Huai-Jun Xue

Male mate recognition via cuticular hydrocarbons facilitates sexual isolation between sympatric leaf beetle sister species

Chemical signals in insects have been documented to play an important role in mate recognition, and divergence in chemical signals can often cause sexual isolation between closely related species or populations within species. We investigated the role of cuticular hydrocarbons (CHCs), short distance chemical signals, in male mate recognition between the two sympatric elm leaf beetles, Pyrrhalta maculicollis and Pyrrhaltaaenescens. Mating experiments demonstrated that strong sexual isolation between the two species was driven by CHCs divergence. Males preferred to mate with conspecific females with intact conspecific CHCs or conspecific CHCs reapplied after removal. Males also preferred heterospecific females that were treated with conspecific CHCs. Chemical analysis showed that the CHC profiles differ significantly between species. In P. maculicollis dimethyl-branched alkanes between C29 and C35 account for the majority of the saturated alkanes while the CHC profile of P. aenescens mostly consisted of monomethyl-branched alkanes between C22 and C29. Additionally, some compounds, such as 12,18-diMeC32, 12,18-diMeC34, are unique to P. maculicollis.

Development of a positive preference-performance relationship in an oligophagous beetle: adaptive learning?

The relationship between oviposition preference and larval performance is a central topic in insect–plant biology. In this study, we investigate whether the oligophagous flea beetle, Altica fragariae Nakane (Coleoptera: Chrysomelidae), exhibits a positive preference–performance relationship, and whether oviposition preference develops over time. We tested the beetles using four sympatric plant species: Duchesnea indica (Andrews) Focke (the normal host plant), Agrimonia pilosa Ledeb. (a secondary host plant), and Potentilla chinensis Ser. and Sanguisorba officinalis L. (host plants of two related Altica species) (all Rosaceae). In no‐choice experiments, both oviposition rate and offspring fitness parameters (eclosion rate, development time, and body mass) were highest on D. indica. Oviposition rate was much lower on P. chinensis than on A. pilosa, whereas offspring fitness parameters did not differ significantly between these two host plants. Offspring fitness were lowest for S. officinalis, and adult females refused to oviposit on this acceptable non‐host in a no‐choice situation. Repeated two‐choice experiments showed that the proportion of oviposition on one of the novel host plants decreased significantly over time when the alternative host plant was D. indica. In repeated two‐choice experiments using A. pilosa and P. chinensis, females mainly fed on A. pilosa but distributed their eggs equally over the two host plants, in accordance with the lack of difference in offspring fitness on those hosts. Together, these results showed that A. fragariae females develop a positive preference–performance relationship over time. We suggest that A. fragariae achieves this through adaptive learning of oviposition preference: not only does the female learn to discriminate among the host plants when there is a fitness difference for her offspring, but the female also fails to discriminate when there is no fitness difference.

Distinct species or colour polymorphism? Life history, morphology and sequence data separate two Pyrrhalta elm beetles (Coleoptera: Chrysomelidae)

The elm leaf beetles Pyrrhalta maculicollis (Motschulsky, 1853) and P. aenescens (Fairmaire, 1878) (Chrysomelidae: Galerucinae) are considered to be two separate species that occur in sympatry and synchrony on the same plant host. Conventionally they are distinguished by differences in elytron and pronotum colour, but this variation could just represent intra-specific polymorphism. To test the status of P. maculicollis and P. aenescens we compared their biology, larval and adult morphological characters, and molecular genetics. Using laboratory-bred cohorts of either colour type, biological and morphological analyses showed great similarities in life cycle, body size, external morphology of eggs and pupae, and the presence of spines on the internal sac of the aedeagus not seen in other species of Pyrrhalta. However, both groups showed consistent differences in colour patterns, tubercles and setae of the larvae, while the number of spines on the internal sac also differed consistently. The molecular phylogenetic analysis based on mitochondrial (COI, COII) and nuclear (ITS2) markers recovered both colour types as deeply separated, reciprocally monophyletic lineages, and in addition found P. maculicollis to be split into two divergent subgroups by both markers. A time-calibrated tree that included several further species of Pyrrhalta and the related Galerucella indicated that P. maculicollis and P. aenescens separated in the Miocene, at around 7.5 Ma (95% CI, 9.5–5.6 Ma).

Reproductive barriers between two sympatric beetle species specialized on different host plants

Knowledge on interspecific pre‐ and post‐zygotic isolation mechanisms provides insights into speciation patterns. Using crosses (F1 and backcrosses) of two closely related flea beetles species, Altica fragariae and A. viridicyanea, specialized on different hosts in sympatry, we measured: (a) the type of reproductive isolation and (b) the inheritance mode of preference and host‐specific performance, using a joint‐scaling test. Each species preferred almost exclusively its host plant, creating strong prezygotic isolation between them, and suggesting that speciation may occur at least partly in sympatry. Reproductive isolation was intrinsic between females of A. fragariae and either A. viridicyanea or F1 males, whereas the other crosses showed ecologically dependent reproductive isolation, suggesting ecological speciation. The genetic basis of preference and performance was at least partially independent, and several loci coded for preference, which limits the possibility of sympatric speciation. Hence, both ecological and intrinsic factors may contribute to speciation between these species.

Genetic analysis of feeding preference in two related species of Altica (Coleoptera: Chrysomelidae: Alticinae)

Abstract 1. Genetic analysis of feeding preference can make an important contribution to our understanding of the evolution of host‐plant selection in phytophagous insects. Two closely related flea beetles, Altica viridicyanea (Baly) and Altica fragariae Nakane, with separate host plants [Geranium wilfordii Maxim. and Duchesnea indica (Andrews) Focke respectively] were hybridised to analyse the inheritance mode of feeding preference.

