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Soil fungal diversity was studied by next-generation sequencing and compared in two different Malagasy ecosystems, the first a New Protected Area (Maromizaha NAP) that is a rich humid evergreen forest and the second a degraded and declined deciduous forest (Andaravina) whose area has been also eroded. Both areas, however, have comparable annual rainfalls and soil pH values. So it was of interest to examine the soil fungal diversity in each system and compare them. We detected 1,817,658 reads representing Ascomycota, which were dominant in both habitats (55.9%), followed by unidentified fungi (21.5%), Basidiomycota (12.7%) and Mortierellomycota (6.7%), with Mucoromycota, Chytridiomycota, Glomeromycota and other phyla accounting for less than 5% in total. In detail, 1,142 OTUs out of 1,368 constitute the common core shared by both sampling areas, which are characterized by tropical climate, whereas 185 are Maromizaha specific and 41 Andaravina specific. The most represented guilds involve fungi related to saprotrophic behaviour, with a greater tendency towards pathotrophic mode. A significant variability in terms of richness and abundance is present within Maromizaha, which is a heterogeneous environment for fungi but also for plant composition, as it emerged from the vegetational survey of the investigated sites. A few fungal sequences match taxa from Madagascar, highlighting the scarce representativeness of fungi from this island in the fungal databases and their still low knowledge. Enlarging studies in Madagascar will help not only to unravel its largely unknown fungal biodiversity but also to give a contribution for studies on the reconstruction of the diversity of soil fungi worldwide.

Nowadays, the worldwide agriculture is experiencing a transition process toward more sustainable production, which requires the reduction of chemical inputs and the preservation of microbiomes’ richness and biodiversity. Plants are no longer considered as standalone entities, and the future of agriculture should be grounded on the study of plant-associated microorganisms and all their potentiality. Moreover, due to the climate change scenario and the resulting rising incidence of abiotic stresses, an innovative and environmentally friendly technique in agroecosystem management is required to support plants in facing hostile environments. Plant-associated microorganisms have shown a great attitude as a promising tool to improve agriculture sustainability and to deal with harsh environments. Several studies were carried out in recent years looking for some beneficial plant-associated microbes and, on the basis of them, it is evident that Actinomycetes and arbuscular mycorrhizal fungi (AMF) have shown a considerable number of positive effects on plants’ fitness and health. Given the potential of these microorganisms and the effects of climate change, this review will be focused on their ability to support the plant during the interaction with abiotic stresses and on multi-omics techniques which can support researchers in unearthing the hidden world of plant–microbiome interactions. These associated microorganisms can increase plants’ endurance of abiotic stresses through several mechanisms, such as growth-promoting traits or priming-mediated stress tolerance. Using a multi-omics approach, it will be possible to deepen these mechanisms and the dynamic of belowground microbiomes, gaining fundamental information to exploit them as staunch allies and innovative weapons against crop abiotic enemies threatening crops in the ongoing global climate change context.

The development of biotechnologies based on beneficial microorganisms for improving soil fertility and crop yields could help to address many current agriculture challenges, such as food security, climate change, pest control, soil depletion while decreasing the use of chemical fertilizers and pesticides. Plant growth-promoting (PGP) microbes can be used as probiotics in order to increase plant tolerance/resistance to abiotic/biotic stresses and in this context strains belonging to the Pseudomonas chlororaphis group have shown to have potential as PGP candidates. In this study a new P. chlororaphis isolate is reported and tested for (i) in vitro PGP features, (ii) whole-genome sequence analysis, and (iii) its effects on the rhizosphere microbiota composition, plant growth, and different plant genes expression levels in greenhouse experiments. Results showed that P. chlororaphis ST9 is an efficient rice root colonizer which integrates into the plant resident-microbiota and affects the expression of several plant genes. The potential use of this P. chlororaphis strain as a plant probiotic is discussed.

Plant specialized metabolites (SMs) play an important role in the interaction with the environment and are part of the plant defense response. These natural products are volatile, semi-volatile and non-volatile compounds produced from common building blocks deriving from primary metabolic pathways and rapidly evolved to allow a better adaptation of plants to environmental cues. Specialized metabolites include terpenes, flavonoids, alkaloids, glucosinolates, tannins, resins, etc. that can be used as phytochemicals, food additives, flavoring agents and pharmaceutical compounds. This review will be focused on Mediterranean crop plants as a source of SMs, with a special attention on the strategies that can be used to modulate their production, including abiotic stresses, interaction with beneficial soil microorganisms and novel genetic approaches.

