Acetoin is a key odor for resource location in the giant robber crab Birgus latro.

The terrestrial and omnivorous robber crab Birgus latro inhabits islands of the Indian Ocean and the Pacific Ocean. The animals live solitarily but occasionally gather at freshly opened coconuts or fructiferous arenga palms. By analyzing volatiles of coconuts and arenga fruit, we identified five compounds, including acetoin, which are present in both food sources. In a behavioral screen performed in the crabs' habitat, a beach on Christmas Island, we found that of 15 tested fruit compounds, acetoin was the only volatile eliciting significant attraction. Hence, acetoin might play a key role in governing the crabs' aggregation behavior at both food sources.

Giant robber crabs monitored from space: GPS-based telemetric studies on Christmas Island (Indian Ocean).

We investigated the navigational capabilities of the world's largest land-living arthropod, the giant robber crab Birgus latro (Anomura, Coenobitidae); this crab reaches 4 kg in weight and can reach an age of up to 60 years. Populations are distributed over small Indo-Pacific islands of the tropics, including Christmas Island (Indian Ocean). Although this species has served as a crustacean model to explore anatomical, physiological, and ecological aspects of terrestrial adaptations, few behavioral analyses of it exist. We used a GPS-based telemetric system to analyze movements of freely roaming robber crabs, the first large-scale study of any arthropod using GPS technology to monitor behavior. Although female robber crabs are known to migrate to the coast for breeding, no such observations have been recorded for male animals. In total, we equipped 55 male robber crabs with GPS tags, successfully recording more than 1,500 crab days of activity, and followed some individual animals for as long as three months. Besides site fidelity with short-distance excursions, our data reveal long-distance movements (several kilometers) between the coast and the inland rainforest. These movements are likely related to mating, saltwater drinking and foraging. The tracking patterns indicate that crabs form route memories. Furthermore, translocation experiments show that robber crabs are capable of homing over large distances. We discuss if the search behavior induced in these experiments suggests path integration as another important navigation strategy.

Multiple gene genealogies and species recognition in the ectomycorrhizal fungus Paxillus involutus.

Paxillus involutus (basidiomycetes, Boletales) is a common ectomycorrhizal fungus in the Northern Hemisphere. The fungus displays significant variation in phenotypic characters related to morphology, physiology, and ecology. Previous studies have shown that P. involutus contains several intersterility groups and morphological species. In this study, we have used concordance of multiple gene genealogies to identify genetically isolated species of P. involutus. Fragments from five protein coding genes in 50 isolates of P. involutus collected from different hosts and environments in Europe and one location in Canada were analysed using phylogenetic methods. Concordance of the five gene genealogies showed that P. involutus comprises at least four distinct phylogenetic lineages: phylogenetic species I (with nine isolates), II (33 isolates), III (three isolates), and IV (five isolates). The branches separating the four species were long and well supported compared with the species internodes. A low level of shared polymorphisms was observed among the four lineages indicating a long time since the genetic isolation began. Three of the phylospecies corresponded to earlier identified morphological species: I to P. obscurosporus, II to P. involutus s. str., and III to P. validus. The phylogenetic species had an overlapping geographical distribution. Species I and II differed partly in habitat and host preferences.

Ectomycorrhizal mycelial species composition in apatite amended and non-amended mesh bags buried in a phosphorus-poor spruce forest.

We studied the effect of apatite amendment on ectomycorrhizal (EM) mycelial biomass production and species composition in a phosphorus-poor spruce forest using sand-filled mesh bags. Control and apatite-amended bags were buried in pairs in the lower part of the organic horizon for one growth season. DNA extraction, PCR of the ITS region, cloning and random sequencing were used to examine the fungal species composition in each bag. Sequences were identified by comparison with the UNITE database and GenBank. Our study confirmed previous results that the major fungal ingrowth in mesh bags was of EM origin. On average 13 species were detected in each bag. Tylospora fibrillosa, Amphinema sp., Tomentellopsis submollis, and Xerocomus badius made up almost 80% of the EM sequences. High biomass was related to increased dominance of specific species. There were no statistically significant differences in biomass production estimated from PLFA 18:2omega6, 9, or between fungal communities of apatite-amended and control bags estimated from DNA after one growth season. The potential of the mesh bag method in studies of functional diversity of EM mycelia in the field is discussed.

UNITE: a database providing web-based methods for the molecular identification of ectomycorrhizal fungi.

Identification of ectomycorrhizal (ECM) fungi is often achieved through comparisons of ribosomal DNA internal transcribed spacer (ITS) sequences with accessioned sequences deposited in public databases. A major problem encountered is that annotation of the sequences in these databases is not always complete or trustworthy. In order to overcome this deficiency, we report on UNITE, an open-access database. UNITE comprises well annotated fungal ITS sequences from well defined herbarium specimens that include full herbarium reference identification data, collector/source and ecological data. At present UNITE contains 758 ITS sequences from 455 species and 67 genera of ECM fungi. UNITE can be searched by taxon name, via sequence similarity using blastn, and via phylogenetic sequence identification using galaxie. Following implementation, galaxie performs a phylogenetic analysis of the query sequence after alignment either to pre-existing generic alignments, or to matches retrieved from a blast search on the UNITE data. It should be noted that the current version of UNITE is dedicated to the reliable identification of ECM fungi. The UNITE database is accessible through the URL http://unite.zbi.ee

Insect-like olfactory adaptations in the terrestrial giant robber crab.

