Bombus Dahlbomii Research Articles

Chromosome-Level Assembly and Annotation of the Genome of the Endangered Giant Patagonian Bumble Bee Bombus dahlbomii.

This article describes a genome assembly and annotation for Bombus dahlbomii, the giant Patagonian bumble bee. DNA from a single, haploid male collected in Argentina was used for PacBio (HiFi) sequencing, and Hi-C technology was then used to map chromatin contacts. Using Juicer and manual curation, the genome was scaffolded into 18 main pseudomolecules, representing a high-quality, near chromosome-level assembly. The sequenced genome size is estimated at 265 Mb. The genome was annotated based on RNA sequencing data of another male from Argentina, and BRAKER3 produced 15,767 annotated genes. The genome and annotation show high completeness, with >95% BUSCO scores for both the genome and annotated genes (based on conserved genes from Hymenoptera). This genome provides a valuable resource for studying the biology of this iconic and endangered species, as well as for understanding the impacts of its decline and designing strategies for its preservation.

Occurrence of bee viruses and pathogens associated with emerging infectious diseases in native and non-native bumble bees in southern Chile

The invasion of non-native bees to new ecological territories could spread novel pathogens causing emerging infectious diseases (EIDs) in native species. We provide novel information on the prevalence, load, and co-infection network of honey bee viruses, trypanosoma, microsporidia and neogregarinorida pathogens in native Bombus dahlbomii and non-native Bombus terrestris and Bombus ruderatus. Apicystis bombi and Crithidia bombi were highly prevalent (> 78%) in three bumble bee species, with high loads of these pathogens. Nosema bombi was detected only in B. terrestris (37%) and B. ruderatus (15%). Lotmaria passim was detected in low prevalence (< 6%) and low loads in three bumble bee species. Deformed wing virus (genotype A) was detected only in B. terrestris (20%) and B. ruderatus (6%). Black queen cell virus was detected in B. terrestris (34%), B. ruderatus (22%) and B. dahlbomii (23%). Chronic bee paralysis virus, Kashmir bee virus and Acute bee paralysis virus were detected with low prevalence (7%) and titers in the three bumble bee species. The proximity of apiaries and collection sites was not a significant factor in the presence of viruses in bumble bees. The three bumble species were found to be co-infected with Apicystis bombi and C. bombi; a significant positive correlation was found between these two parasites, especially in B. terrestris. Multiple infections with N. bombi, A. bombi, C. bombi and viruses in B. terrestris and B. ruderatus were also detected. This suggests that the invasion and successful establishment of exotic bumble bees in a new area also entails the possible establishment of the pathogens that they carry, which could also be present in native bee species. This finding evidences a potential link between the population decline of B. dahlbomii and the pathogens that were detected with high levels and prevalence.

Noticia inquietante: ausencia del abejorro Bombus (Fervidobombus) dahlbomii guérin-méneville (Hymenoptera: Apidae) en la primavera 2017-verano 2018 de la ciudad de Punta Arenas (53 o 10&amp;#8217;S; 70 o 55&amp;#8217;O), Magallanes, Chile meridional, con antecedentes históricos de la especie

Se informa sobre la ausencia del abejorro Bombus dahlbomii Guerin-Meneville en Punta Arenas (Region de Magallanes, Chile) durante la primavera y verano (2017-2018). El analisis del estado actual de la flora y vegetacion, en adicion con el registro climatico y teoria de la ventana temporal no son elementos suficientes para explicar la ausencia de este apido. Se proporcionan los antecedentes historicos para completar su ficha nomenclatural y establecer un posible estatus de no continuidad de su existencia especifica. Se hipotetiza que este abejorro, por razones desconocidas, estaria iniciando su periodo de extincion

Molecular detection of the tracheal mite Locustacarus buchneri in native and non-native bumble bees in Chile