Plant essential oils (EO) possess antifungal properties due to the presence of terpenoids and phenolics capable of damaging microbial cell membranes. The compounds present in EO could represent a valid alternative to synthetic fungicides in rice crop protection, especially in Europe, where the available plant protection products are increasingly scarce, particularly for managing seed-borne fungal pathogens like Fusarium fujikuroi Nirenberg, the causal agent of bakanae disease. Bakanae is a major concern in Europe for seed producers, which often fail to comply with the strict regulatory limits on the presence of the pathogen in the field, resulting in the rejection of the seed crop from marketing and economic losses. The present work was aimed at studying 1) the in vitro effect of EO derived compounds against F. fujikuroi and 2) their efficacy in disease management under field conditions applied as seed treatments. Results confirmed the in vitro antifungal activity of tested compounds: specifically, thymol (0.025% vol/vol), terpinen-4-ol (0.1%), and eugenol (0.05%) showed strong antifungal activity against all the tested F. fujikuroi strains. Two-years, two-locations open field trials showed that seeds treated with thymol (20%) and eugenol (5%) significantly reduced bakanae incidence compared to untreated control, without any phytotoxicity effect on rice seed. The efficacy of these two compounds in managing the disease without affecting germination rates was comparable to the commercially available prochloraz and triticonazole formulation, considered as the positive control. Our results suggest that thymol and eugenol could be used as alternative fungicides in innovative formulations for seed dressing, with the advantage of being compliant with the European IPM framework and organic farming legislation.

The development of biotechnologies based on beneficial microorganisms for improving soil fertility and crop yields could help addressing many current agriculture challenges, such as food security, climate change, pests control, soil depletion while decreasing the use of chemical fertilizers and pesticides. Plant Growth Promoting (PGP) microbes can be used as probiotics in order to increase plant tolerance/resistance to abiotic/biotic stresses and in this context strains belonging to thePseudomonas chlororaphisgroup have shown to have potential as PGP candidates. In this work a newP. chlororaphisisolate is reported and tested for (i)in vitroPGP features, (ii) whole genome sequence analysis, and (iii) its effects on root microbiome, plant growth and on the expression of different plant genes in greenhouse experiments. The potential use of thisP. chlororaphisstrain as a plant probiotic is discussed.

The aim of this opinion is not only to report the low percentage of dataset related to the soil metaproteomic studies, but also to point out the still concealed potential of the technique if flanked by a proper repository of data.

Background Rice blast, caused by the fungus Pyricularia oryzae, represents the most damaging fungal disease of rice worldwide. Utilization of rice resistant cultivars represents a practical way to control the disease. Most of the rice varieties cultivated in Europe and several other temperate regions are severely depleted of blast resistance genes, making the identification of resistant sources in genetic background adapted to temperate environments a priority. Given these assumptions, a Genome Wide Association Study (GWAS) for rice blast resistance was undertaken using a panel of 311 temperate/tropical japonica and indica accessions adapted to temperate conditions and genotyped with 37,423 SNP markers. The panel was evaluated for blast resistance in field, under the pressure of the natural blast population, and in growth chamber, using a mixture of three different fungal strains. Results The parallel screening identified 11 accessions showing high levels of resistance in the two conditions, representing potential donors of resistance sources harbored in rice genotypes adapted to temperate conditions. A general higher resistance level was observed in tropical japonica and indica with respect to temperate japonica varieties. The GWAS identified 14 Marker-Traits Associations (MTAs), 8 of which discovered under field conditions and 6 under growth chamber screening. Three MTAs were identified in both conditions; five MTAs were specifically detected under field conditions while three for the growth chamber inoculation. Comparative analysis of physical/genetic positions of the MTAs showed that most of them were positionally-related with cloned or mapped blast resistance genes or with candidate genes whose functions were compatible for conferring pathogen resistance. However, for three MTAs, indicated as BRF10, BRF11–2 and BRGC11–3, no obvious candidate genes or positional relationships with blast resistance QTLs were identified, raising the possibility that they represent new sources of blast resistance. Conclusions We identified 14 MTAs for blast resistance using both field and growth chamber screenings. A total of 11 accessions showing high levels of resistance in both conditions were discovered. Combinations of loci conferring blast resistance were identified in rice accessions adapted to temperate conditions, thus allowing the genetic dissection of affordable resistances present in the panel. The obtained information will provide useful bases for both resistance breeding and further characterization of the highlighted resistance loci.

