A cure for the blues: opsin duplication and subfunctionalization for short-wavelength sensitivity in jewel beetles (Coleoptera: Buprestidae).

BACKGROUND: Arthropods have received much attention as a model for studying opsin evolution in invertebrates. Yet, relatively few studies have investigated the diversity of opsin proteins that underlie spectral sensitivity of the visual pigments within the diverse beetles (Insecta: Coleoptera). Previous work has demonstrated that beetles appear to lack the short-wavelength-sensitive (SWS) opsin class that typically confers sensitivity to the "blue" region of the light spectrum. However, this is contrary to established physiological data in a number of Coleoptera. To explore potential adaptations at the molecular level that may compensate for the loss of the SWS opsin, we carried out an exploration of the opsin proteins within a group of beetles (Buprestidae) where short-wave sensitivity has been demonstrated. RNA-seq data were generated to identify opsin proteins from nine taxa comprising six buprestid species (including three male/female pairs) across four subfamilies. Structural analyses of recovered opsins were conducted and compared to opsin sequences in other insects across the main opsin classes-ultraviolet, short-wavelength, and long-wavelength. RESULTS: All nine buprestids were found to express two opsin copies in each of the ultraviolet and long-wavelength classes, contrary to the single copies recovered in all other molecular studies of adult beetle opsin expression. No SWS opsin class was recovered. Furthermore, the male Agrilus planipennis (emerald ash borer-EAB) expressed a third LWS opsin at low levels that is presumed to be a larval copy. Subsequent homology and structural analyses identified multiple amino acid substitutions in the UVS and LWS copies that could confer short-wavelength sensitivity. CONCLUSIONS: This work is the first to compare expressed opsin genes against known electrophysiological data that demonstrate multiple peak sensitivities in Coleoptera. We report the first instance of opsin duplication in adult beetles, which occurs in both the UVS and LWS opsin classes. Through structural comparisons of known insect opsins, we suggest that opsin duplication and amino acid variation within the chromophore binding pocket explains sensitivity in the short-wavelength portion of the visible light spectrum in these species. These findings are the first to reveal molecular complexity of the color vision system within beetles.

Differences in spectral selectivity between stages of visually guided mating approaches in a buprestid beetle.

Spectral mating preferences were examined in male Agrilus angustulus (Buprestidae: Coleoptera), a member of a taxon known for its high species diversity and striking metallic coloration. The spectral emission profile of a typical A. angustulus female displays low chroma, broadly overlapping that of the green oak leaves they feed and rest upon, while also including longer wavelengths. To pinpoint behaviorally significant spectral regions for A. angustulus males during mate selection, we observed their field approaches to females of five Agrilus planipennis color morphs that have greater chroma than the normal conspecific female targets. Agrilus angustulus males would initially fly equally frequently toward any of the three longest wavelength morphs (green, copper and red) whose spectral emission profiles all overlap that of typical A. angustulus females. However, they usually only completed approaches toward the two longest wavelength morphs, but not the green morphs. Thus, spectral preference influenced mate selection by A. angustulus males, and their discrimination of suitable targets became greater as these targets were approached. This increasing spectral discrimination when approaching targets may have evolved to allow female emissions to remain somewhat cryptic, while also being visible to conspecifics as distinct from the background vegetation and heterospecific competitors.

Effects of ambient temperature on egg and larval development of the invasive emerald ash borer (Coleoptera: Buprestidae): implications for laboratory rearing.

The emerald ash borer, Agrilus planipennis Fairmaire, an invasive beetle from Asia causing large scale ash (Fraxinus) mortality in North America, has been extremely difficult to rear in the laboratory because of its long life cycle and cryptic nature of immature stages. This lack of effective laboratory-rearing methods has not only hindered research into its biology and ecology, but also mass production of natural enemies for biological control of this invasive pest. Using sticks from the alternate host plant, Fraxinus uhdei (Wenzig) Lingelsh, we characterized the stage-specific development time and growth rate of both emerald ash borer eggs and larvae at different constant temperatures (12-35 degrees C) for the purpose of developing effective laboratory-rearing methods. Results from our study showed that the median time for egg hatching decreased from 20 d at 20 degrees C to 7 d at 35 degrees C, while no emerald ash borer eggs hatched at 12 degrees C. The developmental time for 50% of emerald ash borer larvae advancing to third, fourth, and J-larval stages at 20 degrees C were 8.3, 9.1, and 12.3 wk, respectively, approximately two times longer than at 30 degrees C for the corresponding instars or stages. In contrast to 30 degrees C, however, the development times of emerald ash borer larvae advancing to later instars (from oviposition) were significantly increased at 35 degrees C, indicating adverse effects of this high temperature. The optimal range of ambient temperature to rear emerald ash borer larvae should be between 25-30 degrees C; however, faster rate of egg and larval development should be expected as temperature increases within this range.

