Importance of olfactory and visuel cues of Echium for host-plant finding of the oligolectic bee Osmia adunca (Megachilidae)

Solitary bees are important pollinators of flowers. Besides nectar they collect pollen at flowers mainly to provide their larvae with food. Many bee species collect pollen only on a few closely related plant species (oligolecty) (Müller & al. 1997). Little is known about the visual and olfactorial signals they use for host-plant finding (Wcislo & Cane 1996). However, bees can olfactorily distinguish between different pollen species (von Frisch 1923), and a species-specific chemistry of pollen odour is known for some plant species (Bergström & al. 1995, Dobson & al. 1999). Further, it was shown that naïve oligolectic bees recognize their host-plant on the basis pollen volatiles (Dobson & Bergström 2000) and that flower-experienced bees could use pollen odours to assess pollen availability (Dobson & al. 1999). Besides scent, also visual cues are of relevance for host-plant finding, and bees orientate especially spectral contrasts. Biotests with dummy flowers revealed that colour contrast and not intensity and dominant wavelengths are influencing innate behavioural responses (Lunau 1990). Further it was shown that naïve bumblebees were most motivated to land on a flower when visual stimuli from the antheres are combined with olfactorial stimuli from the pollen (1992). We choose Osmia adunca P., which is highly specialized on Echium L., as a model to investigate the importance of floral cues for an oligolectic bee. Because bees learn to associate odours with reward more rapidly than visual cues (Menzel 1985), we hypothesize that scent plays a major role in attraction flower-experienced O. adunca females. We used gas chromatography to compare the scent of three Echium species with the scent of a closely related Anchusa species, and a spectrometer to compare the colour of the three Echium species. Additionally we conducted a biotest to determine the importance of visual and olfactorial signals of Echium for host-plant finding of experienced O. adunca females.Bienen besuchen Blüten, um u. a. Pollen zu sammeln, der hauptsächlich der Versorgung der Larven dient. Einige Bienenarten sammeln Pollen von nur einer oder wenigen Pflanzengattungen und werden als Oligolekten bezeichnet. Der Anlockung von Bienen dienen olfaktorische und visuelle Blütensignale. Wir postulierten, dass die eng auf Echium (Boraginaceae) spezialisierte Mauerbiene Osmia adunca (Hymenoptera: Megachilidae) gattungsspezifische visuelle und olfaktorische Blütensignale benutzt, um ihre Wirtspflanzen zu finden erkennen. Mittels chemischer Analysen (GC) konnten wir einen Echium-spezifischen Duft nachweisen, der sich deutlich von einer nah verwandten Anchusa Art unterscheidet. Biotests in einem Flugkäfig mit blütenerfahrenen, nestbauenden Weibchen ergaben weiterhin, dass Echium-Blütenduft bei der Erkennung der Pflanzen im Nahbereich eine große Rolle spielt, während visuelle Signale wahrscheinlich bei der Fernanlockung von Bedeutung sind. Die einem Reflexionsspektrometer analysierten visuellen Signale dreier Echium Arten ähnelten sehr und könnten daher vermutlich eine Funktion zur Anlockung von O. adunca haben. Weitere Tests müssen nun zeigen, welche Blütensignale naive Tiere für die Wirtspflanzenerkennung nutzen