The completeness and accuracy of the reports of suspected adverse drug reactions is important in pharmacovigilance. The aim of the present study was to analyze the quality of the information included in the reports sent to the Pharmacovigilance Centre of São Paulo (Brazil).A sample of 999 reports received from January 2013 to December 2014 was selected. The quality of the filled information was evaluated according to a 'sufficiency' criterion to apply the Karch-Lasagna causality algorithm.There were 820 reports from manufacturers and 179 from health centres. Only 4.4% (44) were fully filled, thus allowing the adequate analysis of the causal relationship between the suspected medication and the adverse event. In 30% of the reports from manufacturers, the information about the critical variables was lacking or incomplete, preventing the adequate evaluation of the report. It was also noted that the reports' poor filling quality was not related with less severity or with old and well-known medicines.The poor quality of the information included in the reports received by this centre, especially those sent by pharmaceutical manufacturers, hampers the identification of potential safety signals. Measures to improve the quality of the reports must be urgently adopted.

Several important bacterial characteristics, such as biological nitrogen fixation, phosphate solubilization, 1‐aminocyclopropane‐1‐carboxylate deaminase activity and production of siderophores and phytohormones, can be assessed as plant growth promotion traits. Our aim was to evaluate the effects of nitrogen fixing and indole‐3‐acetic acid (IAA) producing endophytes in two Oryza sativa cultivars (Baldo and Vialone Nano). Three bacteria, Herbaspirillum huttiense RCA24, Enterobacter asburiae RCA23 and Staphylococcus sp. 377, producing different IAA levels, were tested for their ability to enhance nifH gene expression and nitrogenase activity in Enterobacter cloacae RCA25. Results showed that H. huttiense RCA24 performed best. Improvement in nitrogen fixation and changes in physiological parameters such as chlorophyll, nitrogen content and shoot dry weight were observed for plants co‐inoculated with strains RCA25 and RCA24 in a 10:1 ratio. Based on confocal laser scanning microscopy analysis, strain RCA24 was the best colonizer of the root interior and the only IAA producer located in the same root niche occupied by RCA25 cells. This work shows that the choice of a bio‐inoculum having the right composition is one of the key aspects to be considered for the inoculation of a specific host plant cultivar with microbial consortia., The contribution of IAS‐CSIC was supported by grant P12‐AGR‐667 (Junta de Andalucía, Spain), co‐funded by ERDF of the EU. The contribution of CREA‐CI was realized within the framework of Risobiosystems project (DM n. 94667, MiPAAFT), funded by MIPAAFT.

The role of ectomycorrhizal fungi in modulating host susceptibility to fungal pathogens is poorly known. Pinus sylvestris susceptibility to two congeneric fungal pathogens was compared between mycorrhizal and non-mycorrhizal plants. The ectomycorrhizal fungus was Suillus luteus, while the pathogens were Heterobasidion irregulare and H. annosum. H. irregulare is native to North-America and invasive to Europe, whereas S. luteus and H. annosum are native to Eurasia. Non-mycorrhizal plants were equally susceptible to both pathogens. Mycorrhizal plants were significantly less susceptible to H. annosum than non-mycorrhizal plants, whereas there were no differences when considering H. irregulare. The abundance of ectomycorrhizas was negatively and significantly correlated to the level of host susceptibility only to H. annosum. This study shows that the protective role of ectomycorrhizal fungi may differ when the host is challenged by congeneric fungal pathogens. Results add a further clue to explain and predict the invasiveness of H. irregulare in Europe. (C) 2018 Elsevier Ltd and British Mycological Society. All rights reserved. This work was supported by the Italian Ministry of Education, University and Research, within the FIRB program (grant number RBFR1280NN). Authors are grateful to Francois Rineau for assistance with mycorrhiza inoculations and to Matteo Garbelotto for helpful comments on the manuscript and for revising the English. The authors wish to thank the Editor and the anonymous Reviewers for insightful suggestions on how to improve the manuscript.

