Bruno Buatois

Geographic variation of floral scent in a highly specialized pollination mutualism

Floral scents are important signals for communication between plants and pollinators. Several studies have focused on interspecific variation of these signals, but little is known about intraspecific variation in flower scent, particularly for species with wide geographic distributions. In the highly specific mutualism between Ficus species and their pollinating wasps, chemical mediation is crucial for partner encounter. Several studies show that scents, i.e. blends of volatiles, are species-specific, but no studies address interpopulation variation of scents in fig pollination mutualisms, which often have broad geographic distributions. In this study, using absorption/desorption headspace techniques, we analyzed variation in floral scent composition among three populations of each of two widely distributed Asian Ficus species. We identified more than 100 different volatile organic compounds, predominantly terpenes. In both species, significant differences were found between scent bouquets of East Asian and Indian populations. These differences are discussed in relation to geographical barriers that could disrupt gene exchange between these two areas, thereby isolating Indian populations from those of Eastern Asia.

Volatile organic compound emissions induced by the aphid Myzus persicae differ among resistant and susceptible peach cultivars and a wild relative

Little is known on aphid-induced emissions of volatile organic compounds (VOCs) from trees and particularly on their intraspecific variability in association with resistance traits. We compared VOC emissions from five peach cultivars (Prunus persica (L.) Batsch) and a wild relative (Prunus davidiana (Carrière) Franch) that differ in their level (susceptible/resistant) and type (antixenosis, antibiosis) of resistance to the green peach aphid, Myzus persicae (Sulzer). Additionally, the kinetics of VOC induction in response to aphids was compared with that by mechanical wounding. Qualitative and overall quantitative differences among peach genotypes were found in VOC emissions that were mainly composed of methyl-salicylate, farnesenes, (E)-β-ocimene and (E)-4,8-dimethyl-1,3,7-nonatriene. Irrespective of the type of resistance, all resistant genotypes had increased VOC emissions upon aphid attack, while in susceptible genotypes emissions remained low. Emission increases were highest in the genotypes that express increased aphid resistance during second infestations, which had also the highest proportions of methyl-salicylate in their emissions. VOC induction by aphids proceeded slowly with a delay of several hours. Artificial wounding of leaves did not result in emissions of aphid-induced VOCs but caused an immediate burst of green leaf volatiles and benzaldehyde. We conclude that VOC induction in resistant peach cultivars is part of a general defence syndrome that is being avoided or suppressed by M. persicae in the susceptible genotypes. The induction likely involves an aphid-specific elicitor and (methyl)-salicylate in the subsequent signalling and regulation processes that should include gene activation due to the marked delay in the emission response. The results are compared with those of the literature and discussed in view of their ecological and environmental significance.

Evolution of a genetic polymorphism with climate change in a Mediterranean landscape

Many species show changes in distribution and phenotypic trait variation in response to climatic warming. Evidence of genetically based trait responses to climate change is, however, less common. Here, we detected evolutionary variation in the landscape-scale distribution of a genetically based chemical polymorphism in Mediterranean wild thyme (Thymus vulgaris) in association with modified extreme winter freezing events. By comparing current data on morph distribution with that observed in the early 1970s, we detected a significant increase in the proportion of morphs that are sensitive to winter freezing. This increase in frequency was observed in 17 of the 24 populations in which, since the 1970s, annual extreme winter freezing temperatures have risen above the thresholds that cause mortality of freezing-sensitive morphs. Our results provide an original example of rapid ongoing evolutionary change associated with relaxed selection (less extreme freezing events) on a local landscape scale. In species whose distribution and genetic variability are shaped by strong selection gradients, there may be little time lag associated with their ecological and evolutionary response to long-term environmental change.

A case study of modified interactions with symbionts in a hybrid mediterranean orchid.