Host Plant Use in Sympatric Closely Related Flea Beetles

Abstract Studies on strategies of host plant use in sympatric-related species are significant to the theory of sympatric speciation. Altica fragariae Nakane and Altica koreana Ogloblin are sympatric closely related flea beetles found in Beijing, northern China. All their recorded host plants are in the subfamily Rosoideae of the Rosaceae, so we regard them as a model system to study interactions between herbivorous insects and plant-insect co-evolution. We conducted a set of experiments on the host preference and performance of these flea beetles to study whether these closely related species have the ability to use sympatric novel host plants and whether monophagous and oligophagous flea beetles use the same strategy in host plant use. Oviposition preference experiments showed that A. koreana, a monophagous flea beetle, displayed high host fidelity. However, A. fragariae, which is oligophagous, often made “oviposition mistakes,” ovipositing on nonhost plants such as Potentilla chinensis, the host plant of A. koreana, although normal host plants were preferred over novel ones. Larval performance studies suggested that A. fragariae was able to develop successfully on P. chinensis. Feeding experiences of larvae had no effect on feeding preference, oviposition preference, and fecundity of adults. However, females were impaired in their reproductive ability when fed on nonhost plants. Therefore, A. fragariae finished their development of larval stages on P. chinensis and came back to their primary host plant, Duchesnea indica, for feeding and reproduction after eclosion.

Assortative mating between two sympatric closely-related specialists: inferred from molecular phylogenetic analysis and behavioral data

Host plant shifting of phytophagous insects can lead to the formation of host associated differentiation and ultimately speciation. In some cases, host plant specificity alone acts as a nearly complete pre-mating isolating barrier among insect populations. We here test whether effective pre-mating isolation and host-independent behavioral isolation have evolved under the condition of extreme host specilization using two sympatric flea beetles with incomplete post-mating isolation under laboratory conditions. Phylogenetic analysis and coalescent simulation results showed that there is a limited interspecific gene flow, indicating effctive isolation between these species. Three types of mating tests in the absence of host plant cues showed that strong host-independent behavioral isolation has evolved between them. We conclude that almost perfect assortative mating between these two extreme host specialists results from a combination of reduced encounter rates due to differential host preference and strong sexual isolation.

Recent Speciation in Three Closely Related Sympatric Specialists: Inferences Using Multi-Locus Sequence, Post-Mating Isolation and Endosymbiont Data

Shifting between unrelated host plants is relatively rare for phytophagous insects, and distinct host specificity may play crucial roles in reproductive isolation. However, the isolation status and the relationship between parental divergence and post-mating isolation among closely related sympatric specialists are still poorly understood. Here, multi-locus sequence were used to estimate the relationship among three host plant–specific closely related flea beetles, Altica cirsicola, A. fragariae and A. viridicyanea (abbreviated as AC, AF and AV respectively). The tree topologies were inconsistent using different gene or different combinations of gene fragments. The relationship of AF+(AC+AV) was supported, however, by both gene tree and species tree based on concatenated data. Post-mating reproductive data on the results of crossing these three species are best interpreted in the light of a well established phylogeny. Nuclear-induced but not Wolbachia-induced unidirectional cytoplasmic incompatibility, which was detected in AC-AF and AF-AV but not in AC-AV, may also suggest more close genetic affinity between AC and AV. Prevalence of Wolbachia in these three beetles, and the endosymbiont in most individuals of AV and AC sharing a same wsp haplotype may give another evidence of AF+(AC+AV). Our study also suggested that these three flea beetles diverged in a relative short time (0.94 My), which may be the result of shifting between unrelated host plants and distinct host specificity. Incomplete post-mating isolation while almost complete lineage sorting indicated that effective pre-mating isolation among these three species should have evolved.

Complete mitochondrial genome of Spiniphilus spinicornis (Coleoptera: Vesperidae: Philinae) and phylogenetic analysis among Cerambycoidea.

Abstract Spiniphilus spinicornis belongs to subfamily Philinae of family Vesperidae from cerambycoid assemblage. The first complete mitogenome of Spiniphilus spinicornis was reported. The genome is 15 707 bp in length and contains the typical 37 genes that are arranged in the same order as that of the putative ancestor of beetles. The total base composition of the mitogenome is 30.9% for A, 11.1% for C, 19.3% for G, and 38.6% for T. The genome organization, nucleotide composition, and codon usage do not differ significantly from other martens. The percentage of A + T is 69.5%. The first complete mitogenome of subfamily Philinae could be used in studies of molecular systematics, phylogenetic, and conservation genetics. Phylogenetic analysis showed that Cerambycoidea was monophyly with high support value. Lamiinae, Cerambycinae, and Philinae were monophyly too, respectively.

Endogenous cellulolytic enzyme systems in the longhorn beetle Mesosa myops (Insecta: Coleoptera) studied by transcriptomic analysis

The Cerambycidae (longhorn beetle) is a large family of Coleoptera with xylophagous feeding habits. Cellulose digestion plays an important role in these wood-feeding insects. In this study, transcriptomic technology was used to obtain one glycoside hydrolase family 45 (GH45) cellulase and seven GH5 cellulases from Mesosa myops, a typical longhorn beetle. Analyses of expression dynamics and evolutionary relationships provided a complete description of the cellulolytic system. The expression dynamics related to individual development indicated that endogenous GH45 and GH5 cellulases dominate cellulose digestion in M. myops. Evolutionary analyses suggested that GH45 cellulase gene is a general gene in the Coleoptera Suborder Polyphaga. Evolutionary analyses also indicated that the GH5 cellulase group in Lamiinae longhorn beetles is closely associated with wood feeding. This study demonstrated that there is a complex endogenous cellulolytic system in M. myops that is dominated by cellulases belonging to two glycoside hydrolase families.