Bumble bees (Bombus spp.) are considered one of the most important pollinators of native, non-native, and domesticated plants. Pathogens and parasites, such as the tracheal mite Locustacarus buchneri (Podapolipidae), are harmful to bumble bees. We developed primers based on the mitochondrial CO1 gene for the specific detection of L. buchneri directly from samples of native Bombus dahlbomii and non-native B. terrestris and B. ruderatus. PCR products amplified and sequenced for these primers shared 100 % identity among themselves and around 99–100 % identity with other sequences of L. buchneri reported in Belgium, the Netherlands, and Japan. Bombus terrestris was the most infected species (41 %), followed by B. ruderatus (31 %) and B. dahlbomii (23 %). This is the first report involving L. buchneri in native and non-native bumble bees in Chile. Our described PCR-based detection offers a feasible, specific, and rapid method for the screening of L. buchneri in any bumble bee species.

Comparative flight morphology in queens of invasive and native Patagonian bumblebees (Hymenoptera: Bombus)

Since its introduction in Chile, the European Bombus terrestris L. (Hymenoptera: Apidae) has progressively reduced the abundance of the native Patagonian bumblebee, Bombus dahlbomii Guérin. Because an important cause of successful invasion of a species may depend on a potentially advantageous phenotype, we studied morphologies related to flight performance (flight muscle ratio (FMR), wing loading (WL), excess power index (EPI, which integrates FMR and WL) and wing aspect ratio (AR)) in the queens of the two species. Previous empirical studies showed that greater FMR, AR and EPI, and lower WL increase flight performance. In the Patagonian Chilean fjord where the study was carried out, B. dahlbomii was 40% heavier than B. terrestris, a difference theoretically allowing the queens of the native species to take off with heavier loads, despite the fact that the two species have virtually identical FMRs. However, FMR negatively depended on body mass at the intra-specific level. The total wing area was 35% greater in B. dahlbomii, but the difference in forewing length was only of 16%. Once taken into account the effect of body size, WL, was significantly lower in B. terrestris. AR increased with body mass and did not differ between species. EPI was weakly but significantly higher in B. terrestris. Experiments formally linking such parameters with flight performance may help to explain the observed quick and wide spread of this alien species in Patagonia in the last few years.

Decadal trends in the pollinator assemblage of Eucryphia cordifolia in Chilean rainforests.

Long-term studies of plant-pollinator interactions are almost nonexistent in the scientific literature. The objective of the present study was to determine changes and trends in the pollinator assemblage of ulmo (Eucryphia cordifolia; Cunoniaceae), a canopy-emergent tree found in Chilean temperate rainforests. We assessed the temporal variability of the pollinator assemblage and identified possible modulators of the observed temporal shifts. We sampled insect visitors to the flowers of 16 individual trees of E. cordifolia during 10 consecutive flowering seasons (2000-2009), recording a total of 137 pollinator species with a mean number of species per year of 44. Only three pollinator species (2.2%) were recorded every year. Two bee species accounted for 50% of all insect visits to flowers. One bee species, Bombus dahlbomii (native), was dominant in one season, whereas Apis mellifera (exotic) dominated during the next season. These interannual shifts in population abundances presented first-order dynamics that were characterized by oscillations with a period of 2 years. Changes in the abundances of the dominant pollinators, as well as differences in temperature and precipitation during insect emergence and flowering, led to a nested temporal structure of pollinator composition. Furthermore, the abundances of less common pollinators were sensitive to the abundance of the dominant bee species and to monthly maximum temperatures and the average precipitation during spring and summer. Based on our results and those from other studies, we predict a decline in the numbers of Bombus dahlbomii and nondominant native pollinators in response to new exotic arrivals.

