Large-scale mitochondrial DNA analysis of native honey bee Apis mellifera populations reveals a new African subgroup private to the South West Indian Ocean islands

Background: The South West Indian Ocean (SWIO) archipelagos and Madagascar constitute a hotspot of biodiversity with a high rate of endemism. In this area, the endemic subspecies A. m. unicolor has been described in Madagascar. It belongs to the African lineage, one of the four described evolutionary lineages in honey bees. Despite a long beekeeping tradition and several recorded European introductions, few studies have been carried out on the diversity and proportion of honey bee subspecies. In order to identify and define which evolutionary lineages and potential sub-lineages are present in the SWIO, the COI-COII intergenic region and the ND2 gene of the mtDNA were sequenced in honey bee colonies from three archipelagos. An extensive sampling (n = 1184 colonies) was done in the Mascarene (La Réunion, Mauritius, Rodrigues), Seychelles (Mahé, Praslin, La Digue) and Comoros (Grande Comore, Mohéli, Anjouan, Mayotte) archipelagos. Islands genetic diversity was compared to newly sampled populations from Madagascar, continental African and European populations. Results: African lineage haplotypes were found in all islands (except for Rodrigues). Madagascar, Comoros and Seychelles had 100% of A lineage, 95.5% in La Réunion and 56.1% in Mauritius. Among all African colonies detected in the SWIO, 98.1% (n = 633) of COI-COII haplotypes described the presence of the subspecies A. M. unicolor. Both genetic markers revealed i) a new private AI mitochondrial group shared by the SWIO archipelagos and Madagascar distant from continental populations; ii) the private African haplotypes for each island suggested diversity radiation in the archipelagos; iii) the detection of the Comoros archipelago as a possible contact area between insular and continental African populations. The exotic European C and M lineages were only detected in the Mascarene archipelago, but striking differences of proportion were observed among islands. Merely 4.6% of European colonies were found in La Réunion whereas Mauritius cumulated 44%. Here, among the 84 observed COI-COII haplotypes, 50 were newly described including 13 which were private to the SWIO archipelagos and Madagascar. Similarly, 24 of the 34 found ND2 haplotypes were novel which included six haplotypes particular to the SWIO populations. Conclusion: A new African subgroup was described in the SWIO region with mitochondrial genetic evidence that A. m. unicolor is the indigenous subspecies of the archipelagos surrounding Madagascar. (Résumé d'auteur

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. (Résumé d'auteur

Autosomal and mitochondrial adaptation following admixture: a case study on the honeybees of Reunion Island.

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 to samples from Europe, Madagascar, Mauritius, Rodrigues and the Seychelles, revealed the Reunion honeybee population to be composed on 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 one out of the 36 honeybees from Reunion had a mitochondrial genome of European origin, suggesting selection has favoured 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. (Résumé d'auteur

Diversité génétique et phylogéographie de l'abeille Apis mellifera dans les îles du sud-ouest de l'océan Indien

