Paleocene origin of a streamlined digestive symbiosis in leaf beetles.

Timing the acquisition of a beneficial microbe relative to the evolutionary history of its host can shed light on the adaptive impact of a partnership. Here, we investigated the onset and molecular evolution of an obligate symbiosis between Cassidinae leaf beetles and Candidatus Stammera capleta, a γ-proteobacterium. Residing extracellularly within foregut symbiotic organs, Stammera upgrades the digestive physiology of its host by supplementing plant cell wall-degrading enzymes. We observe that Stammera is a shared symbiont across tortoise and hispine beetles that collectively comprise the Cassidinae subfamily, despite differences in their folivorous habits. In contrast to its transcriptional profile during vertical transmission, Stammera elevates the expression of genes encoding digestive enzymes while in the foregut symbiotic organs, matching the nutritional requirements of its host. Despite the widespread distribution of Stammera across Cassidinae beetles, symbiont acquisition during the Paleocene (∼62 mya) did not coincide with the origin of the subfamily. Early diverging lineages lack the symbiont and the specialized organs that house it. Reconstructing the ancestral state of host-beneficial factors revealed that Stammera encoded three digestive enzymes at the onset of symbiosis, including polygalacturonase-a pectinase that is universally shared. Although non-symbiotic cassidines encode polygalacturonase endogenously, their repertoire of plant cell wall-degrading enzymes is more limited compared with symbiotic beetles supplemented with digestive enzymes from Stammera. Highlighting the potential impact of a symbiotic condition and an upgraded metabolic potential, Stammera-harboring beetles exploit a greater variety of plants and are more speciose compared with non-symbiotic members of the Cassidinae.

The leaf beetle Chelymorpha alternans propagates a plant pathogen in exchange for pupal protection.

Many insects rely on microbial protection in the early stages of their development. However, in contrast to symbiont-mediated defense of eggs and young instars, the role of microbes in safeguarding pupae remains relatively unexplored, despite the susceptibility of the immobile stage to antagonistic challenges. Here, we outline the importance of symbiosis in ensuring pupal protection by describing a mutualistic partnership between the ascomycete Fusarium oxysporum and Chelymorpha alternans, a leaf beetle. The symbiont rapidly proliferates at the onset of pupation, extensively and conspicuously coating C. alternans during metamorphosis. The fungus confers defense against predation as symbiont elimination results in reduced pupal survivorship. In exchange, eclosing beetles vector F. oxysporum to their host plants, resulting in a systemic infection. By causing wilt disease, the fungus retained its phytopathogenic capacity in light of its symbiosis with C. alternans. Despite possessing a relatively reduced genome, F. oxysporum encodes metabolic pathways that reflect its dual lifestyle as a plant pathogen and a defensive insect symbiont. These include virulence factors underlying plant colonization, along with mycotoxins that may contribute to the defensive biochemistry of the insect host. Collectively, our findings shed light on a mutualism predicated on pupal protection of an herbivorous beetle in exchange for symbiont dissemination and propagation.

A potential role for overdominance in the maintenance of colour variation in the Neotropical tortoise beetle, Chelymorpha alternans.

The presence of persistent polymorphisms within natural populations elicits the question of how such polymorphisms are maintained. All else equal, genetic drift and natural selection should remove genetic variants from populations. Disassortative mating and overdominance are potential mechanisms for maintaining variation within populations. Here, we consider the potential role of these mechanisms in maintaining variation in colour pattern in the tortoise beetle, Chelymorpha alternans. Five colour morphs distinguished by elytral and pronotal coloration are largely determined by a single locus of large effect with four segregating alleles. As many as four morphs co-occur in natural populations. We first assessed whether disassortative mating might maintain this polymorphism. To test for assortative and disassortative mating, we paired females with two males, one with the same colour pattern as the female and one with a different colour pattern and examined the colour patterns of the offspring. We found strong evidence for random mating as a function of colour pattern. We next assessed whether differences in offspring survival among assortative and disassortative male-female pairs maintain colour variation. Crosses involving disassortative pairings had significantly higher offspring survival during development and resulted in more adult progeny. This result is consistent with the effects of overdominance, whereby outcrossed individuals have higher fitness than their homozygous counterparts. Overall, differences in offspring survival appear to play a greater role in maintaining polymorphisms than nonrandom mating in species.