Pollinator-attracting semiochemicals of the wasp-flower Epipactis helleborine

The orchid genus Epipactis is represented by 25 species in Europe (Richards 1982). Epipactis helleborine (L.) Crantz is the most common and widely distributed species of the genus (Wiefelspütz 1970), and is a prime example for wasp-flowers, because it is mainly pollinated by social wasps (Hymenoptera: Vespidae), like Vespula vulgaris and V. germanica (Müller 1873). Darwin (1888) already noticed that E. helleborine is almost exclusively ignored by bees and bumblebees, an observation that was confirmed in recent investigations (Keppert 2001). The flowers of E. helleborine show morphological, physiological and phenological adaptations to the visit and the pollination by Vespidae (Keppert 2001). They possess a reddish-brown or dirty purple coloration of the inflorescence (Keppert 2001), have relatively small, mostly bulbous blossoms with a broad entrance and bulbous widened, nectar-rich juice holders (Müller 1873, 1881; Schremmer 1962). Although there is much reported about wasp-pollinated flowers there is little known about the signals that are responsible for the attraction of wasps. Wiefelspütz (1970) proclaimed the statement that only the visual stimulus is responsible for the wasp attraction. Recently studies, however, assumed that odour is involved in the wasp attraction (Keppert 2001). Hölzler (2003) showed that the main attraction of the wasp-flower Epipactis for pollinators is its olfactory stimulus. It remains an unanswered question why E. helleborine flowers almost exclusively attracts social wasps, as opposed to bees and bumblebees. In this study we analysed the role of floral volatiles which are responsible for the specific attraction of social wasps. We supposed a mimicry-system in E. helleborine for the specific attraction of pollinators for the following reasons. So-called “green leaf volatiles” (GLVs) are emitted by plants while herbivorous insects, for example caterpillars, feed on them. GLVs thereby attract predators or parasitoids of the herbivorous insects (Dicke & Sabelis 1988; Turlings & al. 1990, 1995; Dicke & Vet 1999). Among the GLVs so far identified in former studies there are aldehydes, compounds that were also found in flower extracts of E. helleborine (Hölzler 2003). Therefore, we postulated that E. helleborine flowers produce GLVs in order to attract prey hunting social wasps for pollination. We performed bioassays and analysed flower odour gained to headspace-sampling using gas chromatography (GC), mass spectrometry (GC-MS) and gas chromatography coupled with electrophysiological analysis (GC-EAD) to investigate the hypothesis that E. helleborine flowers mimic “green leaf volatiles” (GLVs) to attract their pollinators.Die Orchideenart Epipactis helleborine gilt als typische Wespenblume. Die Blüten weisen Anpassungen an den Besuch und die Bestäubung durch soziale Faltenwespen (Hymenoptera: Vespidae) auf und werden häufig durch Vespula vulgaris und V. germanica bestäubt. In früheren Untersuchungen konnte gezeigt werden, dass olfaktorische Reize bei der Bestäuberanlockung eine übergeordnete Bedeutung vor optischen Reizen haben (Hölzler 2003). Die Frage, warum E. helleborine fast ausschließlich ihren optimalen Bestäuber, die soziale Faltenwespe, zur Bestäubung anlockt, und nicht auch auf andere Blütenbesucher attraktiv wirkt, ist noch unbeantwortet. Wir untersuchten die Hypothese, dass E. helleborine Blüten GLVs, die von Herbivoren befallenen Pflanzen abgegeben werden, nachahmen, um Beute jagende Wespen zur Bestäubung anzulocken. Dazu sammelten und analysierten wir Duftstoffe von Epipactis Blüten und mit Pieris-Raupen befallenen Kohl und identifizierten vier gemeinsam vorkommende GLVs. In Y-Rohrtests konnte die wespenanlockende Wirkung dieser Verbindungen nachgewiesen werden

The role of pollinator attracting scent in the sexually deceptive orchids Ophrys chestermanii, O. normanii and O. tenthredinifera

