenuis (Hydropsychidae), Stenopsyche tienmushanensis (Stenopsychidae), Hesperophylax magnus (Limnephilidae), Neomicropteryx cornuta (Micropterigidae), Tinea trinotella (Tineidae), Anthocharis cardamines (Pieridae), Autographa gamma (Noctuidae), Bombyx mori (Bombycidae)). Each silhouette to the right of the plots is derived from a member of the same genus as the genome assemblies. The phylogeny is based on Kawahara et al. (2019) and Thomas et al. (2020). Original photographs for silhouettes T. trinotella provided by Donald Hobern, Au. Gamma provide by Martin Olofsson; other silhouettes made from photographs by authors or drew by XL.
First Annotated Genome of a Mandibulate Moth, Neomicropteryx cornuta , Generated Using PacBio HiFi Sequencing

October 2021

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219 Reads

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9 Citations

Genome Biology and Evolution

We provide a new, annotated genome assembly of Neomicropteryx cornuta, a species of the so-called “mandibulate archaic moths” (Lepidoptera: Micropterigidae). These moths belong to a lineage that is thought to have split from all other Lepidoptera more than 300 million years ago and are consequently vital to understanding the early evolution of superorder Amphiesmenoptera, which contains the order Lepidoptera (butterflies and moths) and its sister order Trichoptera (caddisflies). Using PacBio HiFi sequencing reads, we assembled a highly-contiguous genome with a contig N50 of nearly 17 Mbp. The assembled genome length of 541,115,538 bp is about half the length of the largest published Amphiesmenoptera genome (Limnephilus lunatus, Trichoptera) and double the length of the smallest (Papilio polytes, Lepidoptera). We find high recovery of universal single copy orthologs with 98.1% of BUSCO genes present and provide a genome annotation of 15,643 genes aided by resolved isoforms from PacBio IsoSeq data. This high-quality genome assembly provides an important resource for studying ecological and evolutionary transitions in the early evolution of Amphiesmenoptera.


Citations (15)


... This could result in fewer organisms restricted to forest environments and their replacement by generalist or open environment species, reflecting a faunal savannisation process (Sales et al., 2020). Studies aiming to forecast the future distributions and suitable habitats of tropical insects are necessary in the face of accelerated climate and land use change (Bellaver et al., 2022;Cardoso et al., 2020;Chowdhury, 2023;Hulshof et al., 2024;Kwon et al., 2021;Salcido et al., 2020). ...

Reference:

Mixed responses among closely related Neotropical butterflies under extreme climate and land cover changes
Temperature seasonality drives taxonomic and functional homogenization of tropical butterflies

... Genome assembly and quality assessment. We assembled the genome using our previously published genome-assembly pipeline 9,10 . Briefly, raw HiFi ccs reads were fed to the Hifiasm assembler, and the assembled output was purged for duplicated haplotigs using purge_haplotigs v1.1.2 ...

New genome reveals molecular signatures of adaptation to nocturnality in moth-like butterflies (Hedylidae)

... As majority of the DNA amplification techniques require a sophisticated thermocycler, none of them are appropriate for field conditions 23,54 . CRISPR-based pest identification of insects is a promising approach under laboratory conditions and till now not been applied in the field with crude extractions; because an intricate extraction of DNA is required to successfully complete the assay 18 . As crude extract can be used as a template and LAMP reaction can be carried out on a dry bath, it is suitable for field and semi-field conditions [54][55][56] . ...

CRISPR‐based diagnostics detects invasive insect pests
  • Citing Article
  • October 2023

Molecular Ecology Resources

... All ca. 500 species in the African butterfly tribe Liptenini are suspected to feed on lichens and cyanobacteria (Boyle et al., 2023). Perhaps more surprising are lineages that have moved away from plant feeding entirely. ...

Phylogeny of the Poritiinae (Lepidoptera: Lycaenidae), butterflies with ant associations and unusual lichenivorous diets

Systematic Entomology

... In addition, using just seven landmarks not only reduces the landmark annotation time but also addresses the limitations of GM, specifically for specimens of the order Lepidoptera. The wing margins are prone to damage due to trapping methods, such as recovery from sticky Delta traps or from traps that are not checked daily (12,39,40). Additionally, wing GM in Lepidoptera requires the removal of scales, and the manual cleaning process can further damage the delicate wing margins. ...

A DNA Extraction Method for Insects From Sticky Traps: Targeting a Low Abundance Pest, Phthorimaea absoluta (Lepidoptera: Gelechiidae), in Mixed Species Communities

Journal of Economic Entomology

... Importantly, these clusters of PDF-expressing cells appear conserved across many insect groups, having been identified in locusts, crickets, stick insects and cockroaches [119,120]. There are no previous studies on disco specifically in moths, but publicly available bulk RNA-Seq data from B. mori and Manduca sexta show disco expression in adult in heads and antennae with high larval but minimal pupal expression [65,121]. However, single cell-expression analyses, antibody staining and mRNA in situ hybridization are needed to determine if disco is expressed in or required for the development of clock neurons in Lepidoptera [122]. ...

Population differentiation and structural variation in the Manduca sexta genome across the United States

G3 Genes Genomes Genetics

... We also reconstructed ancestral states of host plant associations and undertook a biogeographical analysis to explore geographical patterns of diversification and the evolution of host plant organ and taxon associations. We integrated phylogenomic and Sanger data for Belidae, sampling all seven tribes and 60% of the extant genera (Combining Sanger sequences data with genomic data for phylogenetic inference has been demonstrated as a feasible approach to resolving deep-level relationships while adding taxa to the phylogeny for tracing the evolutionary history of characters e.g., Zhang et al., 2016;Song et al., 2020;Li et al., 2022a;Li et al., 2022b). We performed ancestral-state reconstruction using the resulting chronogram with stochastic character mapping and event-based likelihood ancestral-area estimation. ...

Phylogeny of gracillariid leaf-mining moths: evolution of larval behaviour inferred from phylogenomic and Sanger data
  • Citing Article
  • October 2021

Cladistics

... To date, 28 complete mitochondrial genomes (mitogenomes) of Triplophysa species have been documented, using mitochondrial genes or complete mitogenome data to explore phylogenetic relationships within the genus 2,[4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] . However, the specific phylogenetic links between T. bombifrons and other species within Triplophysa are still unclear 20,21 . Although sequencing technologies have advanced significantly and reduced sequencing costs, only five chromosome-level genomes from this genus have been sequenced: T. tibetana, T. rosa, T. bleekeri, T. dalaica, and T. siluroides. ...

First Annotated Genome of a Mandibulate Moth, Neomicropteryx cornuta , Generated Using PacBio HiFi Sequencing

Genome Biology and Evolution

... We further analyzed ommatidia distribution across the visual field to approximate parameters of optical performance. Ommatidial counts and diameters were measured using the ommatidia detecting algorithm (ODA; Currea et al., 2021), which measures the number and diameter, D, of corneal lenses by leveraging the periodic nature of the ommatidial lattice through the use of a low-pass filter and the 2D Fourier transform. Corneal lens area, which as opposed to diameter is directly proportional to optical sensitivity, was measured as the area of a circle defined by the facet diameter, A = p (D/2) 2 . ...

Measuring Compound Eye Optics with Microscope and MicroCT Images

... Hence, SVs contribute to the genetic divergence within D. melanogaster. A recent preprint study on Manduca sexta sheds light on the importance of a single inversion which enhances adaptation of the Z chromosome (Mongue and Kawahara, 2020). Generally speaking, SVs attract much less attention than SNPs. ...

Population differentiation and structural variation in the genome of Manduca sexta across the United States