Unveiling introgression in bumblebee (Bombus terrestris) populations through mitogenome-based markers

The bumblebee, Bombus terrestris, is an important pollinator commercially used on a global scale. The exported subspecies B. t. terrestris has colonised diverse environments, in some cases displacing wild pollinators to the verge of local extinction. In this sense, the native Iberian subspecies B. t. lusitanicus may be threatened by the subspecies B. t. terrestris, naturally distributed from the Pyrenees to Central Europe but also observed in southern Spain due to escapes from commercial nests. Mitochondrial genomes have a low recombination rate and a small effective population size owing to their maternal inheritance, thus providing an accurate approach to study hybridisation events between populations. Therefore, we present the sequences of the mitogenomes of both subspecies as a molecular framework to select suitable markers to detect possible introgression events between them. We used metagenomics to obtain approximately 17 kbp of the mitogenome from both subspecies. Their mitogenomes differed in 358 bp (excluding the AT-rich region). Four mitogenomic fragments were selected to be tested as subspecific diagnostic markers. A RFLP detected in the gene nad2 (NADH dehydrogenase subunit 2) has proven to be an efficient, quick and cost-effective tool to assess the dispersion of the non-endemic subspecies into Iberian native populations. Subspecific haplotypes were observed in both morphological subspecies, suggesting introgression events in the northern natural contact area and in the new human-mediated contact area in the south of the Iberian Peninsula.     

Is the exotic bumblebee Bombus terrestris really invading Tasmanian native vegetation?

There has been a great deal of disagreement surrounding the capacity of Bombus terrestris to invade Tasmanian native vegetation. This paper reviews the conflicting findings of previous surveys of the invasion of Tasmania by B. terrestris, and presents new data from the 2004–2005 austral summer. From this, it is clear that B. terrestris has extensively invaded Tasmanian native vegetation. The new data provide strong evidence that B. terrestris is breeding in native vegetation in every region of Tasmania. More than 10 bumblebees were seen in one day at 153 locations in native vegetation, including 42 locations within 10 National Parks and 38 locations within the Tasmanian Wilderness World Heritage Area. Nests of B. terrestris were also found within two National Parks. These findings suggest that B. terrestris would also invade native vegetation in non-arid temperate regions of the Australian mainland, if it is introduced there.     

Colonial and geographic variations in agonistic behaviour,cuticular hydrocarbons and mtDNA of Italian populations of <Emphasis Type="Italic">Reticulitermes lucifugus</Emphasis> (Isoptera,Rhinotermitidae)

Summary The subterranean termite, Reticulitermes lucifugus (Rossi), is found throughout Italy. The purpose of this study was to assess variations among colonies collected from the north to south of Italy. A multidisciplinary approach was used including behavioral tests, cuticular hydrocarbon analysis and partial sequencing of mitochondrial DNA. Results showed that Italian R. lucifugus populations were moderately aggressive and that aggressive behavior was unrelated to intercolonial geographic distance. Analysis of cuticular hydrocarbons demonstrated no qualitative variations between colonies, but quantitative differences were found according to caste and colony. Alkene content tended to decrease from north to south. Sequencing of mtDNA indicated kinship between two Tuscan populations and R. l. corsicus. This finding is suggestive of transtyrrenian distribution of the Corsican subspecies. Moreover, distance between haplotypes appears to be associated with intercolonial geographic distance.Received 25 March 2003; revised 3 October 2003; accepted 10 November 2003.     

Distribution and flower visitation records of bumblebees in Lebanon (Hymenoptera: Apidae)

Summary

West Palearctic bumblebees are common wildflowers and crop pollinators that are well studied in their central and northern distribution ranges, but fewer information is available on their southern distribution areas. Lebanon falls on the southern limit of their distribution and no published information is available on the local bumblebees. Our study aims to produce a data baseline of the local bumblebee species. In order to do so we grouped available old records of bumblebees in Lebanon with recent author collections and produced preliminary distribution maps. We listed four species: Bombus terrestris, B. argillaceus, B. niveatus vorticosus and B. melanurus. Preliminary distribution shows that Bombus terrestris and B. argillaceus are widespread and have a large foraging range, whereas B. niveatus vorticosus and B. melanurus have a restricted distribution to altitudes above 1800 m with a smaller foraging range. The male cephalic labial gland secretions analysis of local Bombus terrestris specimens provides preliminary evidence that the local subspecies could be Bombus terrestris calabricus. Therefore, we highlight the importance of regulating foreign Bombus terrestris subspecies importation for agriculture purposes, as well as monitoring B. niveatus vorticosus and B. melanurus that are rendered vulnerable by their isolated populations.     