Tunisian desert truffles have been identified on the basis of the internal transcribed spacer (ITS) locus as Tirmania nivea, Terfezia bouderii and T. arenaria. ITS and, for the first time, β-tubulin gene were employed in the phylogenetic analyses of these species to assess whether β-tubulin gene could be a complementary molecular tool for resolution of phylogenetic species. From their comparison, both markers showed to be suitable to distinguish Tirmania nivea from Terfezia bouderii and T. arenaria and, focusing on the T. boudieri complex, to assign the phylogenetic types previously described. In addition to give a first glimpse into the distribution of desert truffles from north to south of Tunisia, we then suggest to use the β-tubulin gene as a complementary marker to ITS. This indication can be useful above all for the phylogenetic reconstruction of the T. boudieri complex.

In major depressive disorder (MDD) patients, life stress events represent a risk factor for a severe, early-onset, treatment-resistant and chronic endophenotype. Treatment-resistant depression (TRD) patients who have experienced traumatic events could benefit from evidence-based trauma-focused psychotherapies. Because this topic has never been investigated, the aim of this pilot trial was to evaluate whether trauma-focused cognitive-behavioural therapy (TF-CBT) and/or eye movement desensitization and reprocessing (EMDR) can help achieve depressive symptom remission in TRD patients. We carried out a single-blind randomized controlled trial with TRD patients and we compared EMDR (N = 12) with TF-CBT (N = 10). Patients received 3 individual sessions per week over a period of 8 weeks. The symptomatological assessments were performed at 4 timepoints: baseline (T0), 4 (T4), 8 (T8) and 12 (T12) weeks. After 24 weeks, a clinical interview was carried out by phone. All TRD patients showed a significant improvement in depressive symptomatology; however, post hoc comparisons showed a significant difference between the two treatment groups, with lower depressive symptom scores in the EMDR than in the TF-CBT group at the follow-up (T12). This effect was partly maintained at 24 weeks. This pilot study suggests that evidence-based trauma-focused psychotherapies, particularly EMDR, can represent effective interventions to treat TRD patients.

Truffles are fungi producing hypogeous fruiting bodies belonging to at least 13 phylogenetically distant orders. The most studied are "true truffles" belonging to the genus Tuber, which is the most economically important group. Truffle fruiting bodies are colonized by bacteria, yeasts, guest filamentous fungi and viruses that, all together, constitute the truffle microbiota. Research on the role of this community has demonstrated that bacteria contribute to truffle aroma. From the ecological point of view, truffle aroma attracts mycophagous animals, which in turn disperse and diffuse truffle spores in the soil, and mediates interactions with microorganisms and plant roots. Truffles have a heterothallic organization, whereby for truffle reproduction it is necessary that strains of opposite mating type meet. Regarding truffle development, the truffle ascocarps use carbon coming from the host plant and not from dead host tissues or soil organic matter as believed so far. In addition to form ectomycorrhizae with a wide diversity of host plants, some truffle species are able to form also arbutoid and orchid mycorrhizas. Knowledge of truffle diversity, traditionally relied on the survey and molecular identification of fruiting bodies, moved over the years towards the survey of mycorrhizas and, recently, on the distribution in soil of the mycelium, with the determination of genets and mating types. The possibility of studying (micro) organisms directly in the field (metagenomics or environmental genomics) and the introduction of high-throughput sequencing techniques (454 pyrosequencing) have given a strong impulse to the study of the microbial communities interacting with truffles and their habitat.

The effects of plant symbionts on host defence responses against pathogens have been extensively documented, but little is known about the impact of pathogens on the symbiosis and if such an impact may differ for nonnative and native pathogens. Here, this issue was addressed in a study of the model system comprising Pinus pinea, its ectomycorrhizal symbiont Tuber borchii, and the nonnative and native pathogens Heterobasidion irregulare and Heterobasidion annosum, respectively. In a 6-month inoculation experiment and using both in planta and gene expression analyses, we tested the hypothesis that H. irregulare has greater effects on the symbiosis than H. annosum. Although the two pathogens induced the same morphological reaction in the plant-symbiont complex, with mycorrhizal density increasing exponentially with pathogen colonization of the host, the number of target genes regulated in T. borchii in plants inoculated with the native pathogen (i.e. 67% of tested genes) was more than twice that in plants inoculated with the nonnative pathogen (i.e. 27% of genes). Although the two fungal pathogens did not differentially affect the amount of ectomycorrhizas, the fungal symbiont perceived their presence differently. The results may suggest that the symbiont has the ability to recognize a self/native and a nonself/nonnative pathogen, probably through host plant-mediated signal transduction.