Establishment and abundance of Tetrastichus planipennisi (Hymenoptera: Eulophidae) in Michigan: potential for success in classical biocontrol of the invasive emerald ash borer (Coleoptera: Buprestidae).

Tetrastichus planipennisi Yang is a gregarious larval endoparasitoid native to China and has been introduced to the United States since 2007 for classical biological control of the invasive emerald ash borer, Agrilus planipennis Fairmaire, an exotic beetle responsible for widespread ash mortality. Between 2007-2010, T. planipennisi adults (3,311-4,597 females and approximately 1,500 males per site) were released into each of six forest sites in three counties (Ingham, Gratiot, and Shiawassee) of southern Michigan. By the fall of 2012, the proportion of sampled trees with one or more broods of T. planipennisi increased to 92 and 83% in the parasitoid-release and control plots, respectively, from 33 and 4% in the first year after parasitoid releases (2009 fall for Ingham county sites and 2010 for other sites). Similarly, the mean number of T. planipennisi broods observed from sampled trees increased from less than one brood per tree in the first year after parasitoid releases to 2.46 (at control plots) to 3.08 (at release plots) broods by the fall of 2012. The rates of emerald ash borer larval parasitism by T. planipennisi also increased from 1.2% in the first year after parasitoid releases to 21.2% in the parasitoid-release plots, and from 0.2 to 12.8% for the control plots by the fall of 2012. These results demonstrate that T. planipennisi is established in southern Michigan and that its populations are increasing and expanding. This suggests that T. planipennisi will likely play a critical role in suppressing emerald ash borer populations in Michigan.

Male-produced pheromone of Spathius agrili, a parasitoid introduced for the biological control of the invasive emerald ash borer, Agrilus planipennis.

The braconid wasp, Spathius agrili, has been released in the U.S. as a biocontrol agent for the invasive emerald ash borer (Coleoptera: Buprestidae: Agrilus planipennis), a destructive pest of ash trees (Fraxinus spp.). We identified and synthesized seven male-specific volatile compounds. Three of these, dodecanal, (4R,11E)-tetradecen-4-olide, and (Z)-10-heptadecen-2-one, were the key behaviorally active components in flight tunnel bioassays. Male specificity was demonstrated by gas chromatographic comparison of male and female volatile emissions and whole body extracts. Identifications were aided by coupled gas chromatographic-mass spectrometric (GC-MS) analysis, microchemical reactions, NMR, chiral GC analysis, and GC and MS comparison with authentic standards. Both the racemic and chiral forms of the γ-lactone, as well as both E- and Z-isomers were synthesized. Flight tunnel behavioral tests showed positive male and female S. agrili responses to both natural pheromone and synthetic blends, with upwind flight and landing on the source. Large field-cage tests, using yellow sticky traps baited with pheromone, captured approximately 50% of the released male and female wasps in 24-h periods. The use of pheromone-baited traps in the field could simplify the current detection method for determining parasitoid establishment (i.e., laboriously felling and peeling ash trees for recovery of S. agrili from infested EAB larvae).

Behavioral evidence for a contact sex pheromone component of the emerald ash borer, Agrilus planipennis Fairmaire.

The cuticular hydrocarbon profiles of emerald ash borers, Agrilus planipennis, were examined to determine if there are differences in these compounds between the sexes. We also assessed feral male EAB in the field for behavioral changes based on the application of a female-specific compound to dead, solvent-washed beetles. Males in the field spent significantly more time attempting copulation with dead, pinned female beetles coated with a three-beetle-equivalent dose of 3-methyltricosane than with solvent-washed beetles or those coated in 3-methyltricosane at lower concentrations. Males in the field spent the most time investigating pinned dead, unwashed female beetles. In the laboratory, sexually mature males were presented with one of several mixtures applied in hexane to filter paper disks or to the elytra of dead female beetles first washed in solvent. Male EAB also spent more time investigating dead beetles treated with solution applications that contained 3-methyltricosane than dead beetles and filter paper disks treated with male body wash or a straight-chain hydrocarbon not found on the cuticle of EAB.

