Johanna Clémencet

Hierarchical analysis of population genetic structure in the monogynous ant Cataglyphis cursor using microsatellite and mitochondrial DNA markers.

Despite having winged queens, female dispersal in the monogynous ant Cataglyphis cursor is likely to be restricted because colonies reproduce by fission. We investigated the pattern of population genetic structure of this species using eight microsatellite markers and a mitochondrial DNA (mtDNA) sequence, in order to examine the extent of female and nuclear gene flow in two types of habitat. Sampling was carried out at a large spatial scale (16 sites from 2.5 to 120 km apart) as well as at a fine spatial scale (two 4.5‐km transects, one in each habitat type). The strong spatial clustering of mtDNA observed at the fine spatial scale strongly supported a restricted effective female dispersal. In agreement, patterns of the mtDNA haplotypes observed at large and fine spatial scales suggested that new sites are colonized by nearby sites. Isolation by distance and significant nuclear genetic structure have been detected at all the spatial scales investigated. The level of local genetic differentiation for mitochondrial marker was 15 times higher than for the nuclear markers, suggesting differences in dispersal pattern between the two sexes. However, male gene flow was not sufficient to prevent significant nuclear genetic differentiation even at short distances (500 m). Isolation‐by‐distance patterns differed between the two habitat types, with a linear decrease of genetic similarities with distance observed only in the more continuous of the two habitats. Finally, despite these low dispersal capacities and the potential use of parthenogenesis to produce new queens, no signs of reduction of nuclear genetic diversity was detected in C. cursor populations.

Habitat-related microgeographic variation of worker size and colony size in the ant Cataglyphis cursor

In social insects, colony size is a crucial life-history trait thought to have major implications for the evolution of social complexity, especially in relation to worker size polymorphism. Yet, little is known about how ecological factors can affect and constrain colony. Here, we explored the pattern of colony-size and worker-size variation in the Mediterranean ant Cataglyphis cursor, in relation to the type of habitats colonized (seaside vs. vineyard). The high level of the water table in the seaside habitat could constrain the depth of C. cursor underground nests and directly constrain its colony size. If worker size increases with colony size, as observed in other ant species, larger colony size and larger workers should be found in the vineyard populations. By comparing worker size among 16 populations, we verified that workers were significantly larger in the vineyard populations. We further determined that the morphological similarities detected among populations from the same habitat type were not due to geographic or genetic proximity. In two populations from each habitat type, the depth of nests was positively correlated with colony size and colony size with worker size. Using a type II regression approach, we further showed that the difference between the two populations in the depth of nest was sufficient to explain the difference in colony size, and similarly, variation in colony size was sufficient to explain variation in worker size. Our results suggest that a single proximate ecological factor could lead to significant variation in major life-history parameters.

Genetic diversity of the honeybee (Apis mellifera L.) populations in the Seychelles archipelago

In the South‐West Indian Ocean, the honeybee Apis mellifera is found on several islands including the Seychelles archipelago. This archipelago is located 1120 km North of Madagascar, where the endemic African subspecies A. m. unicolor occurs. The genetic diversity of the honeybee populations in the Seychelles islands has never been studied, yet this species interacts with highly endemic and indigenous flora. A total of 186 honeybee colonies from the three main islands: Mahé, Praslin and La Digue were collected. In addition, 107 individuals from Madagascar (A. m. unicolor) and 49 from Italy (A. m. ligustica) were analysed as reference populations. The maternal lineages were assessed using PCR‐RFLP (n = 342) and sequencing (n = 121) of the mtDNA COI–COII intergenic region. Intra‐Seychelles nuclear genetic diversity and structure were analysed using 15 microsatellites while comparison with reference populations was done using 14 loci. All Seychellian colonies had mtDNA sequences characteristic of the African evolutionary lineage. Two sub‐lineages were detected: AI sub‐lineage (A1) was dominant (96.7%) on all islands and mostly represented by the subspecies A. m. unicolor, while Z sub‐lineage was observed in six colonies from two islands. No mtDNA characteristic of imported European lineages was detected. Nuclear genetic diversity was high and structured, suggesting restricted gene flow between islands of the archipelago. High nuclear similarities were found among the Seychellian and A. m. unicolor populations, yet significant genetic differentiation was observed. The A. m. ligustica reference population was highly differentiated from the Seychellian honeybee populations.