In this work, we aim at investigating the performance of an Italian Agrammatic speaker with compound words, with major emphasis on the processing of (complex and simple) prepositions inside words, thus aiming at especially evaluating the performance with prepositional compounds of the [NOUN HEAD-PREP-DEPENDANT NOUN; N-P-N] form (coda di cavallo, horse-tail) and exocentric compounds of the [PREP-NOUN; P-N] form (sopracciglio, eyebrow). Bisetto and Scalise (1999) showed that it is realistic to consider N-P-N items as fully productive compound words in Italian, due to the fact that they obey to a set of classic compound-hood tests. Our results demonstrate that SM is selectively impaired in retrieving the prepositions linking the modifying nouns to their head. Our data can trigger interesting interpretations, from a theoretical viewpoint. In particular, complex prepositions (e.g. fuori, outside), which are produced with no significant problems by SM, are likely to be relational nouns and not functional axial parts ( Svenonius, 2006 ) when involved in the formation of Italian P-N compounds. Otherwise, a deficit in retrieving them correctly would be expected (specific agrammatic deficits for axial parts have been detected in Zampieri et al., 2011 ). Moreover, a crucial question is raised: are N-P-N real compounds, since they behave very differently from other compounds in SM performance? Possibly, the same underlying architecture holds both when these items are processed as phrased and when they are processed as “lexicalized syntax” in a constructionist fashion ( Jackendoff, 2002 , Booij, 2005 , Starke, 2009 ). Given the very similar poor performance of SM with both N-P-N compound-like-items and analogous phrases, a unified analysis of this sort is strongly suggested by our study.

Zampieri, Elisa; Franco, Ludovico; Bencini, Giulia, and Meneghello, Francesca(2013), “On the Production of Axial Prepositions: Linking Figure and Ground in Broca`s Aphasia: A Case Study”, Language & Information Society 19. We report the case of a classic agrammatic speaker of Italian in repeating locative Figure/Ground relations via complex prepositions consisting of a locative axial preposition and a simple preposition in light of recent theoretical accounts of prepositions and the existing literature on prepositions in aphasia. The repetition data showed greater rates of omission for simple than axial prepositions, and a trade-off between the inclusion of the Figure and the Axial preposition. Grounds tended to be preserved. These results are unexpected on accounts that simple prepositions are more functional in nature than axial prepositions and argue for an analysis in which axial prepositions can take on a functional grammatical role. To account for the repetition data we propose that the agrammatic processor is unable to fill and retain functional Axial Parts. Hence, it links Figure and Ground through a reduced configuration, mediated by the simple preposition operating as a relational item and not a functional category. The clinical data presented here add to the empirical data set against which to test the validity of finergrained distinctions among prepositions and compare different theoretical views of the status of prepositions.

A major goal in ecology is to understand how interactions among organisms influence ecosystem services. This work compares the effects of two Lepidoptera defoliators, one non-native (Hyphantria cunea) and one native (Lymantria dispar) to Europe, on the colonization of black poplar (the Populus nigra clone “Jean Pourtet”) roots by an arbuscular mycorrhizal (AM) symbiotic fungus (Funneliformis mosseae) in a pot experiment. The effects of defoliation have also been assessed on the expression of fungal and plant genes playing a role during symbiosis. Both control and defoliated poplars have shown a low level of mycorrhization. Additionally, neither the non-native nor the native insect seem to strongly affect the AM colonization, at least at the time of observation (eight days from the end of the defoliation). Concerning the gene expression analysis, our results suggest that defoliation does not influence neither the expression of genes coding for a fungal and a plant phosphate transporter nor that of a gene coding for a fungal ATPase, and that there were no differences between defoliation carried out by the non-native and the native insect.