Induction of colony initiation by Japanese native bumble bees using cocoons of the exotic bumblebee Bombus terrestris

The colony initiation rates of Bombus hypocrita (a native Japanese bumblebee) and Bombus terrestris (a European species) foundresses were compared after 4 weeks of exposure to B. terrestris cocoons. The B. terrestris cocoons, when replaced weekly, were effective for inducing oviposition by foundresses of both species. There were no significant differences in the colony initiation rates of B. terrestris and B. hypocrita, either with the control treatment or with the cocoons. The cocoon method was also tested for five species and two subspecies of native Japanese bumblebees. The colony initiation rate was higher for foundresses of the subgenus Bombus s. str. than for foundresses of the subgenera Pyrobombus, Diversobombus, and Thoracobombus. When replaced weekly, the cocoons of B. terrestris are effective inducers of colony foundation in three Japanese native species, namely B. ignitus, B. hypocrita hypocrita, and B. h. sapporoensis.     

Genetic differentiation of continental and island populations of Bombus terrestris (Hymenoptera: Apidae) in Europe

Ten microsatellite loci and a partial sequence of the COII mitochondrial gene were used to investigate genetic differentiation in B. terrestris, a bumble bee of interest for its high-value crop pollination. The analysis included eight populations from the European continent, five from Mediterranean islands (six subspecies altogether) and one from Tenerife (initially described as a colour form of B. terrestris but recently considered as a separate species, B. canariensis). Eight of the 10 microsatellite loci displayed high levels of polymorphism in most populations. In B. terrestris populations, the total number of alleles detected per polymorphic locus ranged from 3 to 16, with observed allelic diversity from 3.8 ± 0.5 to 6.5 ± 1.4 and average calculated heterozygosities from 0.41 ± 0.09 to 0.65 ± 0.07. B. canariensis showed a significantly lower average calculated heterozygosity (0.12 ± 0.08) and observed allelic diversity (1.5 ± 0.04) as compared to both continental and island populations of B. terrestris. No significant differentiation was found among populations of B. terrestris from the European continent. In contrast, island populations were all significantly and most of them strongly differentiated from continental populations. B. terrestris mitochondrial DNA is characterized by a low nucleotide diversity: 0.18%± 0.07%, 0.20%± 0.04% and 0.27%± 0.04% for the continental populations, the island populations and all populations together, respectively. The only haplotype found in the Tenerife population differs by a single nucleotide substitution from the most common continental haplotype of B. terrestris. This situation, identical to that of Tyrrhenian islands populations and quite different from that of B. lucorum (15 substitutions between terrestris and lucorum mtDNA) casts doubts on the species status of B. canariensis. The large genetic distance between the Tenerife and B. terrestris populations estimated from microsatellite data result, most probably, from a severe bottleneck in the Canary island population. Microsatellite and mitochondrial DNA data call for the protection of the island populations of B. terrestris against importation of bumble bees of foreign origin which are used as crop pollinators.     

An overview of the Bombus terrestris (L. 1758) subspecies (Hymenoptera: Apidae)

Bombus terrestris is one of the most abundant bumblebee species in the West-Palaearctic. Its widespread domestication results in the movement of many colonies. The aim of this paper is to describe the 9 subspecies currently recognised and to list some of their most obvious characteristics. Bombus terrestris is not declining anywhere, on the contrary, its synanthropic spread through domestication may be expected. However, its autumn and winter populations rely on a restricted choice of flowers so they may be threatened locally by scrub clearance and the destruction of their favourite autumnal flowers.     

