Coprinopsis cinerea dioxygenase is an oxygenase forming 10(S)-hydroperoxide of linoleic acid, essential for mushroom alcohol, 1-octen-3-ol, synthesis.

1-Octen-3-ol is a volatile oxylipin found ubiquitously in Basidiomycota and Ascomycota. The biosynthetic pathway forming 1-octen-3-ol from linoleic acid via the linoleic acid 10(S)-hydroperoxide was characterized 40 years ago in mushrooms, yet the enzymes involved are not identified. The dioxygenase 1 and 2 genes (Ccdox1 and Ccdox2) in the mushroom Coprinopsis cinerea contain an N-terminal cyclooxygenase-like heme peroxidase domain and a C-terminal cytochrome P450-related domain. Herein, we show that recombinant CcDOX1 is responsible for dioxygenation of linoleic acid to form the 10(S)-hydroperoxide, the first step in 1-octen-3-ol synthesis, whereas CcDOX2 conceivably forms linoleic acid 8-hydroperoxide. We demonstrate that KO of the Ccdox1 gene suppressed 1-octen-3-ol synthesis, although added linoleic acid 10(S)-hydroperoxide was still efficiently converted. The P450-related domain of CcDOX1 lacks the characteristic Cys heme ligand and the evidence indicates that a second uncharacterized enzyme converts the 10(S)-hydroperoxide to 1-octen-3-ol. Additionally, we determined the gene KO strain (ΔCcdox1) was less attractive to fruit fly larvae, while the feeding behavior of fungus gnats on ΔCcdox1 mycelia showed little difference from that on the mycelia of the WT strain. The proliferation of fungivorous nematodes on ΔCcdox1 mycelia was similar to or slightly worse than that on WT mycelia. Thus, 1-octen-3-ol seems to be an attractive compound involved in emitter-receiver ecological communication in mushrooms.

Pre-pollination barriers between two sympatric Arisaema species in northern Shikoku Island, Japan.

PREMISE: The genus Arisaema (Araceae) has rapidly diversified in Japan, and multiple species often coexist in the field. Although Japanese Arisaema species hybridize from artificial crossing, hybrid individuals are rare in mixed populations; suggesting the presence of effective pre-pollination barriers. We examined the following reproductive barriers between A. sikokianum and A. tosaense: habitat, phenology, and pollinator isolations. METHODS: Habitat isolation was examined by interspecific comparisons of microhabitat conditions at a mixed site and of altitude at the sampling site of herbarium specimens. Phenological isolation was evaluated by comparing seasonal transition in apparent spathe condition and frequency of insect visitation. Pollinator isolation was examined by comparing floral visitor assemblages between the two Arisaema species. To avoid overestimation of pollinator isolation due to seasonal changes in insect assemblages, we also compared visitor assemblages between natural and late-flowering A. sikokianum, where the latter was experimentally introduced and blooming with a natural A. tosaense population. RESULTS: Microhabitat conditions and sampling elevations of herbarium specimens overlapped between the two Arisaema species. At the population level, A. sikokianum and A. tosaense flowered for 39 and 52 days, respectively, with 13 days overlap. Insect visitation in A. sikokianum decreased before the seasonal overlap. Floral visitor assemblages differed between the two Arisaema species, while the difference between natural and late-flowering A. sikokianum was less distinct. CONCLUSIONS: Phenological and pollinator isolation contribute to reproductive isolation between the two Arisaema species and should enable the two species to coexist in this area.

Priority effects are interactively regulated by top-down and bottom-up forces: evidence from wood decomposer communities.

Both top-down (grazing) and bottom-up (resource availability) forces can determine the strength of priority effects, or the effects of species arrival history on the structure and function of ecological communities, but their combined influences remain unresolved. To test for such influences, we assembled experimental communities of wood-decomposing fungi using a factorial manipulation of fungivore (Folsomia candida) presence, nitrogen availability, and fungal assembly history. We found interactive effects of all three factors on fungal species composition and wood decomposition 1 year after the fungi were introduced. The strength of priority effects on community structure was affected primarily by nitrogen availability, whereas the strength of priority effects on decomposition rate was interactively regulated by nitrogen and fungivores. These results demonstrate that top-down and bottom-up forces jointly determine how strongly assembly history affects community structure and function.

Examining the Shift in the Decomposition Channel Structure of the Soil Decomposer Food Web: A Methods Comparison.

Selecting the appropriate indicators and measuring time point numbers is important for accurately examining the shift in soil gross decomposition channel structure. Through a selected case study on a natural forest vs. rainfed arable system over a two-month-long experiment, the utility of three commonly employed indicators (fungi to bacteria ratio (F:B), fungivore to bacterivore ratio (FF:BF), and glucosamine to muramic acid ratio (GlcN:MurN)) were compared to reflect the shift in soil gross decomposition channel structure. The requirement of measuring the time point numbers for the three indicators was also assessed, and we suggest a potential methodology. Our results revealed that the GlcN:MurN ratio was more reliable for assessing the shifts in gross decomposition channel structure for long-term land use changes, while it was less sensitive to short-term drought compared with the other two indicators. The F:B ratio was more applicable than the FF:BF ratio for reflecting both long- and short-term changes. Furthermore, the reliability of the GlcN:MurN ratio was the least dependent on measuring time point numbers. We suggest the use of multiple indicators and the adoption of multiple measuring time points for the overall methodology.