For some truffle species of the Tuber genus, the symbiotic phase is often associated with the presence of an area of scant vegetation, commonly known as the brûlé, around the host tree. Previous metagenomics studies have identified the microorganisms present inside and outside the brûlé of a Tuber melanosporum truffle-ground, but the molecular mechanisms that operate in this ecological niche remain to be clarified. To elucidate the metabolic pathways present in the brûlé, we conducted a metaproteomics analysis on the soil of a characterized truffle-ground and cross-referenced the resulting proteins with a database we constructed, incorporating the metagenomics data for the organisms previously identified in this soil. The soil inside the brûlé contained a larger number of proteins and, surprisingly, more proteins from plants, compared with the soil outside the brûlé. In addition, Fisher’s Exact Tests detected more biological processes inside the brûlé; these processes were related to responses to multiple types of stress. Thus, although the brûlé has a reduced diversity of plant and microbial species, the organisms in the brûlé show strong metabolic activity. Also, the combination of metagenomics and metaproteomics provides a powerful tool to reveal soil functioning.

International audience; The Tuber melanosporum genome has been analysed with the aim of identifying and characterizing the genes involved in the environmental stress response. A whole genome array (7496 genes/probe) was used to verify the fungal transcriptional profiling upon a cold temperature period (7 days at 4 degrees C). A total of 423 genes resulted to be differentially expressed in a significant manner (>2.5-fold; p-value

Characterization of genomic variation among different microbial species, or different strains of the same species, is a field of significant interest with a wide range of potential applications. We have investigated the genomic variation in mycorrhizal fungal genomes through genomic suppressive subtractive hybridization. The comparison was between phylogenetically distant and close truffle species (Tuber spp.), and between isolates of the ericoid mycorrhizal fungus Oidiodendron maius featuring different degrees of metal tolerance. In the interspecies experiment, almost all the sequences that were identified in the Tuber melanosporum genome and absent in Tuber borchii and Tuber indicum corresponded to transposable elements. In the intraspecies comparison, some specific sequences corresponded to regions coding for enzymes, among them a glutathione synthetase known to be involved in metal tolerance. This approach is a quick and rather inexpensive tool to develop molecular markers for mycorrhizal fungi tracking and barcoding, to identify functional genes and to investigate the genome plasticity, adaptation and evolution.

The huge diversity of fungi may reflect both the heterogeneity of the niches they occupy and the diverse stresses they must cope with. In order to investigate the genetic and functional diversity in the ericoid mycorrhizal fungus Oidiodendron maius subjected to heavy metal stress, we isolated O. maius strains from a serpentine site naturally enriched by heavy metals. Despite the high Cr and Ni soil concentrations, a high level of diversity was found in the serpentine fungal community. The growth of these isolates in the presence of different metal contaminants identified some tolerant strains, suggesting a site-specific adaptation. To investigate within-species gene divergence in stressful environments, we then compared the sequence polymorphism of a neutral (internal transcribed spacer) and a functional (Cu,ZnSOD) gene in O. maius isolates derived from the serpentine site, from a site heavily polluted with industrial wastes and from unpolluted sites. For all isolates tested, the polymorphism was higher in the nucleotide sequence of the functional gene. However, when compared with isolates from the serpentine area, isolates from industrially polluted sites showed a significantly higher polymorphism in the Cu,ZnSOD promoter region, suggesting that environmental stress may influence the rate of mutations in specific regions of the Sod1 locus.

Truffles are hypogeous ectomycorrhizal fungi. They belong to the genus Tuber and are currently considered a hot spot in fungal biology due to their ecological and economic relevance. Among all the species, Tuber magnatum is the most appreciated because of its special taste and aroma. The aim of this work was to set up a protocol to detect T. magnatum in soil and to assess its distribution in a natural truffle-ground. We used the β-tubulin gene as a marker to identify T. magnatum in the soil. This gene allowed us to trace the distribution of the fungus over the entire truffle-ground. Tuber magnatum was found, in one case, 100 m from the productive host plant. This study highlights that T. magnatum mycelium is more widespread than can be inferred from the distribution of truffles and ectomycorrhizas. Interestingly, a new haplotype – never described from fruiting body material – was identified. The specific detection of T. magnatum in the soil will allow to unravel the ecology of this fungus, following its mycelial network. Moreover, this new tool may have practical importance in projects aimed to increase large-scale truffle production, checking for T. magnatum persistence in plantations.

