Molecular phylogeny of the mycorrhizal desert truffles (Terfezia and Tirmania), host specificity and edaphic tolerance

Terfezia and Tirmania, so called desert truffles, are mycorrhizal fungi mostly endemic to arid and semi-arid areas of the Mediterranean Region, where they are associated with Helianthemum species. The aim of this work was to study the phylogenetic relationships in these pezizalean hypogeous fungi. The restriction fragment length polymorphism (RFLP) and DNA sequences of internal transcribed spacers (ITS) of the nuclear rDNA were studied for several morphological species, Terfezia arenaria, T. boudieri, T. claveryi, T. leptoderma, T. terfezioides (=Mattirolomyces terfezioides), Tirmania nivea and T. pinoyi. The sequences were analyzed with distance and parsimony methods. Phylogenetic analyses indicated a close genetic relationship between Tirmania and Terfezia. They may have arisen from a single evolutionary lineage of pezizalean fungi that developed the hypogeous habit as an adaptation to heat and drought in Mediterranean ecosystems. This analysis also supports the re-establishment of the genus Mattirolomyces. The genera Tirmania and Terfezia were monophyletic, and morphological species corresponded to phylogenetic species. The Tirmania clade comprises desert truffles with smooth spores and amyloid asci, which were found in deserts. The Terfezia clade grouped species found in semi-arid habitats having ornamented and spherical spores. These species are adapted to exploit different types of soil (either acid or basic soils) in association with specific hosts (either basophilous or acidophilous species). Although other factors might also play a role, host specialization and edaphic tolerances (fungus and/or host tolerances) might be the key in the species diversity of these genera.     

Fungus diversity in revegetated paddocks compared with remnant woodland in a south‐eastern Australian agricultural landscape

Summary Despite the importance of fungi for restoration, their presence in revegetated sites has received little attention. We compared the diversity and composition of macrofungi (i.e. those that form fleshy mushrooms and truffles) in 12 sites where 3‐to‐6‐year‐old native trees and shrubs had been planted (woodland restoration sites), with that in six woodland remnants. All sites were within an agricultural landscape near Holbrook in New South Wales. Of 58 fungal genera recorded, 57% occurred in woodland restoration sites and 83% in nearby patches of remnant woodland. Of the genera found in restoration sites, 70% were also found in the woodland remnants. The dominance of early successional genera such as Lacceria and Scleroderma in restoration sites suggests windblown colonisation by fungi. The reduced proportion of hypogeous genera (truffles) that rely on mammal vectors, which are less likely to occur in the restoration sites, also supports the view that most fungi occurred in restoration through colonisation rather than being generated from soil spores. Greatest overall fungal diversity occurred in large remnants that had greater structural complexity. Across all sites, epigeous genera (mushrooms) were most common (78% of all taxa collected across 46 genera) and of the nutritional modes, mycorrhizal genera (forming symbiotic associations with plants) were the most common (206 collections, 71%, 25 genera). Both hypogeous and mycorrhizal fungi were positively associated with the diversity of native forb species (wildflowers), suggesting that lower fungal diversity in restoration sites is likely to be a consequence of long‐term agricultural practices, particularly fertilizer use.     

Soil mycoflora from trufflè native areas of Saudi Arabia

Soil fungi of areas in the North-Eastern region of Saudi Arabia where truffles are native were surveyed. Forty-three species of fungi belonging to twenty genera were isolated. Most were recovered from soils underneath or around truffle ascocarps: thirty species from soil under the surface of Tirmania nivea ascocarps, twenty-four from Terfezia boudieri soil and twenty species each from Tirmania pinoyi and Terfezia claveryi soils. Rhizosphere soil of Helianthemum lippi, on the other hand, yielded twenty-four fungal species while only fourteen fungal species were found in soil without vegetation. The total counts of fungi/g soil were highest in soils from the under surface of truffles, followed by rhizosphere soil, with the lowest in soils without vegetation.     