Symbiont Digestive Range Reflects Host Plant Breadth in Herbivorous Beetles.

Numerous adaptations are gained in light of a symbiotic lifestyle. Here, we investigated the obligate partnership between tortoise leaf beetles (Chrysomelidae: Cassidinae) and their pectinolytic Stammera symbionts to detail how changes to the bacterium's streamlined metabolic range can shape the digestive physiology and ecological opportunity of its herbivorous host. Comparative genomics of 13 Stammera strains revealed high functional conservation, highlighted by the universal presence of polygalacturonase, a primary pectinase targeting nature's most abundant pectic class, homogalacturonan (HG). Despite this conservation, we unexpectedly discovered a disparate distribution for rhamnogalacturonan lyase, a secondary pectinase hydrolyzing the pectic heteropolymer, rhamnogalacturonan I (RG-I). Consistent with the annotation of rhamnogalacturonan lyase in Stammera, cassidines are able to depolymerize RG-I relative to beetles whose symbionts lack the gene. Given the omnipresence of HG and RG-I in foliage, Stammera that encode pectinases targeting both substrates allow their hosts to overcome a greater diversity of plant cell wall polysaccharides and maximize access to the nutritionally rich cytosol. Possibly facilitated by their symbionts' expanded digestive range, cassidines additionally endowed with rhamnogalacturonan lyase appear to utilize a broader diversity of angiosperms than those beetles whose symbionts solely supplement polygalacturonase. Our findings highlight how symbiont metabolic diversity, in concert with host adaptations, may serve as a potential source of evolutionary innovations for herbivorous lineages.

Seasonal and annual abundance of Ephuta wasp (Hymenoptera: Mutillidae) in Panama / Abundancia estacional y anual de la avispa Ephuta (Hymenoptera: Mutillidae) en Panamá

Introduction: Ephuta Say is a widespread New World genus of mutillid wasp whose ecology is poorly understood. Objective: The objective of this study was to determine how Ephuta species abundance varies annually and seasonally driven by weather conditions and hosts. Methods: Ephuta specimens, located in the "old forest" at Barro Colorado Island, were examined from weekly samples (2001-2006) of ten Malaise traps. The monthly abundance of each Ephuta species was compared with monthly average humidity, solar radiation, temperature and rainfall. Results: Nine species and ten morphospecies were identified. Most specimens were collected from March to June. April was the month reported with the greatest abundance. Of the four abiotic variables measured, only average monthly temperature was correlated with Ephuta abundance. Months with the highest number of Ephuta specimens were correlated with suspected host abundance, as indicated by the monthly abundance of Pompilidae (Hymenoptera) captured during the year 2007. Conclusions: We concluded that Ephuta display strong seasonal variation in abundance, with the peaks occurring during the end of the dry season and beginning of the rainy season, which correlate broadly with temperature and the abundance of their pompilid hosts.