Invasive bees promote high reproductive success in Andean orchids

Most non-autogamous orchids rely on either a single, or on a small number of pollinators to set fruit. The nectar-rewarding orchid Brachystele unilateralis and the nectarless and deceit-pollinated Chloraea virescens are restricted to the Southern Andes, where a single native and endemic bumblebee (Bombus dahlbomii) occured until the introduction of honeybees and highly invasive European bumblebees. Here, we describe the floral features, breeding system, pollinator activity and fruiting success of these orchid species. Both are self-compatible but pollinator-dependent, i.e., unable to set fruit and seed in the absence of pollinators. Field observations revealed that most of their current reproductive success is due to the introduced Bombus terrestris, Bombus ruderatus and Apis mellifera bees. The only native pollinator recorded was B. dahlbomii, but this bee was rarely observed due to its steady decline since the introduction of the alien Bombus species. The observed natural fruiting success per inflorescence in both studied species proved to be remarkably high by orchid standards: 83% and 66%, respectively. These results suggest that sexual reproduction in these orchids is not necessarily threatened by the decline of their native pollinator since pollination is successfully achieved by introduced bees. This provides a plausible better outlook for these and other regional, bumblebee-pollinated orchids and raises caution on eventual conservation policies involving the management of these introduced bee species. To our knowledge, this is the first report of native orchid pollinators being effectively replaced by invasive bumblebees and, furthermore, the first description of the pollination biology of a species of Brachystele.

Detrimental effects of volcanic ash deposition on bee fauna and plant-pollinator interactions

Volcanic eruptions are large-scale natural disturbances, which can negatively affect insect fauna and the ecological interactions in which they are involved. The 2011 eruption of the volcanic complex Puyehue Cordon-Caulle (PCC) produced the deposition of 950 million tons of ash on Argentine Patagonia, creating an ash layer of varying thickness. Although experimental studies confirmed that PCC volcanic ash negatively affects survival and behavior in many insect taxa, including bees, the effects of ash deposition on the plant-pollinator interactions (PPI) of this group of insects in natural landscapes remained untested. We evaluated the effect of the gradient of increasing ash layer thickness (0-15cm) on: (1) number of wild bees visiting flowers and total bee richness in 16 raspberry fields after the eruption, (2) number of native ( Bombus dahlbomii) and invasive (B. terrestris and B. ruderatus) bumble bees foraging on wild flowers in 10 sites before and after the eruption, and (3) the proportion of flowers (i.e. papilionaceous flowers visited for first time by large bees) in 32 populations of the invasive shrub scotch broom (Cytisus scoparius), before and after the eruption. With the increase of ash deposition, we found a consistent and significant decrease in (1) the number of wild bees and total bee richness visiting raspberry flowers; (2) the number of bumble bees, particularly B. terrestris, visiting wild flowers; and (3) the proportion of triggered flowers of scotch broom. Thus, volcanic eruptions can exert a detrimental effect on bee fauna and concomitant PPI, with a potential cascade effect on the pollination service to crops, the spread of invasive bumble bees, and the pollination success of invasive plants. (Keywords: Bombus terrestris, bumble bees, large scale disturbance, pollination, Puyehue Cordon-Caulle Volcano, raspberry, scotch broom)

Detrimental effects of volcanic ash deposition on bee fauna and plant-pollinator interactions

Ecología Austral, 24:42-50 (2014)Volcanic eruptions are large-scale natural disturbances, which can negatively affect insect fauna and the ecological interactions in which they are involved. The 2011 eruption of the volcanic complex Puyehue Cordón-Caulle (PCC) produced the deposition of 950 million tons of ash on Argentine Patagonia, creating an ash layer of varying thickness. Although experimental studies confirmed that PCC volcanic ash negatively affects survival and behavior in many insect taxa, including bees, the effects of ash deposition on the plant-pollinator interactions (PPI) of this group of insects in natural landscapes remained untested. We evaluated the effect of the gradient of increasing ash layer thickness (0-15cm) on: (1) number of wild bees visiting flowers and total bee richness in 16 raspberry fields after the eruption, (2) number of native (Bombus dahlbomii) and invasive (B. terrestris and B. ruderatus) bumble bees foraging on wild flowers in 10 sites before and after the eruption, and (3) the proportion of "triggered" flowers (i.e. papilionaceous flowers visited for first time by large bees) in 32 populations of the invasive shrub scotch broom (Cytisus scoparius), before and after the eruption. With the increase of ash deposition, we found a consistent and significant decrease in (1) the number of wild bees and total bee richness visiting raspberry flowers; (2) the number of bumble bees, particularly B. terrestris, visiting wild flowers; and (3) the proportion of triggered flowers of scotch broom. Thus, volcanic eruptions can exert a detrimental effect on bee fauna and concomitant PPI, with a potential cascade effect on the pollination service to crops, the spread of invasive bumble bees, and the pollination success of invasive plants.