Mites are the most common vectors of the fungus Gondwanamyces proteae in Protea infructescences

Entomochoric spore dispersal is well-documented for most ophiostomatoid fungal genera, most of which are associated with bark or ambrosia beetles. Gondwanamyces spp. are unusual members of this group that were first discovered in the flower heads of the primitive angiosperm genus Protea, that is mostly restricted to the Cape Floristic region of Africa. In this study, we present the discovery of the vectors of Gondwanamyces proteae in Protea repens infructescences, which were identified using PCR, direct isolation, and light microscopy. Gondwanamyces proteae DNA and ascospores were identified on diverse lineages of arthropods including beetles (Euderes lineicolis and Genuchus hottentottus), bugs (Oxycarenus maculates), a psocopteran species and five mite (Acari) species. Based on isolation frequency, however, a mite species in the genus Trichouropoda appears to be the most common vector of G. proteae. Gondwanamyces spores were frequently observed within pit mycangia at the base of the legs of these mites. Manipulative experiments demonstrated the ability of mites to carry viable G. proteae spores whilst in transit on the beetle G. hottentottus and that these mites are able to transfer G. proteae spores to uncolonised substrates in vitro. Interestingly, this same mite species has also been implicated as vector of Ophiostoma spores on P. repens and belongs to the same genus of mites that vector Ophiostoma spp. associated with pine-infesting bark beetles in the Northern Hemisphere.     

Low intensity surface fire instigates movement by adults of Calosoma frigidum (Coleoptera, Carabidae)

The genus Calosoma (Coleoptera: Carabidae) is a group of large, sometimes ornate beetles, which often voraciously attack caterpillars. Many studies have reported Calosoma beetles being highly conspicuous during defoliator outbreaks. Based on observations of individual beetle behavior, patterns of activity density and phenology we provide a hypothesis on how environmental cues may synchronize Calosoma activity with periods of high defoliation. We have observed that adults of Calosoma frigidum construct underground burrows similar to those reported to be created by larvae for pupation. We propose that small increases in soil surface temperature caused either by defoliation events or decreased albedo of blackened, burned soil causes beetles to leave their underground burrows and begin foraging. Indirect support for this hypothesis comes from high levels of adult Calosoma frigidum collected in relatively small patches of burned forest (<200m(2)) relative to the surrounding mosaic of unburned forest shortly after a prescribed surface burn.     

Diversity of pselaphine beetles (Coleoptera: Staphylinidae: Pselaphinae) in eastern Thailand

Pselaphine beetles (Coleoptera: Staphylinidae: Pselaphinae) are cosmopolitan, species‐rich, and yet poorly studied, particularly in the tropics. We sampled beetles in three types of primary forest and two types of disturbed forest habitats in eastern Thailand to assess the utility of pselaphine beetles as bioindicators of forest disturbance. We simultaneously measured leaf litter mass, soil moisture, soil acidity and canopy cover at each site to infer which environmental factors affect pselaphine beetle diversity and abundance. At each site, pselaphine beetles were extracted from ten 1 m² samples of leaf litter and soil with Tullgren funnels. We sampled 1867 adult beetles representing six supertribes, 51 genera and 114 morphospecies; 7% of the genera and 92% of the species were undescribed. Forest types differed significantly in species richness, abundance, diversity and evenness. Primary forest had greater numbers of species and individuals, and higher diversity indices (H′). Teak plantation and secondary forest had substantially fewer individuals and species of pselaphine beetles. Species composition differed between primary and degraded forests. Canopy cover, soil moisture, and leaf litter mass positively correlated with beetle species richness and abundance. Leaf litter mass and soil moisture were the two most important factors affecting the diversity of pselaphine beetle assemblages. Among the 114 morphospecies collected, 43 morphospecies were specific to two or three habitats and 64 morphospecies were found only in a single habitat. Thus pselaphine beetles appear to have rather narrow habitat requirements and their presence/absence was correlated with environmental differences. These traits make pselaphine beetles a suitable bioindicator taxon for assessing forest litter diversity and monitoring habitat change.     

