We compared the antennae of two sibling bark beetle species, Dendroctonus rhizophagus Thomas and Bright and Dendroctonus valens LeConte, to identify biologically and taxonomically relevant differences. Specifically, we characterized the diversity of sensilla types by using environmental scanning electron microscopy, quantified the abundance and distribution of the different types of sensilla on the antennal club, and characterized club shape through multivariate and geometric morphometric analyses. We identified four classes of sensilla in both species: chaetica, fluted, trichodea (three distinct types), and basiconica (long and short). ANOVA and MANOVA demonstrated that the number and distribution of sensilla with the potential for chemoreception (i.e., short and long basiconica) differed between species and sexes. Notably, the long sensilla basiconica in the third sensory band were clustered in D. rhizophagus but not in D.valens. Geometric morphometric analysis using 27 landmarks showed that antennal club shape differed significantly between the two species and was not correlated with antennal size. The shape differences were associated with the number and distribution of sensilla.
The bark beetle Dendroctonus rhizophagus is endemic to northwestern Mexico where it kills immature pines < 3 m tall. We report the first investigation of the chemical ecology of this pest of forest regeneration. We used GC-EAD to assess olfactory sensitivity of this species to volatile compounds from: resin of a major host, Pinus arizonica; mid/hindguts of single, gallery-initiating females; and mate-paired males within galleries of attacked host trees in the field. Antennae of both sexes responded to monoterpenes α-pinene, β-pinene and 3-carene as well as to the beetle-derived oxygenated monoterpenes fenchyl alcohol, myrtenal, cis-verbenol, trans-verbenol, verbenone, and myrtenol. These monoterpenes were quantified from pre-emerged D. rhizophagus adults forced to attack host tissue in the laboratory, and from individuals dissected from naturally-attacked hosts at different stages of colonization. In both bioassays, myrtenol and trans-verbenol were the most abundant volatiles, and trans-verbenol was the only one produced in significantly greater quantities by females than males in a naturally-colonized host. Two field experiments were performed to evaluate behavioral responses of D. rhizophagus to antennally-active monoterpenes. Results show that 3-carene was significantly attractive either alone or in a ternary (1:1:1) combination with α-pinene and β-pinene, whereas neither α-pinene nor β-pinene alone were attractive. None of the beetle-associated oxygenated monoterpenes enhanced the attractiveness of the ternary mixture of monoterpenes, while verbenone either alone or combined with the other five oxygenated terpenes reduced D. rhizophagus attraction to the ternary mixture. The results suggest that attraction of D. rhizophagus to the host tree P. arizonica is mediated especially by 3-carene. There was no conclusive evidence for an aggregation or sex attractant pheromone.
Dendroctonus frontalis Zimmermann is considered one of the most important economic and ecological forest pests in the United States, Mexico, and Central America. Recently, two apparent morphological variants of this species were discovered occurring syntopically in Central America and southern Mexico. Morphotype A beetles lack a series of fine parallel ridges on the episternal area of the prothorax that are present on morphotype B. The goal of the present work was to clarify the taxonomic status of the morphotypes of the D. frontalis species complex. Geometric morphometric analyses of seminal rod and spermatheca shape together with the characterization of 16 attributes of external morphology revealed differences in quantitative and qualitative characters that distinguished adults of the two morphotypes from each other as well as from the closely related species Dendroctonus vitei Wood and Dendroctonus mexicanus Hopkins. Karyotype analysis of morphotype B revealed a chromosomal formula (5AA + Xyp) distinct from that found in morphotype A previously reported for D. frontalis (7AA + Xyp). In the laboratory, forced intermorphotype crosses produced F1 progeny but at lower frequency than intramorphotype pairings, and dissections of spermatheca revealed a lower frequency of insemination at least one type of heterotypic cross. Phylogenetic analysis of the D. frontalis species complex based on 786 bp of the cytochrome oxidase I gene indicated that morphotypes B and A are two independent groups with 98% nodal support within D. frontalis. These data provide compelling evidence that the two syntopic morphotypes represent two distinct sibling species.
We provide an illustrated key of species of Dendroctonus Erichson from Mexico and Central America based on characters of the male genitalia and external morphology. The key incorporates newly identified diagnostic characters for this genus that enhance discrimination of particularly difficult sibling species.
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