The invasion of southern South America by imported bumblebees and associated parasites

The Palaearctic Bombus ruderatus (in 1982/1983) and Bombus terrestris (1998) have both been introduced into South America (Chile) for pollination purposes. We here report on the results of sampling campaigns in 2004, and 2010-2012 showing that both species have established and massively expanded their range. Bombus terrestris, in particular, has spread by some 200 km year(-1) and had reached the Atlantic coast in Argentina by the end of 2011. Both species, and especially B. terrestris, are infected by protozoan parasites that seem to spread along with the imported hosts and spillover to native species. Genetic analyses by polymorphic microsatellite loci suggest that the host population of B. terrestris is genetically diverse, as expected from a large invading founder population, and structured through isolation by distance. Genetically, the populations of the trypanosomatid parasite, Crithidia bombi, sampled in 2004 are less diverse, and distinct from the ones sampled later. Current C. bombi populations are highly heterozygous and also structured through isolation by distance correlating with the genetic distances of B. terrestris, suggesting the latter's expansion to be a main structuring factor for the parasite. Remarkably, wherever B. terrestris spreads, the native Bombus dahlbomii disappears although the reasons remain unclear. Our ecological and genetic data suggest a major invasion event that is currently unfolding in southern South America with disastrous consequences for the native bumblebee species.

Genetic variability of the neogregarine Apicystis bombi, an etiological agent of an emergent bumblebee disease.

The worldwide spread of diseases is considered a major threat to biodiversity and a possible driver of the decline of pollinator populations, particularly when novel species or strains of parasites emerge. Previous studies have suggested that populations of introduced European honeybee (Apis mellifera) and bumblebee species (Bombus terrestris and Bombus ruderatus) in Argentina share the neogregarine parasite Apicystis bombi with the native bumblebee (Bombus dahlbomii). In this study we investigated whether A. bombi is acting as an emergent parasite in the non-native populations. Specifically, we asked whether A. bombi, recently identified in Argentina, was introduced by European, non-native bees. Using ITS1 and ITS2 to assess the parasite’s intraspecific genetic variation in bees from Argentina and Europe, we found a largely unstructured parasite population, with only 15% of the genetic variation being explained by geographic location. The most abundant haplotype in Argentina (found in all 9 specimens of non-native species) was identical to the most abundant haplotype in Europe (found in 6 out of 8 specimens). Similarly, there was no evidence of structuring by host species, with this factor explaining only 17% of the genetic variation. Interestingly, parasites in native Bombus ephippiatus from Mexico were genetically distant from the Argentine and European samples, suggesting that sufficient variability does exist in the ITS region to identify continent-level genetic structure in the parasite. Thus, the data suggest that A. bombi from Argentina and Europe share a common, relatively recent origin. Although our data did not provide information on the direction of transfer, the absence of genetic structure across space and host species suggests that A. bombi may be acting as an emergent infectious disease across bee taxa and continents.

Rapid ecological replacement of a native bumble bee by invasive species

Despite rising global concerns over the potential impacts of non‐native bumble bee (Bombus spp) introductions on native species, large‐scale and long‐term assessments of the consequences of such introductions are lacking. Bombus ruderatus and Bombus terrestris were sequentially introduced into Chile and later entered Argentina's Patagonian region. A large‐scale survey in Patagonia reveals that, in 5 years post‐arrival, the highly invasive B terrestris has become the most abundant and widespread Bombus species, and its southward spread is concurrent with the geographic retraction of the only native species, Bombus dahlbomii. Furthermore, a 20‐year survey of pollinators of the endemic herb Alstroemeria aurea in northern Patagonia indicates that B ruderatus and B terrestris have replaced B dahlbomii, formerly the most abundant pollinator. Although the decline's underlying mechanisms remain unknown, the potential roles of exploitative competition and pathogen co‐introduction cannot be ruled out. Given that invasive bumble bees can rapidly extirpate native congeners, further introductions should be discouraged.

