Biology subjects

Raguso, R. A.

Publications and source records attributed to Raguso, R. A..

8 recordsLinked to original sources

bioRxiv2026

optix regulates abdominal melanin pigmentation in the tobacco hawkmoth Manduca sexta

Research on butterflies has uncovered a conserved "toolkit" of genes for color pattern development and evolution. One of these genes is optix, a homeobox transcription factor that regulates ommochrome and melanin pigmentation, as well as structural coloration, in nymphalid butterflies. It remains unclear, however, whether optix plays any roles in color patterning outside of the Nymphalidae. We used CRISPR-Cas9 to disrupt optix in the tobacco hornwormmoth Manduca sexta and observed a dramatic abdominal pigmentation phenotype, where orange pigmentation was replaced by black eumelanin. Chemical assays suggest that the orange pigment is not an ommochrome, indicating that optix modulates an alternative, uncharacterized pigment pathway in M. sexta. RNA-seq and chemical analyses of orange and black abdominal scales lead us to speculate that the orange pigment may be a type of melanin, perhaps N-{beta}-alanyldopamine (NBAD) sclerotin. Our results suggest that optix plays a deeply ancestral role in pigment regulation in Lepidoptera, and demonstrates evolutionary flexibility in how it interfaces with pigment chemistry across moths and butterflies.

genetics↗

bioRxiv2025

Squash flowers as microhabitats: the effects of floral temperature and humidity on pollen viability and visitor behavior

Flowers represent intricate microcosms shaped by their chemical and micrometeorological properties. Notable examples include thermogenic flowers that create a warm microclimate for visitors and residents. We document a distinct microclimate within the large flowers of both wild and domesticated squashes (Cucurbita spp.). Unlike thermogenic flowers, squash flower temperatures remain near ambient, but their humidity consistently exceeds ambient levels from bud to senescence, resulting from stomatal and petal transpiration rather than nectar evaporation. Experimentally reducing humidity in greenhouse-grown male squash flowers results in significant pollen tube rupture, directly impacting plant fitness. To explore the role of floral humidity within a broader ecological context, we performed similar humidity manipulations on squash farms to assess impacts on the behavior of their specialist squash bee pollinator (Xenoglossa pruinosa), generalist pollinators (bumblebees, honeybees), and specialist herbivores (cucurbit beetles). Experimentally reduced floral humidity lower visitation frequency by squash bees but have no effect on generalist pollinators. Manipulation of floral humidity did not influence the foraging duration of any pollinators but impacted the residence of male squash bees in wilted flowers compared with unmanipulated flowers. Finally, there was a positive correlation between the dryness of the ambient air and the abundance of squash bees and cucurbit beetles residing in the humid wilted floral chambers. In conclusion, our findings showcase squash flowers as a humid microhabitat that influences reproductive success directly by affecting pollen viability and indirectly by altering interactions with squash bees, their specialist pollinators.

ecology↗

bioRxiv2023

Dying of thirst: Osmoregulation by a hawkmoth pollinator in response to variability in ambient humidity and nectar availability

Climate-induced alterations in flowering phenology can lead to a temporal mismatch between pollinators and the availability of floral resources. Such asynchrony may be especially impactful in desert ecosystems, where flowering time and pollinator emergence are particularly sensitive to rainfall. To investigate the osmoregulation of a desert-living hawkmoth pollinator Manduca sexta, we sampled hemolymph osmolality of over 1000 lab-grown moths held at 20%, 50%, and 80% ambient humidity. Under starvation, the hemolymph osmolality of moths remained within a healthy range from days 1-3, regardless of ambient humidity. However, osmolality levels increase steeply from a baseline of 360-370 mmol/kg to 550 mmol/kg after 4-5 days in low and intermediate humidity and after 5 days in high humidity. Starved moths exposed to low humidity conditions died within 5 days, whereas those in high humidity conditions lived twice as long. Moths fed either synthetic Datura wrightii nectar, synthetic Agave palmeri nectar, or water, maintained osmolality within a healthy baseline range of 350-400 mmol/kg. The same was true for moths that fed on authentic floral nectars. However, moths consumed higher amounts of synthetic nectar, likely due to the non-sugar nectar constituents in the authentic nectar. Finally, simulating a 4-day mismatch between pollinator emergence and nectar availability, we found that a single nectar meal can osmotically rescue moths in dry ambient conditions. Our findings indicate that hemolymph osmolality provides a rapid and accurate biomarker for assessing both the health and relative hydration state of insect pollinators.

