Anne-Geneviève Bagnères

Biosystematics of Reticulitermes termites in Europe: morphological, chemical and molecular data

Summary: In Europe the most abundant naturally residing termite is the subterranean genus Reticulitermes (Rhinotermitidae). Six phenotypes of Reticulitermes have been identified on the basis of morphological, chemical (cuticular hydrocarbons and soldier defensive secretions), and molecular (enzymatic alleles and mitochondrial ND1 sequence) features. They are R. santonensis in western France, R. grassei in southwestern France, northwestern and southern Spain and Portugal, R. banyulensis in northeastern Spain, central area of the Iberian Peninsula and southwestern France, R. lucifugus in Italy and southeastern France, R. balkanensis in the Balkans and R. sp. nov., a recently identified urban phenotype resembling R. balkanensis, in northern Italy and southeastern France. R. santonensis is close kin to the American species R. flavipes. R. grassei, R. banyulensis and R. lucifugus belong to the same species complex. R. balkanensis and the new phenotype R. sp. nov. are close to R. santonensis regarding cuticular hydrocarbons, to the lucifugus complex regarding DNA and to R. clypeatus from Israel regarding morphology. The species status of these genotypes has been confirmed by the mechanisms of species isolation. Prevention of hybridization depends on the method of colony formation in each species. Swarming dates, differences in pheromones, and infertility prevent hybridization by sexual alates. Interspecific aggression between workers prevents hybridization by neotenics. Behavioral and molecular studies have provided many data on the genetic structure of nests, which varies according to species and location. All colonies of R. santonensis are open all year. The colonies of R. grassei in southern areas and all colonies of R. banyulensis are closed families with generally a single reproductive couple. The colonies of R. grassei in northern areas and the colonies of R. lucifugus are open in the summer and closed in the winter. Based on the here presented data, the taxonomy and the speciation of the Reticulitermes genus in Europe are discussed.

Interspecific recognition among termites of the genusReticulitermes: Evidence for a role for the cuticular hydrocarbons

Two species of termites,Reticulitermes (lucifugus) grassei andR. (l.) banyulensis, show a high degree of aggressivity toward each other. The epicuticular signature, recognized by contact, can be extracted using organic solvents, and the removal of the signature abolished all types of aggressive behavior. The signature can be transferred to lures, where it triggers interspecies aggression. It was found to be mainly present in the apolar fraction of the cuticular extracts, which contained only hydrocarbons, are determined by GC/MS techniques. Chemical recognition contributes towards isolation of the two species belonging to theR. lucifugus complex.

The postpharyngeal glands and the cuticle of Formicidae contain the same characteristic hydrocarbons

Comparison of the contents of the postpharyngeal gland and cuticular hydrocarbons of five species of ant have shown them to contain the same compounds and to be characteristic of the species. For four species (Formica selysi, Camponotus lateralis, Camponotus vagus andManica rubida), quantitative similarity was very close, but more divergent in the fifth (Myrmica rubra). Glands and cuticle ofM. rubra queens were shown to be closely similar to those of workers, except the glands of queens are larger, but the cuticle of larvae was different from that of adult cuticle and postpharyngeal glands.

Cuticular hydrocarbons and defensive compounds of Reticulitermes flavipes (Kollar) and R. santonensis (Feytaud): polymorphism and chemotaxonomy