Entomophthora lampyridarum,a fungal pathogen of the soldier beetle,Chauliognathus pennsylvanicus

In 1974 and 1979 in Clemson, South Carolina, adults of the common soldier beetle, Chauliognathus pennsylvanicus, were found to be infected by the fungal pathogen, Entomophthora lampyridarum. After infected beetles died they remained attached by their mandibles to foliage and flowers. The wings of infected beetles remained open, allowing conidiophores to develop on the upper surface of the abdomen. Primary conidia were elongate and measured 36.5 × 17.1 μm. Two types of secondary spores were formed: Type I spores were similar in form to primary conidia, but were somewhat smaller; Type II spores were formed at the apex of slender stalks and measured 37.7 × 15.3 μm. Resting spores were spherical, hyaline, and 22.3 μm in diameter.     

Miocene invertebrate trace fossils from a braided river environment,western Nebraska,U.S.A

Lower Miocene cross-stratified sands of the Gering and Monroe Creek Formations exposed on Scotts Bluff National Monument in western Nebraska, U.S.A., were deposited by migrating sand bars in a braided river system similar to the modern Platte River in eastern Nebraska and, like the Platte, contain local lenses of parallel stratified sediment that accumulated in ponded areas of abandoned channels. During times of low discharge, broad areas of river bar sands and abandoned channel sediments were subaerially exposed on the Miocene river plain. These sediments, like those exposed in the Platte River today, were subjected to burrowing by insects and other animals.Trace fossils in Lower Miocene braided river deposits are: vertical shelter burrows, horizontal deposit-feeding burrows, bioturbated layers, and vertical passageways between bioturbated layers. The burrows are cylindrical to sub-cylindrical in cross-section, internally meniscate or massive, generally non-branching, and smooth walled. Shelter burrows are similar in shape and size to recent burrows dug by beetles in river sediment for protection from day-time temperatures, to pass the night, and to hibernate. The shelter burrows, deposit-feeding burrows, and vertical passageways in the Lower Miocene sediments occur in four distinct “populations” with modal diameters of 1–2, 3–4, 7–8, and 10–12 mm. The occurrence of both vertical and horizontal burrows in all four “populations” suggests that they could have been made by the same insect. “Populations” with modal diameters of 1–2, 3–4, and 7–8 mm also occur in modern Platte River sediment and are made by tiger-beetle larvae (3–4 mm) and heterocerid (1–2 mm) beetles. Miocene shelter burrows, deposit-feeding burrows, bioturbated layers, and vertical passageways, therefore, could have been formed by different types of beetles, and/or larval instars and adults of the same beetle species.     

The diversity of Terfezia desert truffles: new species and a highly variable species complex with intrasporocarpic nrDNA ITS heterogeneity

Desert truffles belonging to Terfezia are well known mycorrhizal members of the mycota of the Mediterranean region and the Middle East. We aimed to test (i) whether the morphological criteria of Terfezia species regularly collected in Spain enable their separation and (ii) whether the previously hypothesized edaphic/biotic specificity of one group could be confirmed by study of a larger number of specimens. The species T. arenaria and T. claveryi can be identified unambiguously by morphological characters. We consider T. leptoderma as a distinct species while several lineages of similar spiny spored Terfezia truffles with cellular peridium were detected that have no obvious anatomical differences. Several species treated generally as synonyms of T. olbiensis have been described in this group, and because they cannot be unambiguously assigned to separate lineages we propose to consider the group as the T. olbiensis species complex. A high level of intrasporocarpic variation of the nrDNA ITS was detected in the T. olbiensis species complex, especially in one of its lineages. We detected no exclusive specificity to either plant associates or soil, except in T. leptoderma, which was associated with Quercus spp. and cistaceous plants on acidic soils. Nevertheless the clades showed a tendency either to associate with Quercus/Helianthemum/Cistus or Pinus hosts. Specimens having distinct anatomical features, reticulate spores and cellular peridium formed a separate group in the molecular phylogenetic analyses of nrDNA ITS and LSU regions; for these specimens we propose a new species, Terfezia alsheikhii sp. nov.     