Assessing hygienic behavior of Apis mellifera unicolor (Hymenoptera: Apidae), the endemic honey bee from Madagascar

Hygienic behavior (HB) is one of the natural mechanisms of honey bee for limiting the spread of brood diseases and Varroa destructor parasitic mite. Objective of our study was to measure HB of Apis mellifera unicolor colonies (N = 403) from three geographic regions (one infested and two free of V. destructor) in Madagascar. The pin-killing method was used for evaluation of the HB. Responses were measured from 3 h 30 min to 7 h after perforation of the cells. Colonies were very effective in detecting perforated cells. In the first 4 h, on average, they detected at least 50% of the pin-killed brood. Six hours after cell perforation, colonies tested (N = 91) showed a wide range of uncapped (0 to 100%) and cleaned cells (0 to 82%). Global distribution of the rate of cleaned cells at 6 h was multimodal and hygienic responses could be split in three classes. Colonies from the three regions showed a significant difference in HB responses. Three hypotheses (geographic, genetic traits, presence of V. destructor) are further discussed to explain variability of HB responses among the regions. Levels of HB efficiency of A. mellifera unicolor colonies are among the greatest levels reported for A. mellifera subspecies. Presence of highly hygienic colonies is a great opportunity for future breeding program in selection for HB.

Kleptoparasitism in the endemic gecko Phelsuma inexpectata : pollen theft from foraging honeybees on Réunion

Interspecific kleptoparasitism (the stealing of already-procured items from other species) has received little attention in tropical reptiles. We report here the second case of kleptoparasitism in tropical lizards, and the first known case of pollen theft by a vertebrate species. Phelsuma inexpectata , a gekkonid lizard endemic to Reunion, was observed robbing pollen pellets from honeybees ( Apis mellifera ) foraging on Latania lontaroides flowers. Video-records were used to obtain reliable information on gecko-honeybee interactions occurring on L. lontaroides inflorescences. During the 19 observation periods (total duration 140 min) a total of 78 attempts were recorded, at least 40% were successful. Both males and females displayed kleptoparasitic behaviour. A high level of gecko kleptoparasitism occurred, with an average one-robbery attempt every 2 min. Behaviour of both the kleptoparasitic P. inexpectata and its host A. mellifera are described.

Genetic diversity and differentiation among insular honey bee populations in the southwest Indian Ocean likely reflect old geographical isolation and modern introductions

With globalization the Western honey bee has become a nearly cosmopolitan species, but it was originally restricted to the Old World. This renowned model of biodiversity has diverged into five evolutionary lineages and several geographic “subspecies.” If Apis mellifera unicolor is indubitably an African subspecies endemic to Madagascar, its relationship with honey bees from three archipelagos in the southwest Indian Ocean (SWIO) hotspot of biodiversity is misunderstood. We compared recent mtDNA diversity data to an original characterization of the nuclear diversity from honey bees in the Mascarenes and Comoros archipelagos, using 14 microsatellites, but also additional mtDNA tRNALeu-cox2 analysis. Our sampling offers the most comprehensive dataset for the SWIO populations with a total of 3,270 colonies from 10 islands compared with 855 samples from Madagascar, 113 from Africa, and 138 from Europe. Comprehensive mitochondrial screening confirmed that honey bees from La Réunion, Mauritius, and Comoros archipelagos are mainly of African origin (88.1% out of 2,746 colonies) and that coexistence with European lineages occurs only in the Mascarenes. PCA, Bayesian, and genetic differentiation analysis showed that African colonies are not significantly distinct on each island, but have diversified among islands and archipelagos. FST levels progressively decreased in significance from European and African continental populations, to SWIO insular and continental populations, and finally among islands from the same archipelago. Among African populations, Madagascar shared a nuclear background with and was most closely related to SWIO island populations (except Rodrigues). Only Mauritius Island presented clear cytoplasmic disequilibrium and genetic structure characteristic of an admixed population undergoing hybridization, in this case, between A. m. unicolor and A. m. ligustica, A. m. carnica and A. m. mellifera-like individuals. Finally, global genetic clustering analysis helped to better depict the colonization and introduction pattern of honey bee populations in these archipelagos.