UNLABELLED PREMISE OF THE STUDY Most studies on orchid hybrids examine separately the effects of hybridization on interactions with pollinators or with mycorrhizal fungi. Here, we simultaneously investigated both interactions in the mediterranean food-deceptive Orchis simia, O. anthropophora, and their hybrid (O. ×bergonii) and tested a possible breakdown of coevolution using a multidisciplinary approach. • METHODS We compared leaf growth, seed viability, emitted scent, and mycorrhizal fungi (species and rate of infection) among these three taxa. • KEY RESULTS We show that leaf surface is greater in adult hybrids than in the parental species, suggesting a heterosis effect for vegetative growth. We demonstrate that flowers of the two parental species emit well-differentiated bouquets of volatile organic compounds, while hybrids emit larger quantities, accumulating most compounds of the two parental species. However, hybrids fail to attract pollinators and have a 10 times lower fruit set. We determined that closely related Tulasnellales are mycorrhizal in the three taxa, suggesting that the mycorrhizal partner does not impair hybrid survival. We propose an interpretative model for O. ×bergonii compared with its parents. • CONCLUSIONS In hybrids, carbon resources normally devoted to reproduction may be reallocated to the mycorrhizal symbiosis as a result of the disruption of the pollination interaction in hybrids. Higher mycorrhizal infection may in turn enhance vegetative growth and scent emission. Such interplay between the two obligate biotic interactions yields new insights into hybridization among orchids.

Geographical variations of odour and pollinators, and test for local adaptation by reciprocal transplant of two European Arum species

1. Interactions between entomophilous plants and their pollinators are one of the major factors shaping the evolution of floral features. As species are distributed in more or less connected populations, they have evolved in a geographical mosaic of co-evolution were the outcome of the plant–pollinator interaction is likely to vary as a result of local adaptations. 2. Arum italicum and Arum maculatum are two species of Araceae which deceive their fly pollinators by mimicking the odour of their oviposition sites. Whereas A. italicum is known to be pollinated by flies belonging to different families (i.e. opportunist), A. maculatum relies on only two pollinating species of the family Psychodidae throughout its European repartition area (i.e. specialist). 3. The interannual and geographical variations of pollinators and pollinator-attractive odours were described in several populations of the two species over two consecutive years. Furthermore, local adaptation to pollinators was tested by transplanting inflorescence-bearing plants between two different sites and by recording the number and composition of the insect fauna trapped inside the inflorescences during anthesis as a measure of a fitness component. 4. Pollinators and pollinator-attractive odours of the two Arum species varied in time and space, but there was no clear odour structure between populations. When transplanted, inflorescences of both species trapped the same composition and number of insects as native inflorescences at a given site; this indicates that pollinator composition is highly dependent on the local availability of insects. 5. No pattern of local adaptation was found for these two species, but local pollination conditions were shown to strongly affect the degree of geographical variations of these interactions. The lack of a clear odour geographical structure might be due to high gene flow or to similar selective pressures exerted by pollinators, and the high interindividual odour variation may be linked to the deceptive strategy adopted by the two plant species.

Nitrous Oxide (N2O) Emissions by Termites: Does the Feeding Guild Matter?

In the tropics, termites are major players in the mineralization of organic matter leading to the production of greenhouse gases including nitrous oxide (N2O). Termites have a wide trophic diversity and their N-metabolism depends on the feeding guild. This study assessed the extent to which N2O emission levels were determined by termite feeding guild and tested the hypothesis that termite species feeding on a diet rich in N emit higher levels of N2O than those feeding on a diet low in N. An in-vitro incubation approach was used to determine the levels of N2O production in 14 termite species belonging to different feeding guilds, collected from a wide range of biomes. Fungus-growing and soil-feeding termites emit N2O. The N2O production levels varied considerably, ranging from 13.14 to 117.62 ng N2O-N d-1 (g dry wt.)-1 for soil-feeding species, with Cubitermes spp. having the highest production levels, and from 39.61 to 65.61 ng N2O-N d-1 (g dry wt.)-1 for fungus-growing species. Wood-feeding termites were net N2O consumers rather than N2O producers with a consumption ranging from 16.09 to 45.22 ng N2O-N d-1 (g dry wt.)-1. Incubating live termites together with their mound increased the levels of N2O production by between 6 and 13 fold for soil-feeders, with the highest increase in Capritermes capricornis, and between 14 and 34 fold for fungus-growers, with the highest increase in Macrotermes muelleri. Ammonia-oxidizing (amoA-AOB and amoA-AOA) and denitrifying (nirK, nirS, nosZ) gene markers were detected in the guts of all termite species studied. No correlation was found between the abundance of these marker genes and the levels of N2O production from different feeding guilds. Overall, these results support the hypothesis that N2O production rates were higher in termites feeding on substrates with higher N content, such as soil and fungi, compared to those feeding on N-poor wood.