The relationships between cuticular hydrocarbon composition, faunal assemblages, inter-island distance, and population genetic variation in Tuscan Archipelago wasps

Until recently, studies examining the geographical distribution of insects in the Tuscan Archipelago have focused on paleogeography as the primary influence on species distributions. However, for flying insects such as Hymenoptera that may be able to disperse over water, current geographical location is likely to be more important in determining present distributions within the Archipelago. Here we compare mainland and island wasp populations using genetic variation and cuticular hydrocarbon composition of the vespid wasp Polistes dominulus, and species composition of wasps in the family Pompilidae. Both chemical and genetic data result in similar clustering of P. dominulus populations that reflect present geographical location. Moreover, we found current geographical distance to be significantly correlated with P. dominulus population genetic differentiation and Pompilidae faunal composition. These data suggest that dispersal over present sea distances is more important in determining population differentiation and species distribution in the Tuscan Archipelago than paleogeography.     

Glacial Effects on Sequence Divergence of Mitochondrial COII of Polyura eudamippus (Lepidoptera: Nymphalidae) in China

Sequence divergence of mitochondrial COII was analyzed in 50 specimens belonging to five subspecies of Polyura eudamippus (Lepidoptera: Nymphalidae) collected from southern China. There were nine haplotypes in the 405 bp of partial COII sequence. Distribution of the five subspecies was primarily consistent with the spatial distribution of haplotypes. The K _st statistic showed genetic differentiation among these subspecies, except between the pair of P. e. kuangtungensis and P. e. formosana, which were separated by the Taiwan Strait. This is consistent with the 10,000-year history of the Taiwan Strait, not long enough for detectable differentiation. The present distribution pattern of COII haplotypes of P. eudamippus should be shaped by the alteration of Pleistocene glaciations, and Yunnan might be the refugium of P. eudamippus in the ice age, judging from the abundant haplotypes remaining. There were two routes for P. eudamippus in the postglacial expansion, one northward to Sichuan, Chongqing, and Hubei and another eastward to the southeastern coast of mainland China and Taiwan Island. Because the haplotype of butterflies on Hainan Island (P. e. whiteheadi) was completely different from that of mainland China, it was estimated that butterflies on Hainan Island might be from the Indo-China Peninsula rather than from mainland China.     

Historical introgression and the persistence of ghost alleles in the intermediate horseshoe bat (Rhinolophus affinis)

Phylogenetic conflicts between genetic markers can help to disentangle complex histories of phylogeography and introgression among taxa. We previously proposed that the Chinese mainland subspecies of the intermediate horseshoe bat Rhinolophus affinis himalayanus colonized Hainan Island to form the subspecies R. a. hainanus. Subsequent recolonization of the mainland formed a third taxon, R. a macrurus, and a secondary contact zone with the ancestral himalayanus. To test for historical and recurrent genetic exchange between these mainland subspecies, we sampled populations of each from two parapatric zones and undertook analyses using one mtDNA marker, three nuclear genes and 14 microsatellites. Nuclear DNA, echolocation call frequencies and morphological data all recovered two taxa; however, a mtDNA phylogeny revealed two himalayanus clades, of which one clustered with macrurus, as well as some shared or related mtDNA haplotypes in eastern populations. Isolation‐with‐migration (IM) models suggested some mtDNA gene flow from macrurus to himalayanus. However, strong population structure in himalayanus raises the possibility that macrurus captured mtDNA from a coastal population of himalayanus that has since become rare or extinct. To reconcile these two sets of results, we suggest that the IM estimates might reflect historical mtDNA gene flow among populations of himalayanus, before mtDNA was subsequently captured by macrurus. Finally, microsatellite‐based ABC analyses supported the island origin of macrurus; however, mtDNA‐based ABC analyses suggest this taxon might have evolved on the mainland. Our findings highlight the importance of understanding population history and structure for interpreting hybridization and introgression events.     