Complicity or conflict over sexual cannibalism? Male risk taking in the praying mantis Tenodera aridifolia sinensis.

Male complicity versus conflict over sexual cannibalism in mantids remains extremely controversial, yet few studies have attempted to establish a causal relationship between risk of cannibalism and male reproductive behavior. We studied male risk-taking behavior in the praying mantid Tenodera aridifolia sinensis by altering the risk imposed by females and measuring changes in male behavior. We show that males were less likely to approach hungrier, more rapacious females, and when they did approach, they moved more slowly, courted with greater intensity, and mounted from a greater distance. Similarly, when forced to approach females head-on, within better view and better reach of females, males also approached more slowly and courted with greater intensity. Thus, males behaved in a manner clearly indicative of risk avoidance, and we support the hypothesis of sexual conflict over sexual cannibalism.

Factors affecting the flight capacity of Tetrastichus planipennisi (Hymenoptera: Eulophidae), a classical biological control agent of Agrilus planipennis (Coleoptera: Buprestidae).

The dispersal characteristics of a biological control agent can have direct implications on the ability of that agent to control populations of a target host. Tetrastichus planipennisi Yang (Hymenoptera: Eulophidae) is a parasitic wasp native to eastern Asia that has been introduced into the United States as part of a classical biological control program against the emerald ash borer Agrilus planipennis Fairmaire (Coleoptera: Buprestidae). We used computer-monitored flight mills to investigate the role of age, feeding status, mating status, and size on the flight capacity of female T. planipennisi over a 24-h period. We also compared flight capacity between sexes. Flight distance of female T. planipennisi representative of populations released in the biological control program averaged 1.26 km in 24 h with a maximum flight of just over 7 km. Median flight distance, however, was 422 m. The flight capacity of females fed a honey-water solution was 41× that of females provided only water, who flew very little. Larger females were capable of flying farther distances, but age did not affect the flight capacity of females up to 70 d posteclosion. Females dispersed 6× farther than did their smaller, male counterparts. The implications of our findings to host-parasitoid interactions and release protocols for distributing T. planipennisi are discussed.

Effects of host plant and larval density on intraspecific competition in larvae of the emerald ash borer (Coleoptera: Buprestidae).

Competition for food, mates, and space among different individuals of the same insect species can affect density-dependent regulation of insect abundance or population dynamics. The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is a serious invasive pest of North American ash (Fraxinus spp.) trees, with its larvae feeding in serpentine galleries between the interface of sapwood and phloem tissues of ash trees. Using artificial infestation of freshly cut logs of green ash (Fraxinus pennsylvanica Marshall) and tropical ash (Fraxinus uhdei [Wenzig] Lingelsh) with a series of egg densities, we evaluated the mechanism and outcome of intraspecific competition in larvae of A. planipennis in relation to larval density and host plant species. Results from our study showed that as the egg densities on each log (1.5-6.5 cm in diameter and 22-25 cm in length) increased from 200 to 1,600 eggs per square meter of surface area, larval survivorship declined from ≍68 to 10% for the green ash logs, and 86 to 55% for tropical ash logs. Accordingly, larval mortality resulting from cannibalism, starvation, or both, significantly increased as egg density increased, and the biomass of surviving larvae significantly decreased on both ash species. When larval density was adjusted to the same level, however, larval mortality from intraspecific competition was significantly higher and mean biomasses of surviving larvae was significantly lower in green ash than in tropical ash. The role of intraspecific competition of A. planipennis larvae in density-dependent regulation of its natural population dynamics is discussed.

Interspecific pheromone plume interference among sympatric heliothine moths: a wind tunnel test using live, calling females.

Three species of North American heliothine moths were used to determine the level at which interspecific female interference of male attraction to conspecific females occurs. We used live calling females of Heliothis virescens, H. subflexa, and Helicoverpa zea, as lures for conspecific males in a wind tunnel, and then placed heterospecific females on either side of the original species such that the plumes of the three females overlapped downwind. In nearly all combinations, in the presence of heterospecific females, fewer males flew upwind and contacted or courted the source than when only conspecific females were used in the same spatial arrangement. Males did not initiate upwind flight to solely heterospecific female arrangements. Our results show that the naturally emitted pheromone plumes from heterospecific females of these three species can interfere with the ability of females to attract conspecific males when multiple females are in close proximity. However, the fact that some males still located their calling, conspecific females attests to the ability of these male moths to discriminate point source odors by processing the conflicting information from interleaved strands of attractive and antagonistic odor filaments on a split-second basis.