Pollen morphology of melliferous plants for Apis mellifera unicolor in the tropical rainforest of Ranomafana National Park, Madagascar

ABSTRACT The morphological diversity of pollen brings important information on the taxonomy of plants present in past and current ecosystems. Regarding the rich biodiversity and the large number of endemic plant species on the island of Madagascar, pollen reference collections are still scarce for this part of the world. The main objective of this study was to conduct a preliminary investigation of pollen morphology of flowering plants visited by honeybees in a tropical rainforest of the south-east part of Madagascar during 2014 to 2015 (Ranomafana National Park – Fianarantsoa). Samples of 135 species belonging to 105 genera and 52 families were processed, employing the standard acetolysis method. Pollen reference slides of these melliferous plants were mounted. These pollen grains were then photographed and measured using light microscopy. The 135 collected species include 54 trees, 47 shrubs, 21 herbs, 12 lianas and one epiphyte, among which 85 taxa are endemic, 46 indigenous and four exotic. Pollen morphological characteristics of 106 species (85% endemic) are described here for the first time. Since these endemic plant taxa have co-evolved with the endemic honeybee of Madagascar, Apis mellifera unicolor, the implications of the pollen grain morphology for plant–pollinator co-evolution are discussed.

Autosomal and Mitochondrial Adaptation Following Admixture: A Case Study on the Honeybees of Reunion Island

Abstract The honeybee population of the tropical Reunion Island is a genetic admixture of the Apis mellifera unicolor subspecies, originally described in Madagascar, and of European subspecies, mainly A. m. carnica and A. m. ligustica, regularly imported to the island since the late 19th century. We took advantage of this population to study genetic admixing of the tropical-adapted indigenous and temperate-adapted European genetic backgrounds. Whole genome sequencing of 30 workers and 6 males from Reunion, compared with samples from Europe, Madagascar, Mauritius, Rodrigues, and the Seychelles, revealed the Reunion honeybee population to be composed on an average of 53.2 ± 5.9% A. m. unicolor nuclear genomic background, the rest being mainly composed of A. m. carnica and to a lesser extent A. m. ligustica. In striking contrast to this, only 1 out of the 36 honeybees from Reunion had a mitochondrial genome of European origin, suggesting selection has favored the A. m. unicolor mitotype, which is possibly better adapted to the island’s bioclimate. Local ancestry was determined along the chromosomes for all Reunion samples, and a test for preferential selection for the A. m. unicolor or European background revealed 15 regions significantly associated with the A. m. unicolor lineage and 9 regions with the European lineage. Our results provide insights into the long-term consequences of introducing exotic specimen on the nuclear and mitochondrial genomes of locally adapted populations.

Genetic diversity of Varroa destructor parasitizing Apis mellifera unicolor in Madagascar

Varroadestructor is an invasive alien species that has been reported parasitizing the endemic honey bee of Madagascar, Apis mellifera unicolor, since 2010. Studying its nuclear genetic diversity and structure was our main goal. Using 11 microsatellite loci and 344 mites collected from 12 apiaries, we observed a low genetic diversity, with only 8 multilocus genotypes (MLG) identified. V. destructor populations form a single genetic cluster, clonal richness ranged from 0.02 to 0.20, and number of MLG within apiaries varied between one and six MLGs. About 69.5% of the mites analyzed harbored the same genotype (100%, homozygous), and 23.3% had a genotype differing by a single allele. The overall low diversity observed suggests one or multiple introductions of similar genotypes. The greater abundance of MLGs in High Land apiaries (eight MLGs) than on the east coast (two MLGs) and the presence of particular MLGs in High Land apiaries favor the hypothesis that V. destructor has been first introduced close to the international airport, and then spread to other regions by commercial exchanges.