Assessing the impact of the invasive buff-tailed bumblebee (Bombus terrestris) on the pollination of the native Chilean herb Mimulus luteus

The arrival of exotic pollinators to new habitats may introduce new patterns of floral preference and foraging behavior that modify the structure of the resident plant–pollinator community. The aim of this paper is to examine the potential impact of the exotic bumblebee Bombus terrestris on the pollination service provided by the native pollinator assemblage of the herb Mimulus luteus. The study was performed in a high-elevation locality in the Chilean Andes during the summer seasons of 2010, 2011, and 2012. We recorded visitation rate, and the number of pollen grains transported on the body of B. terrestris and native pollinators and the pollen deposition on the stigmas of M. luteus. Pollinator effectiveness (pollen deposited × visitation rate) was compared among species. Results revealed that B. terrestris was an inefficient pollinator, due to the low amount of pollen delivered on stigmas and the low and intermittent visitation rate across years. The parallel inter-annual variation in the visitation rate of B. terrestris and the native bumblebee Bombus dahlbomii suggests that the integration of B. terrestris had no important consequences for the congeneric species. In general, B. terrestris accounted for a low proportion of the pollen transfer in M. luteus, reaching 4.6 % in 2010, absence of effect in 2011, and 0.01 % in 2012. These results suggest that in spite of being a quickly spreading species in Chile, B. terrestris is still in the initial phase of invasion in this area.

Hidden in taxonomy: Batesian mimicry by a syrphid fly towards a Patagonian bumblebee

Abstract Batesian mimicry has been repeatedly reported in syrphid flies (Diptera: Syrphidae), with noxious Hymenoptera identified as the models, including bumblebees (Hymenoptera: Aculeata). Despite the number of detailed studies of bumblebee mimics from the Holarctic, only minimal biological and ecological information is available for the same phenomenon in most other biogeographical regions. Here, we analyse in detail a case of Batesian mimicry by the syrphid fly Aneriophora aureorufa Philippi towards the bumblebee Bombus dahlbomii Guérin from Patagonia, a relationship only briefly noted previously in taxonomic studies. A. aureorufa possesses strikingly similar red tawny colouration to the highly hairy body of its model, and somewhat resembles it also in size. Cluster analysis suggests that the mimicry is more pronounced towards larger rather than smaller bumblebee workers. The mimicry is visually very good, but there was no evidence of a behavioural component. Foraging activity of both species seems to be largely restricted to the endemic plant Eucryphia cordifolia. The time spent on flowers was much higher in syrphid flies than in B. dahlbomii and other pollinators, and the time spent between flower visits largely overlapped between all the tested species. The endemic distribution, the apparent plant specialisation, and the invasion of alien bumblebees, make B. dahlbomii and A. aureorufa potentially threatened in some parts of the austral American forests, a priority conservation area.

Changes in wing length in the pollinator Bombus dahlbomii occurring with the fragmentation of the Maulino forest, Chile