Diversity and structure of AMF communities as affected by tillage in a temperate soil

Arbuscular mycorrhizal fungi (AMF) were studied in differently tilled soils from a long-term field experiment in Switzerland. Diversity and structure of AMF communities were surveyed either directly on spores isolated from the field soil or on spores isolated from trap cultures, planted with different host plants. Single-spore cultures were established from the AMF spores obtained from trap cultures. Identification of the AMF was made by observation of spore morphology and confirmed by sequencing of ITS rDNA. At least 17 recognised AMF species were identified in samples from field and/or trap cultures, belonging to five genera of AMF--Glomus, Gigaspora, Scutellospora, Acaulospora, and Entrophospora. Tillage had a significant influence on the sporulation of some species and non- Glomus AMF tended to be more abundant in the no-tilled soil. The community structure of AMF in the field soil was significantly affected by tillage treatment. However, no significant differences in AMF diversity were detected among different soil tillage treatments. AMF community composition in trap cultures was affected much more by the species of the trap plant than by the original tillage treatment of the field soil. The use of trap cultures for fungal diversity estimation in comparison with direct observation of field samples is discussed.     

Soil mycoflora from the Dead Sea Oases of Ein Gedi and Einot Zuqim (Israel)

Samples were taken from the top 10 cm of soils from 24 points in the Ein Gedi area. Among 329 isolates, 142 species were identified: 11 genera of ascomycetes, one genus of coelomycetes, 28 genera of hyphomycetes, 7 genera of zygomycetes and one yeast, in addition to some unidentified basidiomycetes. The hyphomycetes were represented by 17 dematiaceous, 9 mucedinaceous and two tuberculariaceous. Melanconiaceous and stilbellaceous genera were not found. Two new varieties of Microascus recently described were reisolated. No strict thermophiles or halophiles were obtained. There is apparently no very characteristic or specific fungal flora of the Dead Sea Oases although it was different from that found in the desert soil surrounding this area.     

Electrophysiological and behavioral responses of Dendroctonus frontalis (Coleoptera: Curculionidae) to volatiles isolated from conspecifics

Olfactory sensitivity of the southern pine beetle, Dendroctonus frontalis Zimmermann, to compounds isolated from the mid/hindguts of newly emerged conspecific adults was assayed with coupled gas chromatography-electroantennographic detection. All previously reported pheromones for D. frontalis plus eight additional compounds (fenchyl alcohol, myrtenal, cis-verbenol, trans-pinocarveol, acetophenone, trans-myrtanol, cis-myrtanol, and 2-phenylethanol) consistently elicited antennal responses from at least one sex. The eight additional compounds were assayed individually at three release rates (0.4-0.8, 3-9, and 25-100 mg/d) for the ability to alter D. frontalis responses to traps baited with D. frontalis attractant (4 mg/d frontalin and 17 mg/d alpha-pinene). At the high release rate, cis-verbenol enhanced attraction of D. frontalis females, whereas the other seven compounds significantly reduced attraction of one or both sexes. Acetophenone significantly reduced attraction of male D. frontalis at the low release rate, and five compounds (fenchyl alcohol, trans-pinocarveol, acetophenone, cis-myrtanol, and 2-phenylethanol) reduced attraction of one or both sexes at the intermediate rate. Only acetophenone significantly altered the sex ratio of beetles trapped, decreasing the proportion of males. Attraction of predatory checkered beetles (Cleridae) was enhanced by cis-verbenol released at the high rate but was not altered by any compound inhibitory to D. frontalis. Analyses of volatiles from individual D. frontalis indicated that the majority of the eight compounds were produced in greater quantities by newly emerged beetles than ones attacking pine bolts. Five of the compounds were associated predominantly with one sex. Possible ecological roles of these compounds in the biology of D. frontalis are discussed.     