Bumblebees,humble pollinators or assiduous invaders? A population comparison of foraging performance in <Emphasis Type="Italic">Bombus terrestris</Emphasis>

Worldwide trade in non-native bumblebees remains largely unrestricted despite well-documented cases where introductions of non-native bees have gone dramatically wrong. Within Europe, indiscriminate importation of non-native populations of bumblebees (Bombus terrestris) for the pollination of glasshouse crops continues on a massive scale. However, no risk assessment has been conducted for these introductions, perhaps because B. terrestris is considered a native species, so shipping populations from one region to another has been implicitly assumed to present no risk. This view is clearly unjustified because Bombus terrestris populations differ significantly in their genetic makeup as demonstrated by strong differences in coat colour and behavioural traits. Therefore, for the first time we compare an important competitive trait, namely foraging performance, between commercially available B. terrestris populations in contrasting environments. We test whether commercially reared populations differ in their nectar foraging performance and whether this is influenced by both their source environment and the one they are introduced into. We do this by means of a reciprocal transplant experiment. Strong, consistent inter-population differences in performance occurred irrespective of test location: Canary Island bees (B. t. canariensis) were superior to Sardinian bees (B. t. sassaricus), which were generally superior to mainland European bees (B. t. terrestris). These inter-population differences in performance were largely explained by inter-population variation in forager size, with larger bees being superior foragers. However, even when body size was accounted for, “native” bees were not superior to transplanted non-native bees in all but one case. We conclude that non-native populations, especially those with large foragers, can be highly competitive foragers. This could lead to their establishment and displacement of native bees. Therefore, we recommend that unregulated movements of non-native B. terrestris populations within Europe should not be carried out without a full risk assessment.     

Genetic variation in Sri Lankan leopards

Electrophoretic variation of 50 gene-enzyme systems was typed in a population of 33 captive leopards (Panthera pardus) from the island of Sri Lanka. The captive leopard population was composed of several lineages: (1) wild-caught leopards of the island subspecies (P. p. kotiya), (2) captive-born animals of the same subspecies, (3) a melanistic lineage whose founders were obtained from Malaysia (P. p. delacouri), and (4) leopards of known mixed lineage and unknown status. Two loci, APRT and PGD, were polymorphic in all samples, whereas 48 loci were invariant. Percent polymorphism (P) and percent average heterozygosity (H) were calculated as 4% and 1.4%, respectively, for the wild-caught leopards; 4% and 1.2% for the captive-born kotiya leopards; and 4% and 2.0% for the melanistic lineage. The overall results revealed a detectable decrease in genetic variability compared with a previous study of captive leopards from mainland origins. The mainland leopards had three additional polymorphic loci, ADA, ESI, and HBB. Reexamination of the TF locus using a revised protocol resolved a new allele in the sample of mainland leopards but not in the Sri Lankan sample. With this new polymorphism, recalculated P and H values for the mainland sample are 10% and 3.1%, respectively. No significant differences in polymorphic loci were observed between the leopard subspecies examined.     

Molecular survey of morphological subspecies reveals new mitochondrial lineages in Podarcis muralis (Squamata: Lacertidae) from the Tuscan Archipelago (Italy)

Recent analyses of molecular markers have significantly revised the traditional taxonomy of Podarcis species (Squamata: Lacertidae), leading to critically reconsider the taxonomic value of several subspecies described only on morphological bases. In fact, lizards often exhibit high morphological plasticity both at the intra‐specific and the intra‐population level, especially on islands, where phenotypic divergences are mainly due to local adaptation, rather than to evolutionary differentiation. The Common wall lizard Podarcis muralis exhibits high morphological variability in biometry, pholidosis values and colour pattern. Molecular analyses have confirmed the key role played by the Italian Peninsula as a multi‐glacial refuge for P. muralis, pointing out the lack of congruence between mitochondrial lineages and the four peninsular subspecies currently recognized. Here, we analyse a portion of the protein‐encoding cytochrome b gene in the seven subspecies described for the Tuscan Archipelago (Italy), in order to test whether the mitochondrial haplotypes match the morphologically based taxonomy proposed for Common wall lizard. We also compare our haplotypes with all the others from the Italian Peninsula to investigate the presence of unique genetic lineages in insular populations. Our results do not agree completely with the subspecific division based on morphology. In particular, the phylogenetic analyses show that at least four subspecies are characterized by very similar haplotypes and fall into the same monophyletic clade, whereas the other three subspecies are closer to peninsular populations from central Italy. From these results, we conclude that at least some subspecies could be better regarded as simple eco‐phenotypes; in addition, we provide an explanation for the distinctiveness of exclusive lineages found in the archipelago, which constituted a refuge for this species during last glacial periods.     