Activation of mycelial defense mechanisms in the oyster mushroom Pleurotus ostreatus induced by Tyrophagus putrescentiae.

Fungal chemicals are vital in processes recognizing damage- and microbe-associated molecules (DAMPs/MAMPs) that trigger defense responses in fungi. Pleurotus ostreatus is a widely cultivated edible fungus that is prone to attack from fungivorous insects and mites. Yet P. ostreatus has evolved an elegant defense system against fungivore attacks. In this study, we investigated how the oyster mushroom responds to the fungivory and mechanical wounding by conducting transcriptome, proteome, and secondary metabolic analyses. The profiling analysis revealed a total of 11,495 transcripts and 866 proteins, 4416 differentially expressed genes (DEGs), and 62 differentially expressed proteins (DEPs) were identified in response to the mite Tyrophagus putrescentiae feeding and mechanical wounding. In comparing the responses induced by mechanical wounding, some genes, proteins, and metabolites were uniquely induced or repressed by the mite. At the transcript level, nine pathways were activated by the mite feeding, including those of "MAPK signaling pathway-yeast", "Phenylalanine metabolism", and "Biotin metabolism", among others, while both enrichment of "Ribosome", "Ribosome biogenesis in eukaryotes", and "Regulation of Mitophagy in Yeast" demonstrated the common effects upon fungal secretory protein synthesis and processing induced by fungivory and mechanical wounding. Fungivory also stimulated the synthesis of C8-aryl compounds and sesquiterpenes (especially1-octen-3-ol and α-/ß-bisabolene), and these compounds repellent to T. putrescentiae. Both jasmonic acid (JA) and jasmonic acid methyl ester (MeJA) were specifically regulated by mite feeding and mechanical wounding. The terpene synthase gene transcription was significantly increased induced by the exogenous addition of MeJA, resulting in defensive sesquiterpene production against the mite. These findings are the first to demonstrate that the reactive oxygen species (ROS)/MAPK signaling pathway, JA regulation, specific gene expression, and protein synthesis, and anti-mite substance metabolism are all involved in coordinated inducible chemical-based defense responses in P. ostreatus, which could be especially effective the mite T. putrescentiae.

Characterization of Microbial Communities from the Alimentary Canal of Typhaea stercorea (L.) (Coleoptera: Mycetophagidae).

The gut microbiomes of symbiotic insects typically mediate essential functions lacking in their hosts. Here, we describe the composition of microbes residing in the alimentary canal of the hairy fungus beetle, Typhaea stercorea (L.), at various life stages. This beetle is a post-harvest pest of stored grains that feeds on fungi and serves as a vector of mycotoxigenic fungi. It has been reported that the bacterial communities found in most insects' alimentary canals contribute to nutrition, immune defenses, and protection from pathogens. Hence, bacterial symbionts may play a key role in the digestive system of T. stercorea. Using 16S rRNA amplicon sequencing, we examined the microbiota of T. stercorea. We found no difference in bacterial species richness between larvae and adults, but there were compositional differences across life stages (PERMANOVA:pseudo-F(8,2) = 8.22; p = 0.026). The three most abundant bacteria found in the alimentary canal of the larvae and adults included Pseudomonas (47.67% and 0.21%, respectively), an unspecified genus of the Enterobacteriaceae family (46.60 % and 90.97%, respectively), and Enterobacter (3.89% and 5.75%, respectively). Furthermore, Pseudomonas spp. are the predominant bacteria in the larval stage. Our data indicated that field-collected T. stercorea tended to have lower species richness than laboratory-reared beetles (Shannon: H = 5.72; p = 0.057). Furthermore, the microbial communities of laboratory-reared insects resembled one another, whereas field-collected adults exhibited variability (PERMANOVA:pseudo-F(10,3) = 4.41; p = 0.006). We provide evidence that the environment and physiology can shift the microbial composition in the alimentary canal of T. stercorea.

Effects of enhanced productivity of resources shared by predators in a food-web module: Comparing results of a field experiment to predictions of mathematical models of intra-guild predation.