The action of post-dispersal beetles (Coleoptera: Scarabaeidae) and ants (Hymenoptera: Formicidae) on scats of Didelphis spp. (Mammalia: Didelphidae)

A two year study of dung beetles and ants acting on scats of two species of opossum (Didelphis spp.) was carried out. Scats were left in the field in order to detect post-dispersal agents. A portion of each scat (30 %) was examined for seeds in the laboratory. Beetles were recovered from burrows (51% of 84 faecal samples left in the field) where they either buried scats of opossums or were attracted, together with ants, to pitfalls (N = 10) baited with opossum scats. Dung beetles were the main post-dispersal agents of seeds found in scats of opossums, rolling the scats away or burying then on the site of deposition. They buried faeces at 4 to 15 cm in depth (N = 22 tunnels). The main dung beetles identified (medium to large size) were Eurysternus (28.7 % in pitfalls) and Dichotomius (13.7 %), Coprophanaeus (seen only directly on faeces), besides small-bodied beetles (< 10 mm; 57.6 %). The ant Acromirmex sp. transported some seeds from scats. This species was present in 25.5 % of all Formicidae samples (pitfall). These post-dispersal agents contribute to avert scat seed predators such as rodents, and to accelerate seed bank formation.     

Desert Truffles of the Australian Outback: Ecology,Ethnomycology, and Taxonomy

Desert Truffles of the Australian Outback: Ecology, Ethnomycology, and Taxonomy. The Aborigines of central Australia have traditionally used desert truffles as food. Truffle hunting in the desert requires substantial ecological knowledge, as truffles occur sporadically and only with adequate and properly distributed rainfall as well as the presence of necessary soil conditions and mycorrhizal hosts. Truffles are hunted primarily by women, who look for cracks or humps in the soil caused by expansion of the truffles, which are then extracted with digging sticks. The truffles are typically eaten raw or baked or roasted in ashes. Seven truffle species are recorded from the Australian Outback, including three that have been only recently described.     

Olfactory orientation of the truffle beetle, Leiodes cinnamomea

Although the truffle beetle, Leiodes cinnamomea, inflicts substantial damage to the ripe stage of fruiting bodies of the economically important black truffle (Tuber melanosporum), it is not attracted by ripe truffle odours. Rather, male beetles are attracted to infested truffles only in the presence of female beetles, suggesting that the former employ a pheromone to locate truffles over short distances. In contrast, female beetles show no attraction to infested or uninfested truffles, suggesting that they employ other cues, possibly linked to odours emitted by truffles prior to the ripe stage. We hypothesize that the chemical composition of truffle volatiles changes over the life of the truffle fruiting body, being attractive to insects early on and to mammals just prior to decomposition.     

Attraction of the southern pine beetle, Dendroctonus frontalis, to pheromone components of the western pine beetle, Dendroctonus brevicomis (Coleoptera: Curculionidae: Scolytinae), in an allopatric zone