physiology↗

bioRxiv2022

Pollen accumulation on hawkmoths varies substantially among moth-pollinated flowers

Using the pollen loads carried by floral visitors to infer their floral visitation behavior is a powerful technique to explore the foraging of wild pollinators. Interpreting these pollen records, however, requires assumptions about the underlying pollen dynamics. To compare visitor foraging across flower species, the most important assumption is that pollen is picked up and retained on the visitor at similar rates. Given differences in pollen presentation traits such as grain number or stickiness even among flowers with similar morphologies, however, the generality of this assumption is unclear. We investigated pollen accumulation on the hawkmoth Manduca sexta, testing the degree to which accumulation differed among flower species and how pollen stickiness affected this accumulation. In no-choice floral visitation assays to six plant species visited by long-tongued hawkmoths in the wild, M. sexta individuals were allowed to visit flowers 1, 2, or 5 times, after which the pollen on their proboscises was removed and counted. We found that the six plant species varied orders of magnitude in the number of pollen grains deposited on the moths, with some placing thousands of grains after a single visit and other placing none after five. Plant species with sticky pollen adhesion mechanisms placed more pollen on the moths and had relatively less pollen accumulation over successive visits than non-sticky plants. Intriguingly, moths carried fewer pollen grains after 5 visits than after 2 visits, suggesting that both sticky and non-sticky pollen was lost during foraging. Together, our results suggest that interpretation of pollen load data should be made cautiously, especially when comparing across plant species.

ecology↗

bioRxiv2022

Floral humidity as a signal--not a cue--in a nocturnal pollination system

Although visual and olfactory floral signals attract pollinators from a distance, at the flowers threshold, pollinators can use floral humidity as an index cue for nectar presence. We evaluate the role of floral humidity in the Datura wrightii-Manduca sexta nocturnal pollination system. In addition to our finding that M. sexta shows strong innate attraction toward humid flowers, we identify the hygrosensing sensillum on their antennae, demonstrate its extreme sensitivity to minute changes in RH, and observe the elimination of moths behavioral preference towards humid flowers following experimental occlusion of the sensilla. Despite Manducas attraction toward humid flowers, we find that floral humidity is not a reliable cue for nectar presence in this system. While Datura floral headspace sustains an enormous humidity gradient, it is not a consequence of nectar evaporation, but an outcome of gas exchange through floral stomata and is decoupled from nectar presence. Using interdisciplinary tools, we demonstrate the function of floral humidity as an attractive signal, not a cue, in this pollination system, thus showcasing an underappreciated modality by which flowers may manipulate their visitors.

ecology↗

bioRxiv2021

Differential gene expression associated with a floral scent polymorphism in the evening primrose Oenothera harringtonii (Onagraceae)