La fragmentacion del habitat puede afectar el diseno morfologico de los insectos voladores de distintas maneras, a traves de cambios en los costos y beneficios de los patrones de dispersion. Como resultado, es probable que alas mas grandes puedan ser favorecidas cuando los recursos necesarios se encuentren dispersos a lo largo del paisaje fragmentado. En esta investigacion se puso a prueba esta prediccion en Bombus dahlbomii (Guerin), un polinizador nativo de Chile asociado al bosque Maulino fragmentado. Mediante trampas colgantes, se capturaron individuos en el bosque nativo continuo, en cuatro pequenos fragmentos y en dos plantaciones de Pinus radiata (matriz). A cada individuo colectado se le midio su tamano corporal (mm) y el largo de su primer y segundo par de alas (mm). En total, fueron capturados 280 individuos. El tamano corporal y el largo de ambos pares de alas fue mayor en los fragmentos y en la matriz, en comparacion con el bosque continuo. Ademas, se observo una relacion positiva y significativa entre la longitud de las alas y el tamano corporal en todos los habitats, sin embargo, la varianza explicada fue menor en el bosque continuo que en los fragmentos y la matriz. Estos resultados sugieren que la fragmentacion del bosque Maulino podria favorecer fenotipos con alas mas grandes, probablemente con mayores capacidades de vuelo para la busqueda de recursos escasos durante el forrajeo.

Differential effects of anthropogenic edges and gaps on the reproduction of a forest-dwelling plant: The role of plant reproductive effort and nectar robbing by bumblebees

Forest fragmentation produces sharp changes in the spatial configuration of remnant forest patches, which include the increasing influence of patch edges on the interior of forest patches (‘edge effects’). Human activities responsible for forest fragmentation tend also to change the internal characteristics of remnant patches, for example, through the creation of gaps by selective logging. While edges and gaps can be expected to cause comparable changes in the micro-environmental conditions of the forest, their effects on forest-dwelling species and their interactions are not necessarily comparable. This study compares the effect of forest edges and anthropogenic gaps on the reproductive success of a self-incompatible epiphytic plant (Mitraria coccinea), mediated by changes in its relationship with mutualists (pollinators) and antagonists (flower larcenists). Mitraria coccinea's flowers are pollinated by hummingbirds and robbed by the bumblebee Bombus dahlbomii. Edges and gaps had comparable positive effects on flower production and fruit set; however, nectar robbing was up to sevenfold higher in patches with numerous gaps and resulted in lower reproductive success (fruit set and total fruit crop). Forest fragmentation studies should therefore avoid treating forest remnants as homogeneous units (i.e. focusing exclusively on their characteristics and connectivity), without taking into account the internal heterogeneity caused by anthropogenic pressures (e.g. gaps and edge effects).

Comparative nectar-foraging behaviors and efficiencies of an alien and a native bumble bee

Resource preemption by alien organisms can contribute to their invasion success and the demise of functionally equivalent native species, particularly when opportunistic foraging by aliens results in more efficient exploitation. In forests of NW Patagonia, the only native bumble bee and major pollinator, Bombus dahlbomii, declined almost to extinction as the alien B. ruderatus increased in abundance since its first appearance about 17 years ago. To explore whether resource competition might have driven this displacement we studied the behav- ior and foraging efficiency of both bumble bees while they harvested nectar from flowers of Alstroemeria aurea, the main summer food resource in the forests of NW Patagonia. We compared the nectar content of flowers that bees selected, recently visited, and rejected with that of randomly-chosen neighboring flowers and assessed differences in visitation rates. The native bumble bee selects flowers with abundant nectar and mostly exploits nectar-rich flower patches by rejecting a higher proportion of flowers with little or no nectar. On the other hand, the alien bumble bee discriminated less with respect to sugar content per visited flower, but visited more flowers per minute. Workers of the native bumble bee harvested *70% more sugar per unit of time than those of the alien species in absolute terms, and a similar amount when sugar harvested was expressed as a percentage of body mass. In contrast to expectation, the opportu- nistic foraging of the alien bumblebee was not more efficient and therefore cannot explain the ecological extinction of the native species through exploitative competition. These findings suggest that the displace- ment of the native species by the alien may be driven by other factors, such as the associated introduction of novel diseases or parasites.