RAPD variation in Mediterranean turtle <Emphasis Type="Italic">Testudo graeca</Emphasis> L. (Testudinidae)

The polymerase chain reaction with arbitrary primers (RAPD-PCR) was used to study intraspecific variation in Mediterranean turtle Testudo graeca, which is represented by the Dagestan (T. g. pallasi) and Nikolskii (T. g. nikolskii) subspecies in Russia. To study the phylogenetic relationships, the RAPD variation was also compared in two other T. graeca subspecies (T. g. ibera and T. g. terrestris), two closely related Testudo species (T. kleinmanni and T. marginata), and Central Asian turtle Agrionemys horsfieldii. Parameters of RAPD variation showed that the sample from different geographical regions of Dagestan was more polymorphic and heterogeneous than that from Central Asia. The two samples differed in the mean number of RAPD fragments N (48.76 vs. 40.40), number of polymorphic fragments P (78.7 vs. 32.3), and within-group similarity index APS (0.607 vs. 0.784). In T. g. pallasi, no significant difference in N, P, or APS was observed between samples from different localities of Dagestan or between groups of turtles with four- or five-clawed forelegs. A dendrogram of genetic similarity between the species and subspecies under study contained two clusters, one comprising all A. horsfieldii individuals and the other, all turtles of the genus Testudo. In the latter, T. marginata and T. kleinmanni showed higher similarity to each other than to T. graeca. The four T. graeca subspecies clustered separately from each other with a high reliability, T. g. nikolskii and T. g ibera (Turkey) being more similar to each other than to T. g. terrestris or T. g. pallasi. The possible causes of the presence of four claws on forelegs of turtles from Dagestan and the relationships among members of the genus Testudo were discussed.Translated from Genetika, Vol. 40, No. 12, 2004, pp. 1628–1636.Original Russian Text Copyright © 2004 by Semyenova, Korsunenko, Vasilyev, Pereschkolnik, Mazanaeva, Bannikova, Ryskov.     

Colonization and divergence: phylogeography and population genetics of the Atlantic coast beach mice

Barrier island taxa provide an opportunity to investigate recent evolutionary processes, such as colonization and isolation of recently diverged taxa, and provide important insights into understanding contemporary diversity and the assessment of conservation units. Using rapidly evolving genetic markers (mitochondrial DNA and microsatellites), we studied the Atlantic coast beach mouse subspecies (Peromyscus polionotus decoloratus, P. p. niveiventris, and P. p. phasma). Our data indicate that each of the extant coastal subspecies (P. p. niveiventris and P. p. phasma) is comprised of unique haplotypes indicative of their isolation, while the extinct subspecies, P. p. decoloratus, contain a single haplotype, which was shared with P. p. phasma. Moreover, all the coastal haplotypes originate from a single mainland haplotype found in central Florida, USA. The microsatellite data indicated high levels of genetic structure among our sampled populations. Additionally, these data group the populations into three distinct genetic clusters, with each of the extant coastal subspecies belonging to their own cluster and the mainland individuals forming a separate cluster. The extant Atlantic coast beach mice are on separate evolutionary trajectories, thus representative of separate taxonomic units. Therefore, the data support that two extant subspecies on the Florida Atlantic coast fit the Distinct Population Segment designation and should be managed and conserved as two separate independent units.     

The role of western Mediterranean islands in the evolutionary diversification of the spotted flycatcher Muscicapa striata,a long‐distance migratory passerine species