Truffles are hypogeous Ascomycete fungi belonging to the genus Tuber and forming fruiting bodies highly prized for their taste and aroma. The identification of the genus Tuber and its species is important to investigate their ecology and avoid fraud in the food market. As genus-specific primers are not available, the aims of this work were (1) to assess the usefulness of the beta-tubulin gene as a DNA barcoding region for designing Tuber genus-specific primers, (2) to test the primers on a range of fruiting bodies, representing a large part of truffle biodiversity and (3) to check their ecological usefulness, applying them to truffle-ground soil. The new primers designed on the beta-tubulin gene were specific to the Tuber genus in nested PCR. When applied to DNA from soils, they gave a positive signal for 23 of 32 soils. Phylogenetic analysis confirmed that the bands corresponded to Tuber and that at least five Tuber species were present in the truffle-ground. beta-tubulin was found to be a good barcoding region for designing Tuber genus-specific primers, detecting a high Tuber diversity in a natural environment. These primers will be useful for understanding truffle ecology and for practical needs in plantation management.

The effects of biological invasions by non-native species have been widely studied in terms of environmental, economic, and human health impacts. However, little is known about the consequences that non-native plant pathogens may determine on host plant symbionts, such as ectomycorrhizal (ECM) fungi. In this study, interactions between Suillus luteus, an ECM fungus of pine trees, and the allopatrically differentiated fungal pathogens of pines Heterobasidion irregulare and H. annosum were investigated in dual culture by morphological and gene expression analyses. Growth of S. luteus was inhibited by both Heterobasidion species, but based on statistical analysis, growth inhibition was due to the isolate rather than to the species. The expression analysis on genes related to cell wall hydrolytic enzymes and hydrophobins, putatively involved in the fungus-fungus interaction, allowed to identify significantly up- and down-regulated genes both in the symbiont and in the pathogens. Based on the transcript analysis, it was not possible to distinguish the impact of the two pathogenic species on the ECM fungus. The only exception was a S. luteus gene coding for a putative chitinase (SlGH18_8356) that was found to be differentially regulated during interaction with H. irregulare compared to H. annosum.

Objectives Electroconvulsive therapy (ECT) is the most effective therapy for patients with treatment-resistant depression; however, some patients do not respond or relapse in a short time. Electroconvulsive therapy stimulus parameters may be related to the outcome. We carried out a retrospective study review to investigate various ECT parameters in relation to the outcome, clinical variables, and pharmacological treatments. Our aim was to understand which factors could be considered putative seizure quality markers and which are relevant to clinical practice. Methods Two physicians evaluated the seizure length, the postictal suppression index, the wave amplitude, tachycardia, and hemispheric brain wave synchronicity in a double-blind manner for 45 treatment-resistant depression patients receiving ECT. Results The analysis showed a significant association between the outcome and the ECT seizure quality measured by the parameters (P = 9.9 × 10−5). Among patients with poor-quality seizures, 61.5% relapsed after approximately 1 month from the last ECT session. Particularly, there was an association between higher symptomatology decrease and higher quality of hemispheric brain wave synchronicity (P = 5.0 × 10−6), as well as a higher wave amplitude (P = 0.01). Conclusions Our results confirm that ECT seizure quality was strongly correlated with the decrease of depressive symptomatology.

Ectomycorrhizal (ECM) fungi form association with relatively small number of plants that dominate boreal, temperate, Mediterranean, and some subtropical forest ecosystems. These plant species have been able to acquire metabolic capabilities through symbioses with ECM fungi, thus improving their mineral nutrition and growth in several ecological niches. Mycorrhizal fungi can also play several other important ecological roles, including the protection of plants from abiotic and biotic stresses. Several "targeted" metagenomic projects have been carried out, or are now in progress, in order to identify the fungal communities in soil, including ECM fungi, which are present in various habitats (e.g., forest and truffl eground soils, etc.). ECM fungi, which are important both because of their economic value as edible fungi (i.e., truffl es, boletes) and because of their application in reforestation projects, are the subject of this chapter, in which the recent advances in ECM fungal communities are reviewed, focusing mainly on the applicative aspects related to the use of these fungi.