We compared the response to resource enhancement of a simple empirical model of intra-guild predation (IGP) to the predictions of published, simple mathematical models of asymmetric IGP (a generalist IG Predator that feeds both on a specialist IG Prey and a Resource that it shares with the IG Prey). The empirical model was a food-web module created by pooling species abundances across many families in a speciose community of soil micro-arthropods into three categories: IG Predator (large predatory mites), IG Prey (small predatory mites), and a shared Resource (fungivorous mites and springtails). By pooling abundances of species belonging to broadly defined functional groups, we tested the hypothesis that IGP is a dominant organizing principle in this community. Simple mathematical models of asymmetric IGP predict that increased input of nutrients and energy to the shared Resource will increase the equilibrium density of Resource and IG Predator, but will decrease that of IG Prey. In a field experiment, we observed how the three categories of the empirical model responded to two rates of addition of artificial detritus, which enhanced the food of fungivores, the Resource of the IGP module. By the experiment's end, fungivore densities had increased ~1.5× (ratio of pooled fungivore densities in the higher-input treatment to plots with no addition of detritus), and densities of IG Predators had increased ~4×. Contrary to the prediction of mathematical models, IG Prey had not decreased, but instead had increased ~1.5×. We discuss possible reasons for the failure of the empirical model to agree with IGP theory. We then explore analogies between the behavior of the empirical model and another mathematical model of trophic interactions as one way to gain insights into the trophic connections in this community. We also propose one way forward for reporting comparisons of simple empirical and mathematical models.

Natural history of the social millipede Brachycybe lecontii Wood, 1864.

The millipede Brachycybe lecontii Wood, 1864 is a fungivorous social millipede known for paternal care of eggs and forming multi-generational aggregations. We investigated the life history, paternal care, chemical defence, feeding and social behaviour of B. lecontii and provided morphological and anatomical descriptions, using light and scanning electron microscopy. Based on observations of B. lecontii from 13 locations throughout its distribution, we report the following natural history aspects. The oviposition period of B. lecontii lasted from mid-April to late June and the incubation period lasted 3-4 weeks. Only males cared for the eggs and subsequent care of juveniles was not observed. In one case, the clutches of two males became combined and they were later cared for by only one of the males. The defensive compound of B. lecontii is stored in large glands occupying a third of the paranotal volume and were observed only in stadia II millipedes and older. We observed B. lecontii feeding on fungi of the order Polyporales and describe a cuticular structure on the tip of the labrum that may relate to fungivory. We found that their stellate-shaped aggregations (pinwheels) do not form in the absence of fungus and suggest the aggregation is associated with feeding. We describe and illustrate a previously undescribed comb-like structure on the tibia and tarsi of the six anterior-most leg-pairs and measure the colour and spectral reflectance of the B. lecontii exoskeleton.

Mystery mushroom malingerers: Megaselia marquezi Hartop et al. 2015 (Diptera: Phoridae).

A mysterious female phorid fly, known for many years to be associated with fungal sporophores ("mushrooms") is identified as Megaselia marquezi Hartop et al. 2015. Male and female flies were collected emerging from the fungus Psathyrella candolleana (Fr.) Maire, and females were observed swarming over the sporophores.

First record of Neoempheria Osten Sacken (Diptera, Mycetophilidae) biology in the Neotropical region, with associations between its larvae and fungi.

BACKGROUND: Members of the family Mycetophilidae (Diptera) have life cycles that are typically associated with fungus. Their biology is relatively well known in the Palaearctic, though other regions are poorly known, and there are no associations recorded between mycetophilid immatures and fungi in the Neotropical region. Here we report the first association between a mycetophilid-Neoempheriapuncticoxa Edwards-and fungi in this region. Immatures of N.puncticoxa were collected on fungi and some were reared in the laboratory until adult emergence. The immature stages and adult of N.puncticoxa are described and re-described respectively, and high resolution images and illustrations of the habitus, wings, thorax, male and female terminalia, immatures, and in situ specimens are given. NEW INFORMATION: We report the first association between Mycetophilidae and fungi in the Neotropical region.

Patchiness and compensatory growth in a fungus-Collembola system.

The compensatory growth potential of a grazed fungal biomass was mathematically expressed as a function of patchiness in its distribution and demonstrated in an experiment using the fungivorous collembolan Onychiurus armatus and the soil fungi Verticillium bulbillosum and Penicillium spinulosum. The model addresses the regrowth potential in relation to patch fragmentation, travelling time and consumption rate of the collembolan and the mean relative growth rate of the fungus. It suggests that the mean relative growth rate required for regrowth decreases with patch fragmentation and increases with the mean growth rate of the fungus. The experiments were performed with a system of soil-filled vials provided with fungi and collembolans. The size of the vials and the length of the tubes connecting them were varied to give different patch sizes and travelling times. The respiratory activity of fungi after grazing increased as a unit of mycelium was distributed into smaller connected vials. The slow growing species V. bulbillosum showed a greater but delayed response to grazing in comparison with the fast growing P. spinulosum. An increased travelling time delayed the growth response in both species.

Nematode trophic structure in conventional and no-tillage agroecosystems.

The effect of tillage intensity on nematode community trophic structure and the role of nematodes in the regulation of decomposition rates in agroecosystems were examined. Conventional (CT) and no-tillage (NT) agroecosystems were sampled monthly for 1 year. Tillage affected nematode trophic structure and total abundance. Monthly mean densities of bacterivorous, fungivorous, and total nematodes were greater in CT than in NT plots. In the summer, however, fungivorous and plant parasitic nematodes were more abundant in NT. No difference was detected for omnivore-predator nematodes.