The Evolution of Communication in Two Ant-Plant Mutualisms

Myrmecophytes are plants that provide nesting sites and food to ants that protect them against herbivores. Plant signals function to synchronize ant patrolling with the probability of herbivory. We compared the communication signals in two symbioses involving ant and plant pairs that are closely related. The two plants emitted the same volatile compounds upon damage. These compounds are simple molecules common in the plant kingdom. Electroantennography revealed that the two symbiotic ants, as well as several other ant species, were able to perceive these compounds. However, workers of one species responded only to hexanal, while those of the other species responded mostly to methyl salicylate. The two signals involved in the focal symbioses are ‘cheap’ (low metabolic cost), which is consistent with theoretical predictions for the evolution of signalling between partners with convergent interests. They are also not specific, which is expected between plants and broad-spectrum predators such as ants. The fact that different signals are used in the two sister symbioses suggests different mechanisms underlying similar adaptations in the evolution of communication.

Integrative taxonomy of the fly orchid group: insights from chemical ecology

Several authors have recently stressed the need to develop an integrative approach in taxonomy, but studies applying such an approach to Mediterranean orchids are scarce. In sexually deceptive orchids from the taxonomically difficult genus Ophrys, pollination is specific and performed by male insects attracted to the flowers by sex pheromone-mimicking floral scents. Floral compounds are therefore of primary importance for reproductive isolation and species delimitations in this genus. In the fly orchid group, molecular, morphological, and ecological characters have been extensively studied, but a comprehensive survey of floral scents is still lacking. In the present study, the blends of floral compounds of its three members, Ophrys insectifera, Ophrys aymoninii, and Ophrys subinsectifera, were extracted and analyzed by gas chromatography-mass spectrometry. A total of 107 compounds were found, with a majority of saturated and unsaturated hydrocarbons. Significant differentiation, both qualitative and quantitative, was found among the three taxa. This result, pooled with those from the literature, forms a comprehensive and congruent dataset that allows us to elucidate the taxonomic rank of the three members of the fly orchid group.

Pollination along an elevational gradient mediated both by floral scent and pollinator compatibility in the fig and fig‐wasp mutualism

1Faculty of Science, University of South Bohemia, Ceske Budejovice, Czech Republic; 2Biology Centre of the Czech Academy of Sciences, Institute of Entomology, Ceske Budejovice, Czech Republic; 3Centre d’Ecologie Fonctionnelle et Evolutive (CEFE), UMR 5175, CNRS – Université de Montpellier – Université Paul Valéry Montpellier 3, EPHE, IRD, Montpellier, France; 4New Guinea Binatang Research Center, Madang, Papua New Guinea; 5Okinawa Institute of Science and Technology Graduate University, Kunigami-gun, Okinawa, Japan and 6Institute on the Environment, University of Minnesota, Saint Paul, Minnesota

Peripubertal exposure to male chemosignals accelerates vaginal opening and induces male-directed odor preference in female mice

Reproductive physiology in female mouse is profoundly affected by male odor. A well-known effect of male odor is the acceleration of puberty onset in prepubertal female mice exposed to male urine. Whether peripubertal exposure to male odor also influences female sexual behavior in adulthood is poorly known. Recently, we reported that female mice exposed to male-soiled bedding showed advanced vaginal opening associated with early expression of male-directed odor preference in adulthood. The aim of the present study is to determine whether peripubertal exposure to male urinary chemosignals affects both occurrence of vaginal opening and attraction to male odor at older age in female mice. Therefore, we exposed female mice to (1R, 5S, 7R)-3,4-dehydro-exo-brevicomin (DHB), 6-hydroxy-6-methyl-3-heptanone (HMH) and (S)-2-sec-butyl-4,5-dihydrothiazole (SBT), individually or in mixture, from postnatal day (PD) 21 to PD38 and monitored the occurrence of vaginal opening. We measured then the time that the female mice spent sniffing male and female mouse urinary volatiles at PD45. As expected, peripubertal exposure to DHB, HMH or SBT accelerated vaginal opening in female mice. In addition, we showed that exposure to a mixture of these three compounds induced expression of male-directed odor preference at PD45, contrary to the single exposure to each of these molecules. In conclusion, the volatile compounds DHB, HMH and SBT in urine of male mice influence both occurrence of vaginal opening and adult expression of male-directed odor preference in female mice.