The aim of this study was to investigate the impact of different 4 °C post-harvest storage periods on the quality of the white truffle Tuber magnatum. The expression of selected genes and the profiles of non-volatile metabolites have been analyzed. The up-regulation of genes related to cell wall metabolism and to a putative laccase points to cell wall modifications and browning events during cold storage. Time course RT-qPCR experiments have demonstrated that such transcription events probably depend on the ripening status, since this is delayed in partially ripe fruiting bodies. Changes in the concentrations of linoleate-derived metabolites occur during the first 3 days of considered cold storage, while the other metabolites, such as the amino acids, do not change. Taken together, the results demonstrate that complex molecular events occur in white truffles in the post-harvest period and before they are used as fresh products. © 2014 Springer-Verlag Berlin Heidelberg.

Truffles are hypogeous ectomycorrhizal fungi. Of all the different species, Tuber melanosporum is one of the most popular on the truffle market. The aim of this work was to set up a protocol in order to check the fertility of a T. melanosporum ground. The correlation between its abundance in soil, the presence of mating type genes and productivity was investigated. Soil sampling was conducted in a truffle-ground over two periods of the T. melanosporum life cycle, and under two different host species, to verify whether the time and plant species can affect the quantity of mycelium in the soil. An effective quantitative PCR protocol was set up and employed to the investigated truffle-orchard. We found a statistically significant difference in T. melanosporum abundance between the productive and unproductive soils collected in April. Mating type genes for T. melanosporum were detected under productive and formally productive trees and generally not under unproductive trees even though T. melanosporum was detected. In all the three situations the mating type genes were detected when more than 0.3 ng of T. melanosporum DNA was present. Our results suggest combining these approaches to increase knowledge on the fertility of truffle orchards.

Characterization of genomic variation among different microbial species, or different strains of the same species, is a field of significant interest with a wide range of potential applications. We have investigated the genomic variation in mycorrhizal fungal genomes through genomic suppressive subtractive hybridization. The comparison was between phylogenetically distant and close truffle species (Tuber spp.), and between isolates of the ericoid mycorrhizal fungus Oidiodendron maius featuring different degrees of metal tolerance. In the interspecies experiment, almost all the sequences that were identified in the Tuber melanosporum genome and absent in Tuber borchii and Tuber indicum corresponded to transposable elements. In the intraspecies comparison, some specific sequences corresponded to regions coding for enzymes, among them a glutathione synthetase known to be involved in metal tolerance. This approach is a quick and rather inexpensive tool to develop molecular markers for mycorrhizal fungi tracking and barcoding, to identify functional genes and to investigate the genome plasticity, adaptation and evolution.

Persons with traumatic brain injury (TBI) often show impaired linguistic and/or narrative abilities. The present study aimed to document the features of narrative discourse impairment in a group of adults with TBI. 14 severe TBI non-aphasic speakers (GCS

Letter; International audience; The Périgord black truffle ($Tuber\ melanosporum$ Vittad.) and the Piedmont white truffle dominate today's truffle market. The hypogeous fruiting body of $T.\ melanosporum$ is a gastronomic delicacy produced by an ectomycorrhizal symbiont endemic to calcareous soils in southern Europe. The worldwide demand for this truffle has fuelled intense efforts at cultivation. Identification of processes that condition and trigger fruit body and symbiosis formation, ultimately leading to efficient crop production, will be facilitated by a thorough analysis of truffle genomic traits. In the ectomycorrhizal $Laccaria\ bicolor$, the expansion of gene families may have acted as a 'symbiosis toolbox'. This feature may however reflect evolution of this particular taxon and not a general trait shared by all ectomycorrhizal species. To get a better understanding of the biology and evolution of the ectomycorrhizal symbiosis, we report here the sequence of the haploid genome of $T.\ melanosporum$, which at $\sim$125 megabases is the largest and most complex fungal genome sequenced so far. This expansion results from a proliferation of transposable elements accounting for $\sim$58% of the genome. In contrast, this genome only contains $\sim$7,500 protein-coding genes with very rare multigene families. It lacks large sets of carbohydrate cleaving enzymes, but a few of them involved in degradation of plant cell walls are induced in symbiotic tissues. The latter feature and the upregulation of genes encoding for lipases and multicopper oxidases suggest that $T.\ melanosporum$ degrades its host cell walls during colonization. Symbiosis induces an increased expression of carbohydrate and amino acid transporters in both $L.\ bicolor$ and $T.\ melanosporum$, but the comparison of genomic traits in the two ectomycorrhizal fungi showed that genetic predispositions for symbiosis $-$'the symbiosis toolbox'$-$ evolved along different ways in ascomycetes and basidiomycetes