Can red flowers be conspicuous to bees?Bombus dahlbomiiand South American temperate forest flowers as a case in point

It has been argued that trichromatic bees with photoreceptor spectral sensitivity peaks in the ultraviolet (UV), blue and green areas of the spectrum are blind to long wavelengths (red to humans). South American temperate forests (SATF) contain a large number of human red-looking flowers that are reported to be visited by the bumblebee Bombus dahlbomii. In the present study, B. dahlbomii's spectral sensitivity was measured through electroretinogram (ERG) recordings. No extended sensitivity to long wavelengths was found in B. dahlbomii. The spectral reflectance curves from eight plant species with red flowers were measured. The color loci occupied by these flowers in the bee color space was evaluated using the receptor noise-limited model. Four of the plant species have pure red flowers with low levels of chromatic contrast but high levels of negative L-receptor contrast. Finally, training experiments were performed in order to assess the role of achromatic cues in the detection and discrimination of red targets by B. dahlbomii. The results of the training experiments suggest that the bumblebee relies on achromatic contrast provided by the L-receptor to detect and discriminate red targets. These findings are discussed in the context of the evolutionary background under which the relationship between SATF species and their flower visitors may have evolved.

Displacement of a native by an alien bumblebee: lower pollinator efficiency overcome by overwhelmingly higher visitation frequency

Biological invasions might constitute a major threat to mutualisms. Introduced pollinators might competitively displace their native counterparts, which in turn affects the pollination of native plants, if native and alien visitors differ in pollinator effectiveness. Since its invasion in 1994 into south-west Argentina, the introduced European bumblebee Bombus ruderatus has continuously increased in abundance, along with a simultaneous decrease in the abundance of the native Bombus dahlbomii. The latter is the only native bumblebee species of the temperate forests of southern South America, and the main pollinator of the endemic herb Alstroemeria aurea. In order to evaluate the impact of the ongoing displacement of the native by the alien bumblebee, we compared the pollinator effectiveness (i.e., the combination of pollinator efficiency per visit and visitation frequency) between both bumblebee species, as well as related pollinator traits that might account for potential differences in pollinator efficiency. Native Bombus dahlbomii, which has a larger body and spent more time per flower, was the more efficient pollinator compared to Bombus ruderatus, both in terms of quantity and quality of pollen deposited per visit. However, Bombus ruderatus was a much more frequent flower visitor than Bombus dahlbomii. As a consequence, Bombus ruderatus is nowadays a more effective pollinator of A. aurea than its native congener. Despite the lack of evidence of an increase in seed set at the population level, comparisons with historical records of Bombus dahlbomii abundances prior to Bombus ruderatus' invasion suggest that the overall pollination intensity of A. aurea might in fact have risen as a consequence of this invasion. Field experiments like these, that incorporate the natural variation in abundance of native and alien species, are powerful means to demonstrate that the consequences of invasions are more complex than previous manipulated and controlled experiments have suggested.

Is floral diversification associated with pollinator divergence? Flower shape, flower colour and pollinator preference in Chilean Mimulus.

Adaptation to different pollinators is thought to drive divergence in flower colour and morphology, and may lead to interspecific reproductive isolation. Floral diversity was tested for association with divergent pollinator preferences in a group of four closely related wildflower species: the yellow-flowered Mimulus luteus var. luteus and the red-pigmented M. l. variegatus, M. naiandinus and M. cupreus. Patterns of pollinator visitation were evaluated in natural plant populations in central Chile, including both single-species and mixed-species sites. Floral anthocyanin pigments were identified, and floral morphology and nectar variation were quantified in a common garden experiment using seeds collected from the study sites. Mimulus l. luteus, M. l. variegatus and M. naiandinus are morphologically similar and share a single generalist bumblebee pollinator, Bombus dahlbomii. Mimulus cupreus differs significantly from the first three taxa in corolla shape as well as nectar characteristics, and had far fewer pollinator visits. This system shows limited potential for pollinator-mediated restriction of gene flow as a function of flower colour, and no evidence of transition to a novel pollinator. Mimulus cupreus may experience reduced interspecific gene flow due to a lack of bumblebee visitation, but not because of its red pigmentation: rare yellow morphs are equally undervisited by pollinators. Overall, the results suggest that factors other than pollinator shifts may contribute to the maintenance of floral diversity in these Chilean Mimulus species.