Colonies ofReticulitermes flavipes andR. santonensis were collected from the southeastern United States (Georgia) and the southwest of France (Charente-maritime). Defensive compounds and cuticular hydrocarbons were identified by gas chromatography-mass spectrometry and quantified by gas chromatography using an internal standard for each caste and all colonies. These analyses show that although the cuticular hydrocarbons ofR. santonensis in Europe andR. flavipes in Georgia are identical, their relative proportions are different. However, the defensive compounds synthesized by their soldiers are different. A strong chemical polymorphism between sympatric colonies ofR. flavipes in the SW United States was detected in terms of both the hydrocarbons of the workers and soldiers and in the defensive secretions of the soldiers. The six defensive secretion phenotypes are based on the presence or absence of terpenes whereas the cuticular hydrocarbon phenotypes are based on significant differences in the proportions of the various components. A multivariate analysis (analysis of principal components) clearly permitted discrimination of four phenotypes (three inR. flavipes and one inR. santonensis) without intermediates. The hydrocarbons responsible for these variations were identified, and it was shown that the variations are neither seasonal nor geographic. The phenotypes of the cuticular hydrocarbons (workers and soldiers) and defensive compounds are linked in each colony, forming in three groups inR. flavipes Georgia, one subdivided into four subgroups according to the defensive secretion phenotypes. The role of these polymorphisms is discussed and ethological tests indicate that the chemical polymorphism do not determine aggressive behavior. The taxonomic significance of these results is considered and two hypothesis are formulated: (1) We only detected a strong genetic polymorphism in one unique species, and we believe thatR. santonensis was introduced into Europe in the last century from oneR. flavipes colony. (2) Chemical variability characterizes the sibling species that can be grouped into the same subspeciesR. flavipes. Unknown mechanisms of reproductive isolation separate them.

A Comparative Genetic Analysis of the Subterranean Termite Genus Reticulitermes (Isoptera: Rhinotermitidae)

Abstract DNA sequencing analysis of the mitochondrial DNA cytochrome oxidase II (COII) region was used to examine genetic variation in the termite genus Reticulitermes Holmgren. We examined 21 species and subspecies from three continents. Sequencing of a 677-bp region of a 780-bp amplicon from 41 individuals and from 17 sequences obtained from GenBank revealed 221 polymorphic sites within the genus. Tajima-Nei distances from species ranged from 0.9 to 12.7%, and parsimony and maximum likelihood analysis revealed several clades within the genus. Reticulitermes flavipes (Kollar) formed a distinct clade along with R. santonensis De Feytaud. European R. lucifugus (Rossi) formed a distinct clade with R. banyulensis (Béziers). Turkish R. lucifugus was distinct relative to European R. lucifugus, and along with R. clypeatus Lash from Israel formed a sister group with R. balkanensis Clément from Greece. This study provides support for the separation of Turkish R. lucifugus from European members of the species. This mitochondrial DNA marker was also able to identify several Reticulitermes specimens from Oklahoma, Texas, Missouri, and South Korea to R. flavipes, R. hageni Banks, R. virginicus (Banks), and R. speratus Shimizu.

Chemical usurpation of a nest by paper wasp parasites

The paper wasp Polistes atrimandibularis is an obligatory social parasite of another Polistes species, P. biglumis bimaculatus. To control the host nest, the parasite sequentially changes the composition of its chemical signature, the cuticular hydrocarbons, during the colonial cycle. Gas chromatography-mass spectrometry of the cuticular hydrocarbons at every stage of the cycle showed that the parasite can switch on and off an entire chemical family, namely, the unsaturated hydrocarbons. In this way the parasite can match the host signature at a critical moment of the colonial cycle.

Cuticular Hydrocarbon Composition Reflects Genetic Relationship Among Colonies of the Introduced Termite Reticulitermes santonensis Feytaud

Nestmate recognition plays a key role in kin selection to maintain colony integrity in social insects. Previous studies have demonstrated that nestmate recognition is dependent on detection of cuticular hydrocarbons. However, the absence of intraspecific aggression between some colonies of Isoptera and social Hymenoptera questions whether kin recognition must occur in social insects. The purpose of this study was to determine if cuticular hydrocarbon similarity and high genetic relatedness could explain the lack of intraspecific aggression among and within colonies of the introduced subterranean termite Reticulitermes santonensis. We performed both GC analysis of cuticular hydrocarbons and genotyping by using 10 DNA microsatellite loci on the same 10 workers from each of 14 parisian colonies. Multivariate analyses demonstrated correspondence between cuticular hydrocarbon patterns and genetic variation. By using a redundancy analysis combining chemical and genetic data, we found that a few hydrocarbons (mainly short vs. long chains; saturated vs. unsaturated alkanes) were associated with most genetic variation. We also found a strong positive correlation between chemical and genetic distances between colonies, thus providing evidence of a genetic basis for cuticular hydrocarbon variation. However, genetic distance did not account for all chemical variation, thus suggesting that some hydrocarbon variation was environmentally derived. Investigation at the intracolony level indicated that cuticular hydrocarbons did not depend on colony social structure. Based on our findings, we speculate that the absence of intraspecific aggression in R. santonensis may result from a loss of diversity in genetically derived recognition compounds in this species that presumably descended from R. flavipes populations imported from North America.