We investigated the evolutionary history of the spotted flycatcher Muscicapa striata, a long distance migratory passerine having a widespread range, using mitochondrial markers and nuclear introns. Our mitochondrial results reveal the existence of one insular lineage restricted to the western Mediterranean islands (Balearics, Corsica, Sardinia) and possibly to the Tyrrhenian coast of Italy that diverged from the mainland lineages around 1 Mya. Mitochondrial genetic distance between insular and mainland lineages is around 3.5%. Limited levels of shared nuclear alleles among insular and mainland populations further support the genetic distinctiveness of insular spotted flycatchers with respect to their mainland counterparts. Moreover, lack of mitochondrial haplotypes sharing between Balearic birds (M. s. balearica) and Corso‐Sardinian birds (M. s. tyrrhenica) suggest the absence of recent matrilineal gene flow between these two insular subspecies. Accordingly, we suggest that insular spotted flycatchers could be treated as one polytypic species (Muscicapa tyrrhenica) that differs from M. striata in morphology, migration, mitochondrial and nuclear DNA and comprises two subspecies (the nominate and M. t. balearica) that diverged recently phenotypically and in mitochondrial DNA and but still share the same nuclear alleles. This study provides an interesting case‐study illustrating the crucial role of western Mediterranean islands in the evolution of a passerine showing high dispersal capabilities. Our genetic results highlight the role of glacial refugia of these islands that allowed initial allopatric divergence of insular populations. We hypothesize that differences in migratory and breeding phenology may prevent any current gene flow between insular and mainland populations of the spotted flycatcher that temporarily share the same insular habitats during the spring migration.     

GEOGRAPHICAL CORRELATION OF MORPHOLOGICAL VARIATION IN SILVETIA COMPRESSA (FUCACEAE,FUCALES, PHAEOPHYCEAE)

Silvetia compressa ( J. Agardh) Serrão et al. is a common member of the upper intertidal fucoid community on the Pacific coast of America from Humboldt County, California, to Punta Baja, Baja California, Mexico. A relatively narrow range of morphological variability is exhibited by most mainland populations, regardless of latitude, but some mainland populations and all insular populations participate in a complex pattern that we have attempted to analyze. A few populations on the Monterey Peninsula in which the fronds are atypically delicate were described by Setchell & Gardner as f. gracilis, to which was assigned a population from Santa Catalina Island. After comparing populations from various parts of the range of the species, including all of the Channel Islands, we conclude that two subspecies may be recognized. In subsp. compressa, which includes f. gracilis as a growth form and occurs chiefly on the mainland, the frond is robust with long tapered receptacles. In the variant subspecies, which is chiefly insular but also occurs on the coast of northern Baja California, the typical frond has slender axes as in f. gracilis, but is more densely branched and has short ellipsoidal receptacles. Comparison of nucleotide sequences from the ITS regions of rDNA revealed an identical pattern for subsp. compressa from Baja California and central California, including populations assignable to f. gracilis. By contrast, the pattern for the variant subspecies differed by 2 bp (0.3%) from that of subsp. compressa.     

Mitochondrial haplotype diversity in the tortoise species <Emphasis Type="Italic">Testudo graeca</Emphasis> from North Africa and the Middle East

Background  

To help conservation programs of the endangered spur-thighed tortoise and to gain better insight into its systematics, genetic variation and evolution in the tortoise species Testudo graeca (Testudines: Testudinidae) was investigated by sequence analysis of a 394-nucleotide fragment of the mitochondrial 12S rRNA gene for 158 tortoise specimens belonging to the subspecies Testudo graeca graeca, Testudo graeca ibera, Testudo graeca terrestris, and a newly recognized subspecies Testudo graeca whitei. A 411-nucleotide fragment of the mitochondrial D-loop was additionally sequenced for a subset of 22 T. graeca, chosen because of their 12S gene haplotype and/or geographical origin.     

Island Populations, Human Introductions and the Limitations of Genetic Analyses: the Case of the Sardinian Red Deer (Cervus elaphus corsicanus)

The Corsican red deer (Cervus elaphus corsicanus) is endemic to the Tyrrhenian islands of Corsica and Sardinia. It has been regarded as an introduced species and has allegedly been present on the islands since the beginning of the Neolithic culture some 8,000 years ago. In this review, we present the results of relevant genetic analyses and discuss their implications for the origin of C. e. corsicanus. Different genetic studies hypothesize that the most probable ancestral populations for Sardianian red deer were alternatively, the Near East, North Africa, or mainland Italy. These respective scenarios are evaluated and it is concluded that geneticists have not yet been unable to definitively solve the problem. However, a natural colonization of the Tyrrhenian islands from mainland Italy via the Tuscan archipelago is not only in accordance with palaeontological findings but also with at least some of the genetic data.