Sexual deception of male bees is one of the most remarkable mechanisms of pollination (Ackermann 1986, Proctor & al. 1996). Flowers of the orchid genus Ophrys mimic females of their pollinator species, usually bees and wasps, to attract males, which try to copulate with the flowers. During this so-called “pseudocopulation” the male removes the pollinia and transfers them to another flower to ensure pollination. Apart from visual and tactile cues, floral scent was shown to be most important for eliciting mating behaviour in males (Kullenberg 1961, Schiestl & al. 1999, Ayasse & al. 2003). Pollination in Ophrys is highly specific and usually each Ophrys species attracts only one pollinator species (Paulus & Gack 1990). The high degree of specialization provides the means of reproductive isolation between the intercrossable Ophrys-species (Ehrendorfer 1980). The complex odour-bouquets released by the flowers are species-specific and often consist of more than 100 different chemical compounds (Borg-Karlson & al. 1985, Ayasse 2006). Speciation in Ophrys-orchids may be brought about by changes in the pollinator attracting floral scent. The attraction of a new pollinator may act as a pre-zygotic isolation barrier (Stebbins 1970, Paulus & Gack 1990, Soliva & al. 2001). We investigated three sympatrically occuring Ophrys-species on Sardinia. O. chestermanii and O. normanii are endemic and are both pollinated by males of the bumblebee B. vestalis. O. tenthredinifera is pollinated by Eucera nigrilabris. There are different opinions concerning the taxonomic status of O. normanii. It has been described as an actual hybrid between O. chestermanii and O. tenthredinifera (Wood 1983). Paulus & Gack (1995) suggested that it is an own species, that either has developed from a hybrid between O. chestermanii and O. normanii or that has evolved by radiation from O. tenthredinifera. By conducting behavioural-tests with B. vestalis males, performing gas chromatographic analyses and electrophysiological studies we wanted to identify pollinator attracting scent and to clarify the taxonomic status of O. normanii.Sexualtäuschorchideen der Gattung Ophrys (Orchidaceae) imitieren die Weibchen ihrer Bestäuber in Duft, Form und Farbe. Insektenmännchen versuchen mit dem Labellum der Blüte zu kopulieren und transportieren den Pollen von Blüte zu Blüte, wodurch die Orchidee bestäubt wird. In dieser Arbeit untersuchten wir die Bestäuber anlockenden Duftstoffe der beiden endemisch auf Sardinien vorkommenden Arten O. normanii und O. chestermanii, die beide von Bombus vestalis Männchen (Hymenoptera: Apidae) bestäubt werden und von O. tenthredinifera, die Eucera nigrilabris (Hymenoptera: Apidae) zur Bestäubung anlockt. O. normanii wurde von Wood (1983) als Primärhybride beschrieben. Nach Paulus und Gack (1995) handelt es sich um eine hybridogene Art oder um eine Art die durch Abspaltung von O. tenthredinifera entstanden ist. Das Ziel der Untersuchungen war die Identifizierung Männchen-anlockender Verbindungen. Die Attraktivität der drei Arten für B. vestalis Männchen sollte Hinweise auf den Artstatus von O. normanii geben. In Biotests mit B. vestalis-Männchen lösten Blütenextrakte von O. normanii und O. chestermanii ebenso wie B. vestalis-Weibchen Kopulationsverhalten der Männchen aus, nicht jedoch Extrakte von O. tenthredinifera. Folglich handelt es sich bei O. normanii nicht um einen aktuellen Hybriden zwischen O. chestermanii und O. tenthredinifera. Ein Vergleich der GC-EAD-aktiven Duftbouquets mittels Diskriminanzanalyse ergab große Ähnlichkeiten zwischen O. normanii und O. chestermanii für die Substanzklassen der Ester, Alkohole und Fettsäuren, die daher vermutlich eine Schlüsselfunktion bei der Bestäuberanlockung haben

Temperature drives variation in flying insect biomass across a German malaise trap network

1. Among the many concerns for biodiversity in the Anthropocene, recent reports of flying insect loss are particularly alarming, given their importance as pollinators, pest control agents, and as a food source. Few insect monitoring programmes cover the large spatial scales required to provide more generalizable estimates of insect responses to global change drivers. 2. We ask how climate and surrounding habitat affect flying insect biomass using data from the first year of a new monitoring network at 84 locations across Germany comprising a spatial gradient of land cover types from protected to urban and crop areas. 3. Flying insect biomass increased linearly with temperature across Germany. However, the effect of temperature on flying insect biomass flipped to negative in the hot months of June and July when local temperatures most exceeded long-term averages. 4. Land cover explained little variation in insect biomass, but biomass was lowest in forests. Grasslands, pastures, and orchards harboured the highest insect biomass. The date of peak biomass was primarily driven by surrounding land cover, with grasslands especially having earlier insect biomass phenologies. 5. Standardised, large-scale monitoring provides key insights into the underlying processes of insect decline and is pivotal for the development of climate-adapted strategies to promote insect diversity. In a temperate climate region, we find that the positive effects of temperature on flying insect biomass diminish in a German summer at locations where temperatures most exceeded long-term averages. Our results highlight the importance of local adaptation in climate change-driven impacts on insect communities