The South West Indian Ocean (SWIO) islands are home to an exceptional flora and fauna and are considered as one the five most important biodiversity hotspots in the world. In most islands of this region, the honeybee Apis mellifera occupies diverse habitats. Regarding its ability as a generalist pollinator, honeybee interacts with native and highly endemic flora. Furthermore, this species is used by human for beekeeping as it is able to produce honey, pollen and other hive products. Within the large group of bees (Apidae), A. mellifera is a model of diversity that has diverged into several lineages and subspecies in its native range. Among the 28 recognized subspecies, A. m. unicolor has been described as endemic to Madagascar and belongs to the African A lineage. The Mascarenes, Comoros and Seychelles archipelagos surround this continental island but the A. mellifera populations present have been little or never studied. The aims of this thesis were to characterize the honeybee from the Mascarenes (La Réunion, Mauritius, Rodrigues), Comoros (Anjouan, Mohéli, Grande Comore, Mayotte) and Seychelles (Mahé, Praslin, La Digue) archipelagos by determining the evolutionary lineages and subspecies present. Secondly, a study of genetic diversity and structure were conducted on these same insular populations. For that, a large sampling was carried (n = 4095 colonies from the SWIO, and 238 from native continental areas) and was combined to molecular analyzes using mitochondrial markers (sequencing of the COI-COII intergenic region and ND2 gene) and nuclear markers (15 microsatellite loci). Three of the four evolutionary lineages (A, C and M) were detected in different proportions in the 10 studied islands. The African A lineage and A. m. unicolor subspecies were predominant in the SWIO excepted for Rodrigues exclusively from the European C lineage. All sampled colonies from the Seychelles and Comoros archipelagos belong to the African lineage while in La Réunion the proportion reach 95.2% and only 54.2% in Mauritius. The presence of the Z African sub-lineage has been described for the first time out of Africa in two Seychelles islands. Moreover, Comoros islands may constitute a contact area between the continental African lineage and A. m. unicolor populations (insular African lineage). The SWIO populations show high levels of nuclear genetic diversity and a structuration by island and archipelago. In addition, SWIO populations strongly differentiated from African and European continental populations. The combined results from different molecular markers favor the hypothesis of an ancient and natural colonization from Madagascar to La Réunion, Mauritius and Seychelles islands. Therefore, the previous referenced interactions between the honeybee and the endemic fauna and flora in the SWIO might be explained by a long cohabitation in addition to its generalist pollinator ability.Les îles du Sud-Ouest de l'océan Indien (SOOI) abritent une faune et une flore exceptionnelle et constituent l'un des cinq hotspot de biodiversité les plus importants au monde. L'abeille domestique Apis mellifera occupe divers habitats dans la majorité de ces îles et interagit avec une flore indigène et endémique. Elle est également exploitée par l'Homme pour l'apiculture. A. mellifera a divergé en plusieurs lignées évolutives et sous-espèces dans son aire d'origine. Parmi-elles, A. m. unicolor a été décrite comme endémique de Madagascar et appartient à la lignée africaine A. Les objectifs de cette thèse étaient de caractériser l'abeille dans les archipels des Mascareignes, Comores et Seychelles en identifiant les lignées évolutives et sous-espèces présentes avec des marqueurs mitochondriaux (région intergénique COI-COII, gène ND2). Dans un second temps, une étude de la diversité et de la structure génétique a été réalisée sur ces mêmes populations insulaires (15 microsatellites). Un total de 4095 colonies ont été échantillonnées dans le SOOI et 238 dans l'aire naturelle continentale. Trois des quatre lignées évolutives (A, C et M) ont été détectées dans les 10 îles étudiées et ce en différentes proportions. La lignée africaine A et A. m. unicolor sont prépondérantes dans le SOOI excepté à Rodrigues (100% lignée européenne C). Dans toutes les îles de l'archipel des Comores et des Seychelles, 100% des colonies échantillonnées appartiennent à la lignée A, 95,2% à La Réunion et seulement 54,2% à Maurice. Les îles de l'archipel des Comores constitueraient une zone de contact entre la lignée africaine continentale et les populations d'A. m. unicolor. La diversité génétique nucléaire est forte dans les archipels du SOOI et est structurée par îles et archipels. En outre, les populations du SOOI se différencient fortement des populations continentales africaines et européennes. La combinaison des différents marqueurs privilégie l'hypothèse d'une colonisation ancienne et naturelle d'A. m. unicolor depuis Madagascar à La Réunion, Maurice et aux Seychelles

Genetic characterization of the honeybee (Apis mellifera) population of Rodrigues Island, based on microsatellite and mitochondrial DNA

International audienceAbstractApis mellifera is present in Rodrigues, an island in the South-West Indian Ocean. The history of the established honeybee population is poorly known, and its biodiversity has never been studied. In this study, maternal origins of A. mellifera in Rodrigues have been assessed with the DraI test and sequencing of the mitochondrial COI-COII region. Nuclear genetic diversity was investigated with 18 microsatellite markers. A total of 524 colonies were sampled from 16 beekeepers in 20 sites. The Rodrigues population was characterized by the absence of the African lineage and presence of three European haplotypes from the C lineage. C1 haplotype was the most frequent (81.3 %), followed by C2 (18.0 %) (characteristics of A. m. ligustica and carnica, respectively) and a new haplotype, C1-Rod (0.7 %). No genetic structure was detected, and genetic diversity was comparable to continental populations. According to approximate Bayesian computation (ABC) analyses, bottleneck scenarios are most likely to have occurred

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

Genetic diversity of the endemic honeybee: Apis mellifera unicolor (Hymenoptera: Apidae) in Madagascar

International audienceAbstract Apis mellifera unicolor is a tropical honeybee endemic of Madagascar. Comprehensive knowledge about its mitochondrial and nuclear genetic diversity and structuration was our main purpose. Samples of worker bees were collected from 867 colonies in 76 sites in Madagascar and 1 reference population in South Africa. PCR-restriction fragment length polymorphism (RFLP) and sequencing were used to reveal variability in the COI–COII mtDNA region. Seventeen microsatellite loci were used for studying the nuclear diversity. Three PCR-RFLP profiles were observed, among which 99.4 % belonged to A1 haplotype, 0.2 % to a new A haplotype, and 0.4 % to A4 haplotype. In microsatellite analysis, moderate genetic diversity values were found for Madagascar, together with low mean number of alleles ranging from 2.47 to 3.88 compared to South Africa. Bayesian clustering assignment methods and principal component analysis (PCA) separated populations into two genetic clusters which matched with geographic areas. Several hypotheses are discussed regarding to the low genetic diversity of A. m. unicolor in its native range