Genetic analysis of the breeding system of an invasive subterranean termite, Reticulitermes santonensis , in urban and natural habitats

Reticulitermes santonensis is a subterranean termite that invades urban areas in France and elsewhere where it causes damage to human‐built structures. We investigated the breeding system, colony and population genetic structure, and mode of dispersal of two French populations of R. santonensis. Termite workers were sampled from 43 and 31 collection points, respectively, from a natural population in west–central France (in and around the island of Oléron) and an urban population (Paris). Ten to 20 workers per collection point were genotyped at nine variable microsatellite loci to determine colony identity and to infer colony breeding structure. There was a total of 26 colonies, some of which were spatially expansive, extending up to 320 linear metres. Altogether, the analysis of genotype distribution, F‐statistics and relatedness coefficients suggested that all colonies were extended families headed by numerous neotenics (nonwinged precocious reproductives) probably descended from pairs of primary (winged) reproductives. Isolation by distance among collection points within two large colonies from both populations suggested spatially separated reproductive centres with restricted movement of workers and neotenics. There was a moderate level of genetic differentiation (FST = 0.10) between the Oléron and Paris populations, and the number of alleles was significantly higher in Oléron than in Paris, as expected if the Paris population went through bottlenecks when it was introduced from western France. We hypothesize that the diverse and flexible breeding systems found in subterranean termites pre‐adapt them to invade new or marginal habitats. Considering that R. santonensis may be an introduced population of the North American species R. flavipes, a breeding system consisting primarily of extended family colonies containing many neotenic reproductives may facilitate human‐mediated spread and establishment of R. santonensis in urban areas with harsh climates.

Genetic evidence for the synonymy of two Reticulitermes species : Reticulitermes flavipes and Reticulitermes santonensis

Abstract By applying the 16S rRNA mitochondrial marker to 434 populations of Reticulitermes termites from North, Central, and South America; France; and Germany with other locations around the world that we have analyzed, identical DNA sequence data were obtained from Reticulitermes flavipes (Kollar) in North America and Germany and for Reticulitermes santonensis Feytaud from undisturbed (nonurban) forested locations in western France. We also discovered identical DNA sequence data from previously unidentified Reticulitermes specimens from South America and Easter Island. Haplotypes F, M, and GG were observed in France; haplotype F in Germany; and haplotype GG was found on Easter Island, Santiago, Chile, and Montevideo, Uruguay. All of these haplotypes are found in numerous states within the continental United States. In light of their well documented morphological, chemical, and phylogenetic relationships, coupled with this new data that directly link these disjunct groups, R. flavipes and R. santonensis should be synonymized. Compared with other studies that largely suggest phyletic similarities, this is the first study that specifically matches haplotypes from North America (where populations of R. flavipes are endemic) with populations in Europe (where R. flavipes, described as R. santonensis, is presumed exotic).

Induced mimicry of colony odors in ants

The cuticular hydrocarbons ofFormica selysi (Formicinae) andMonica rubida (Myrmicinae) reared in single species and in mixed species colonies were determined using gas chromatography (GC) and GC-mass spectrometry. In colonies containing both species, each species modified its species-specific recognition odor. This odor is composed, at least partly, of cuticular hydrocarbons. The cuticular hydrocarbons ofM. rubida consist only of saturated alkanes (n-alkanes and branched alkanes). InF. selysi the mixture also contains unsaturated compounds (monoenes and dienes). In hetero-specific colonies, a new chemical signature developed. This signature resulted from qualitative and quantitative changes in the spectrum of hydrocarbons produced by each species and permitted the two species to inhabit the same nest without displaying interspecific aggression. The readjustment seemed to be more an active synthesis or an active transfer than simply a passive transfer from one species to the other. This may imply that the ants synthesized some components of the hydrocarbon signature of the other species. These synthesizing processes may be activated under particular social environmental conditions.