Introducción: Ephuta es un género de avispas mutílidas ampliamente distribuido en el Nuevo Mundo y cuya ecología es poco conocida. Objetivo: El objetivo de este trabajo fue determinar cómo varía la abundancia de especies de Ephuta anualmente y estacionalmente debido a las condiciones climáticas y sus hospederos. Métodos: Se examinaron especímenes de Ephuta tomados de muestras semanales (2001-2006) de diez trampas Malaise, ubicadas en el "bosque viejo" de Isla Barro Colorado. La abundancia mensual de cada especie de Ephuta se comparó con el promedio mensual de la humedad, la radiación solar, la temperatura y las precipitaciones. Resultados: Se identificaron nueve especies y diez morfoespecies. La mayoría de los especímenes fueron recolectados de marzo a junio. La mayor abundancia mensual promedio se dio en abril. De las cuatro variables abióticas medidas, solo la temperatura mensual promedio se correlacionó con la abundancia de Ephuta. Los meses con el mayor número de especímenes de Ephuta se correlacionan con la sospechosa abundancia del huésped, como lo indica la abundancia mensual de Pompilidae (Hymenoptera) capturada durante el 2007. Conclusiones: Ephuta muestra una fuerte variación estacional en la abundancia, con picos durante el final de la estación seca y el comienzo de la temporada de lluvias. Esta se correlaciona ampliamente con la temperatura y la abundancia de sus huéspedes pompílidos.

Variation in Host Plant Usage and Diet Breadth Predict Sibling Preference and Performance in the Neotropical Tortoise Beetle Chelymorpha alternans (Coleoptera: Chrysomelidae: Cassidinae).

Specialized interactions between insects and the plants that they consume are one of the most ubiquitous and consequential ecological associations on the plant. Decades of investigation suggest that a narrow diet favors an individual phytophagous insect's performance relative to a dietary generalist. However, this body of research has tended to approach questions of diet breadth and host usage from the perspective of temperate plant-insect associations. Relationships between diet breadth, host usage, and variation in tropical insect preference and performance remain largely uninvestigated. Here we characterize how variation in diet breadth and host usage affect oviposition preference, development, survival, and gain in mass of a Neotropical tortoise beetle Chelymorpha alternans Boheman 1854 (Coleoptera: Chrysomelidae), using a split-brood, sibling experimental design. Host performance was measured after splitting broods among four no-choice host diets. Groups consuming single hosts varied among themselves in developmental time and survival from larva to adult. Performance did not vary among groups consuming multiple and single hosts. Oviposition preference was measured in choice and no-choice tests. Females displayed preference for the original host in both experiments. Developmental time and survival of offspring sourced from the no-choice experiment was measured for two complete generations to explore correlations with female oviposition preference. Preference for the original host correlated with high survivorship and an intermediate developmental time. Survivorship and time to develop were also high on an alternative host that was less preferred. Departures from predictions of prevailing preference-performance hypotheses suggest that host usage presents C. alternans with fitness trade-offs.

Inheritance, distribution and genetic differentiation of a color polymorphism in Panamanian populations of the tortoise beetle, Chelymorpha alternans (Coleoptera: Chrysomelidae).

Intraspecific variation maintained in natural populations has long intrigued scientists and naturalists. One form of this variation, color polymorphisms, provide a rich opportunity to connect genotypic and phenotypic diversity within an ecological and evolutionary context. The existence of color polymorphisms in Panamanian populations of the Neotropical tortoise beetle, Chelymorpha alternans, has been suspected but never systematically explored. To characterize geographic distribution and underlying genetics we sampled a total of 3819 beetles from 28 sites across Panama, quantifying five distinct phenotypes. Two phenotypes, the "metallic" and "rufipennis" are the most widely distributed phenotypes, occurring in nearly all collecting sites. The "veraguensis" phenotype was found to be restricted to the Western end of the Isthmus and the "militaris" phenotypes restricted to sites east of the canal. Controlled matings between phenotypes and reared offspring revealed no indications of reproductive barriers, even among phenotypes which do not co-occur in nature. Color pattern phenotype is largely controlled by Mendelian assortment of four alleles competing at a single locus. A clear dominance hierarchy exists among alleles, with two being co-dominant. Genomic scans from 32 individuals revealed low levels of genetic differentiation, with a small fraction of the genome showing a high degree of divergence. The easily observed variation among populations, simple genetic architecture, and rearing capabilities, make this a promising system for investigating proximate and ultimate factors of phenotypic variation.