The robber crab (Birgus latro), also known as the coconut crab, is the world's largest land-living arthropod, with a weight reaching 4 kg and a length of over half a meter. Apart from the marine larval stage, this crab is fully terrestrial, and will actually drown if submerged in water. A transition from sea to land raises dramatically new demands on the sensory equipment of an animal. In olfaction, the stimulus changes from hydrophilic molecules in aqueous solution to mainly hydrophobic in the gaseous phase. The olfactory system of land crabs thus represents an excellent opportunity for investigating the effects of the transition from sea to land. Have land crabs come to the same solutions as other terrestrial animals, or is their olfactory sense characterized by unique innovations? Here, we show that the robber crab has evolved an olfactory sense with a high degree of resemblance to the insect system. The similarities extend to physiological, behavioral, and morphological characters. The insect nose of the robber crab is a striking example of convergent evolution and nicely illustrates how similar selection pressures result in similar adaptation.

Divergence in gene expression related to variation in host specificity of an ectomycorrhizal fungus.

Ectomycorrhizae are formed by mutualistic interactions between fungi and the roots of woody plants. During symbiosis the two organisms exchange carbon and nutrients in a specific tissue that is formed at the contact between a compatible fungus and plant. There is considerable variation in the degree of host specificity among species and strains of ectomycorrhizal fungi. In this study, we have for the first time shown that this variation is associated with quantitative differences in gene expression, and with divergence in nucleotide sequences of symbiosis-regulated genes. Gene expression and sequence evolution were compared in different strains of the ectomycorrhizal fungus Paxillus involutus; the strains included Nau, which is not compatible with birch and poplar, and the two compatible strains Maj and ATCC200175. On a genomic level, Nau and Maj were very similar. The sequence identity was 98.9% in the 16 loci analysed, and only three out of 1075 genes analysed by microarray-based hybridizations had signals indicating differences in gene copy numbers. In contrast, 66 out of the 1075 genes were differentially expressed in Maj compared to Nau after contact with birch roots. Thirty-seven of these symbiosis-regulated genes were also differentially expressed in the ATCC strain. Comparative analysis of DNA sequences of the symbiosis-regulated genes in different strains showed that two of them have evolved at an enhanced rate in Nau. The sequence divergence can be explained by a decreased selection pressure, which in turn is determined by lower functional constraints on these proteins in Nau as compared to the compatible strains.

The influence of substrate pH on carbon translocation in ectomycorrhizal and non-mycorrhizal pine seedlings.

The effects of changed substrate pH on translocation and partitioning of 14 C-labeled plant assimilates were examined in laboratory microcosms containing mycorrhizal (unidentified fungal isolate 'Pink FMT 87:2') and non-mycorrhizal seedlings of Pinus sylvestris L. and Pinus contorta Dougl. ex Loud. The mycorrhizal plants had intact mycelial systems at different developmental stages, and microcosms contained non-sterile peat (pH 3.8) or peat adjusted to different pH values with CaO. In systems with mycorrhizal mycelium which had just started to colonize the peat no significant differences in 14 C assimilation were found, either with respect to substrate pH or mycorrhizal status of the plant. Loss of activity from the mycorrhizal plants was more rapid, however, probably mainly as a result of increased respiration from the infected root systems. After 8 wk growth in peat at pH 3.8 and 5.2 shoot weights of all seedlings were the same, whereas non-mycorrhizal plants had root systems twice the size of the mycorrhizal ones. In plants with well developed extramatrical mycelia translocation of labeled carbon to the mycelium growing at pH 3.8 was faster than that to mycelium growing at pH 5.2. After 4 d incubation, however, the percentage of the originally supplied carbon present in the mycelium was 5% regardless of substrate pH. Activity found in the peat surrounding non-mycorrhizal plants rarely exceeded 0.3%.

Effects of liming on ectomycorrhizal fungi infecting Pinus sylvestris L.: III. Saprophytic growth and host plant infection at different pH values in unsterile humus.

Five ectomycorrhizal fungi were studied in relation to the effects of applications of lime and wood ash to the growth substrate. Growth and survival of inoculated fungi in unsterile humus and relative root colonization frequency by inoculated and indigenous ectomycorrhizal fungi were measured. Growth of the fungi was tested in Petri dish systems with humus taken from a field site treated with lime and wood ash. The infection potential of the fungi was tested by introducing Pinus sylvestris L. seedlings into the growth systems. Paxillus involutus (Fr.) Fr. was the only fungus affected both by the pH increase, and by the different treatments applied, in all aspects of its ecology tested (growth and survival in humus, infection potential and competitive ability). No other fungus grew saprophytically, but they showed the similar changes in infection potential in response to pH whether lime or ash had been used.