BackgroundPlant volatiles play an important role in both plant-pollinator and plant-herbivore interactions. Intraspecific polymorphisms in volatile production are ubiquitous, but studies that explore underlying differential gene expression are rare. Oenothera harringtonii populations are polymorphic in floral emission of the monoterpene (R)-(-)-linalool; some plants emit (R)-(-)-linalool (linalool+ plants) while others do not (linalool-plants). However, the genes associated with differential production of this floral volatile in Oenothera are unknown. We used RNA-Seq to broadly characterize differential gene expression involved in (R)-(-)-linalool biosynthesis. To identify genes that may be associated with the polymorphism for this trait, we used RNA-Seq to compare gene expression in six different Oenothera harringtonii tissues from each of three linalool+ and linalool-plants. ResultsThree clusters of differentially expressed genes were enriched for terpene synthase activity: two were characterized by tissue-specific upregulation and one by upregulation only in plants with flowers that produce (R)-(-)-linalool. A molecular phylogeny of all terpene synthases identified two putative (R)-(-)-linalool synthase transcripts in Oenothera harringtonii, a single allele of which is found exclusively in linalool+ plants. ConclusionsBy using a naturally occurring polymorphism and comparing different tissues, we were able to identify genes putatively involved in the biosynthesis of (R)-(-)-linalool. Expression of these genes in linalool-plants suggests a regulatory polymorphism, rather than a population-specific loss-of-function allele. Additional terpene biosynthesis-related genes that are up-regulated in plants that emit (R)-(-)-linalool may be associated with herbivore defense, suggesting a potential economy of scale between plant reproduction and defense.

evolutionary biology↗

bioRxiv2020

Emission rates of species-specific volatiles vary across communities of Clarkia species: Evidence for character displacement in floral scent.

A current frontier of character displacement research is to determine if displacement occurs via multiple phenotypic pathways and varies across communities with different species compositions. Here, we conducted the first test for context-dependent character displacement in multimodal floral signals by analyzing variation in floral scent in a system that exhibits character displacement in flower size, and that has multiple types of sympatric communities. In a greenhouse common garden experiment, we measured quantitative variation in volatile emission rates of the progeny of two species of Clarkia from replicated communities that contain one, two, or four Clarkia species. The first two axes of a constrained correspondence analysis, which explained 24 percent of the total variation in floral scent, separated the species and community types, respectively. Of the 23 compounds that were significantly correlated with these axes, nine showed patterns consistent with character displacement. Two compounds produced primarily by C. unguiculata and two compounds produced primarily by C. cylindrica were emitted in higher amounts in sympatry. Character displacement in some volatiles varied across sympatric communities and occurred in parallel with displacement in flower size, demonstrating that this evolutionary process can be context-dependent and may occur through multiple pathways.

evolutionary biology↗

bioRxiv2018

Diversification in the microlepidopteran genus Mompha (Lepidoptera: Gelechioidea: Momphidae) is explained more by tissue specificity than host plant family.

Insect herbivores and their hostplants constitute much of Earths described diversity, but how these often-specialized associations evolve and generate biodiversity is still not fully understood. We combined detailed hostplant data and comparative phylogenetic analyses of the lepidopteran family Momphidae to explore how shifts in the use of hostplant resources, not just hostplant taxonomy, contribute to the diversification of phytophagous insect lineages. We generated two phylogenies primarily from momphid species in the nominate genus, Mompha Hubner. A six-gene phylogeny was constructed with exemplars from Onagraceae hosts in western and southwestern USA and a cytochrome c oxidase subunit 1 (COI) phylogeny utilized both our collected sequences and publicly available accessions from the Barcode of Life Data System. Coalescent-based analyses combined with morphological data revealed ca. 56 undescribed Mompha species-level taxa, many of which are hostplant specialists on southwestern USA Onagraceae. Our phylogenetic reconstructions identified four major momphid clades: 1) an Onagraceae flower- and fruit-feeding clade, 2) a Melastomataceae galling clade, 3) an Onagraceae and Rubiaceae leafmining clade, and 4) a heterogeneous clade associated with multiple hostplant families, plant tissues, and larval feeding modes. Ancestral trait reconstructions on the COI tree identified leafmining on Onagraceae as the ancestral state for Momphidae. Cophylogenetic analyses detected loose phylogenetic tracking of hostplant taxa. Our study finds that shifts along three hostplant resource axes (hostplant taxon, plant tissue type, and larval feeding mode) contributed to the evolutionary success and diversification of Mompha. More transitions between exploited host tissue types than between hostplant families indicated that exploited host tissue (without a change in host) played an unexpectedly large role in the diversification of these moths.

evolutionary biology↗