Double-blind validation of alternative wild bee identification techniques: DNA metabarcoding and in vivo determination in the field

Over the past few decades, several investigations around the globe have reported alarming declines in the abundance and diversity of bee species. The success of effective conservation strategies targeting these important pollinators relies heavily on accurate biodiversity assessments. The shortage of taxonomic experts and the escalation of the ongoing biodiversity crisis call for the development of alternative identification tools to implement efficient monitoring programs. The validation of such techniques is crucial to ensure that they provide results comparable to those of traditional morphotaxonomy. Here we performed two double-blind experiments to evaluate the accuracy of a pair of new techniques used for wild bee identification: DNA metabarcoding and in vivo identification in the field. The methods were tested on sets of wild bees from Germany and their results compared against evaluations done by panels of bee experts using traditional morphotaxonomy. On average the congruency of species identification between metabarcoding and morphotaxonomy was 88.98% across samples (N = 10), while in vivo identification and morphotaxonomy were 91.81% congruent (N = 7) for bees considered feasible for in vivo identification in the field. Traditional morphotaxonomy showed similar congruencies when compared to itself: 93.65% in the metabarcoding study and 92.96% in the in vivo study. Overall, these results support both new methods as viable alternatives to traditional microscopy-based assessment, with neither method being error-free. Metabarcoding provides a suitable option to analyze large numbers of specimens in the absence of highly trained taxonomic experts, while in vivo identification is recommended for repeated long-term monitoring, and when working in areas where the sampling of individuals could threaten local populations of endangered wild bee species. Further research is still needed to explore the potential of both techniques for conservation management and wildlife monitoring, as well as to overcome their current limitations as taxonomic tools

Signal and reward in wild fleshy fruits : does fruit scent predict nutrient content?

The study examines the relationship between olfactory signals and nutrient rewards in 28 fruiting plant species in Madagascar. Previous work has shown that lemurs are the main seed dispersers in the ecosystem, relying on fruit scent to identify ripe fruits. The relative amounts of four chemical classes in fruit scent are measured using gas chromatography and mass spectrometry, as well as the relative amounts of sugar and protein in fruit pulp. The project tests the phylogenetic signal to examine whether closely related taxa tend to be similar, and compares the nutritional content of lemur‐ and bird‐dispersed fruits. The relationships reported here are across species, not within them

Where are we now with European forest multi-taxon biodiversity and where can we head to?

The European biodiversity and forest strategies rely on forest sustainable management (SFM) to conserve forest biodiversity. However, current sustainability assessments hardly account for direct biodiversity indicators. We focused on forest multi-taxon biodiversity to: i) gather and map the existing information; ii) identify knowledge and research gaps; iii) discuss its research potential. We established a research network to fit data on species, standing trees, lying deadwood and sampling unit description from 34 local datasets across 3591 sampling units. A total of 8724 species were represented, with the share of common and rare species varying across taxonomic classes: some included many species with several rare ones (e.g., Insecta); others (e.g., Bryopsida) were represented by few common species. Tree-related structural attributes were sampled in a subset of sampling units (2889; 2356; 2309 and 1388 respectively for diameter, height, deadwood and microhabitats). Overall, multitaxon studies are biased towards mature forests and may underrepresent the species related to other developmental phases. European forest compositional categories were all represented, but beech forests were overrepresented as compared to thermophilous and boreal forests. Most sampling units (94%) were referred to a habitat type of conservation concern. Existing information may support European conservation and SFM strategies in: (i) methodological harmonization and coordinated monitoring; (ii) definition and testing of SFM indicators and thresholds; (iii) data-driven assessment of the effects of environmental and management drivers on multi-taxon forest biological and functional diversity, (iv) multi-scale forest monitoring integrating in-situ and remotely sensed information. Forest biodiversity Multi-taxon Sustainable management Biodiversity conservation Forest stand structurepublishedVersio