Subtle differences in pheromone components of sympatric species should be attractive only to the producing species and unattractive or repellent to the nonproducing species, and thereby maintain reproductive isolation and reduce competition between species. Bark beetles Dendroctonus brevicomis and D. frontalis (Coleoptera: Curculionidae) are known to have common pheromone components, except for exo-brevicomin, which is produced by D. brevicomis. We predicted that D. frontalis would not respond to exo-brevicomin outside of the zone of sympatry with D. brevicomis. We conducted a field experiment to determine the effect of exo-brevicomin on attraction of D. frontalis and associated species in Mississippi. We determined whether D. frontalis pheromone production differed inside and outside the sympatric zone and compared the pheromone profiles with D. brevicomis within the sympatric zone. Trapping studies revealed that D. frontalis can perceive and respond positively to exo-brevicomin, an aggregation pheromone of a sympatric congener (D. brevicomis), at locations hundreds of kilometers outside the sympatric zone. Qualitative pheromone profiles showed that both species emit similar pheromone components: frontalin, endo-brevicomin, exo-brevicomin, trans-verbenol, verbenone, and myrtenol. Although not previously reported, D. frontalis males from Arizona produced exo-brevicomin. The predator Thanasimus dubius did not discriminate traps baited with exo-brevicomin and was most attracted to traps with frontalin. Hylastes beetles were significantly attracted to traps baited with exo-brevicomin in combination with other compounds. Our results raise new practical and evolutionary questions on the role of exo-brevicomin in the behavioral ecology of D. frontalis. The addition of exo-brevicomin to the current lure might increase the efficiency of trapping programs in the southeastern United States.     

Deforestation and biodiversity: effects of bushland cultivation on dung beetles in semi-arid Tanzania

Dung beetles provide important ecosystem services in the habitats where they occur. The activity of dung beetles enhances soil nutrient cycling and increases the soil’s ability to absorb and hold water. Consequently, these beetles are particularly important in semi-arid environments. This study analyses the importance of remaining wooded habitat patches (bushland) for the survival of a diverse dung beetle fauna in an otherwise cultivated landscape in semi-arid Tanzania. Dung beetles were sampled by pitfall trapping in maize fields and bushland habitats. In total, 6037 dung beetles (Scarabaeidae: Scarabaeinae), representing 77 species from 25 genera, were collected. Many species, particularly amongst the ball-rollers, showed a clear preference for wooded patches, species richness being significantly higher in the bushland patches than in the cultivated sites. The number of trapped specimens in bushland was also considerably higher than that found in maize fields, although the differences were not significant. In conclusion, bushland fragments appear to have an important conservation value as to maintaining a high diversity and abundance of dung beetles, thereby enhancing the ecosystem services provided by these beneficial insects.     

西双版纳热带雨林中丛枝菌根真菌的初步研究*

对西双版纳热带雨林中30个科的42种植物根系的丛枝菌根真菌定居情况进行了调查,并从这些植物的根际土壤中分离鉴定了分属于无梗囊霉属(Acaulospora)、球囊霉属(Glomus)和硬囊霉属(Sclerocystis)的25种丛枝菌根真菌。对热带雨林土壤中丛枝菌根真菌的孢子密度（spore density）、物种丰富度（species richness）以及已鉴定种的出现频率进行统计分析发现：热带雨林土壤中丛枝菌根真菌的孢子密度在每100g土壤116~1560个之间,平均478个;物种丰富度在2~7之间,平均为4.5;无梗囊霉属和球囊霉属真菌是热带雨林土壤中丛枝菌根真菌的优势类群。     

Population traits of the burrowing toad Rhinella fernandezae (Gallardo, 1957) (Anura, Bufonidae).

Size distribution, sex ratio and use of burrows of the burrowing toad Rhinella fernandezae were studied in Buenos Aires province, Argentina. Two sites separated by approximately 300 m were studied: one was a road next to a swamp, and the other a garden of a country house located further from the swamp. We identified toad burrows, and individuals were sexed, measured and given an individual mark. Burrows were examined in subsequent months after the first sampling to assess the presence of toads. We found significant differences in the size distribution between areas, being the proportion of juveniles greater at the site next to the swamp where the reproduction of the species was observed. This result may suggest that the site located near to the swamp functions as a source habitat of individuals that migrate to the other site, where recruitment would be very scarce. Sex proportion of adults did not differ from 1:1 in neither the total population nor in each site, suggesting that there was not differential mortality by sex. Some toads changed burrows throughout the study period, but there were not differences in the frequency of change between adults and juveniles.