The cryptic lifestyle of most fungi necessitates molecular identification of the guild in environmental studies.Over the past decades, rapid development and affordability of molecular tools have tremendously improved insights of the fungal diversity in all ecosystems and habitats. Yet, in spite of the progress of molecular methods, knowledge about functional properties of the fungal taxa is vague and interpretation of environmental studies in an ecologically meaningful manner remains challenging. In order to facilitate functional assignments and ecological interpretation of environmental studies we introduce a user friendly traits and character database FungalTraits operating at genus and species hypothesis levels. Combining the information from previous efforts such as FUNGuild and Fun Fun together with involvement of expert knowledge, we reannotated 10210 and 151 fungal and Stramenopila genera, respectively. This resulted in a stand-alone spreadsheet dataset covering 17 lifestyle related traits of fungal and Stramenopila genera, designed for rapid functional assignments of environmental studies. In order to assign the trait states to fungal species hypotheses, the scientific community of experts manually categorised and assigned available trait information to 697413 fungal ITS sequences. On the basis of those sequences we were able to summarise trait and host information into 92623 fungal species hypotheses at 1% dissimilarity threshold.

To identify hypothesized missing components of the synaptic G␣ o -G␣ q signaling network, which tightly regulates neurotransmitter release, we undertook two large forward genetic screens in the model organism C. elegans and focused first on mutations that strongly rescue the paralysis of ric-8(md303) reductionof-function mutants, previously shown to be defective in G␣ q pathway activation. Through high-resolution mapping followed by sequence analysis, we show that these mutations affect four genes. Two activate the G␣ q pathway through gain-of-function mutations in G␣ q ; however, all of the remaining mutations activate components of the G␣ s pathway, including G␣ s , adenylyl cyclase, and protein kinase A. Pharmacological assays suggest that the G␣ s pathway-activating mutations increase steady-state neurotransmitter release, and the strongly impaired neurotransmitter release of ric-8(md303) mutants is rescued to greater than wild-type levels by the strongest G␣ s pathway activating mutations. Using transgene induction studies, we show that activating the G␣ s pathway in adult animals rapidly induces hyperactive locomotion and rapidly rescues the paralysis of the ric-8 mutant. Using cell-specific promoters we show that neuronal, but not muscle, G␣ s pathway activation is sufficient to rescue ric-8(md303)'s paralysis. Our results appear to link RIC-8 (synembryn) and a third major G␣ pathway, the G␣ s pathway, with the previously discovered G␣ o and G␣ q pathways of the synaptic signaling network.

We used gain-of-function and null synaptic signaling network mutants to investigate the relationship of the G␣ q and G␣ s pathways to synaptic vesicle priming and to each other. Genetic epistasis studies using G␣ q gain-of-function and null mutations, along with a mutation that blocks synaptic vesicle priming and the synaptic vesicle priming stimulator phorbol ester, suggest that the G␣ q pathway generates the core, obligatory signals for synaptic vesicle priming. In contrast, the G␣ s pathway is not required for the core priming function, because steady-state levels of neurotransmitter release are not significantly altered in animals lacking a neuronal G␣ s pathway, even though these animals are strongly paralyzed as a result of functional (nondevelopmental) defects. However, our genetic analysis indicates that these two functionally distinct pathways converge and that they do so downstream of DAG production. Further linking the two pathways, our epistasis analysis of a ric-8 null mutant suggests that RIC-8 (a receptor-independent G␣ guanine nucleotide exchange factor) is required to maintain both the G␣ q vesicle priming pathway and the neuronal G␣ s pathway in a functional state. We propose that the neuronal G␣ s pathway transduces critical positional information onto the core G␣ q pathway to stabilize the priming of selected synapses that are optimal for locomotion. O NE proposed mechanism for establishing and omi and Kidokoro 2000; Waters and Smith 2000). These electrophysiological studies thus show that both modifying behaviors or memories involves presynaptic changes in synaptic strength, and regulating the pathways can positively affect priming, but they do not reveal how the two pathways interact and why they both amount of synaptic-vesicle-mediated neurotransmitter release at synapses is thought to be critical for changing exist. Highlighting this point, two recent studies showed that the G␣ s pathway is not even required for synaptic presynaptic strength (Kandel and Pittenger 1999; Lin vesicle priming, because evoked release remains norand Scheller 2000; Sudhof 2000). Electrophysiologimal, or even increases, when G␣ s null synapses are subcal studies have defined a "readily releasable," or "primed," jected to low frequency stimulation (Hou et al. 2003; pool of synaptic vesicles, the size of which determines Renden and Broadie 2003). release probability and, therefore, presynaptic strength Providing important insights and yet further adding (Sudhof 2000). The term "primed" can be applied eito the puzzlement about the relative roles of the G␣ q ther to an individual synaptic vesicle or to a synapse and G␣ s pathways, recent research suggests that both (i.e., a primed synapse is one containing a relatively large pathways, directly or indirectly, affect the synaptic abunpool of primed synaptic vesicles). A major challenge is dance of UNC-13, a large, conserved protein that binds to understand the layout and logic of the network of diacylglycerol (DAG) (Maruyama and Brenner 1991), signaling pathway...

Environmental DNA surveys reveal that most fungal diversity represents uncultured species. We sequenced the genomes of eight uncultured species across the fungal tree of life using a new single-cell genomics pipeline. We show that, despite a large variation in genome and gene space recovery from each single amplified genome (SAG), ≥90% can be recovered by combining multiple SAGs. SAGs provide robust placement for early-diverging lineages and infer a diploid ancestor of fungi. Early-diverging fungi share metabolic deficiencies and show unique gene expansions correlated with parasitism and unculturability. Single-cell genomics holds great promise in exploring fungal diversity, life cycles and metabolic potential.

Improved sequencing technologies have profoundly altered global views of fungal diversity and evolution. High throughput sequencing methods are critical for studying fungi due to the cryptic, symbiotic nature of many species, particularly those that are difficult to culture. However, the low coverage genome sequencing (LCGS) approach to phylogenomic inference has not been widely applied to fungi. Here we analyzed 171 Kickxellomycotina fungi using LCGS methods to obtain hundreds of marker genes for robust phylogenomic reconstruction. Additionally, we mined our LCGS data for a set of nine rDNA and protein coding genes to enable analyses across species for which no LCGS data were obtained. The main goals of this study were to: 1) evaluate the quality and utility of LCGS data for both phylogenetic reconstruction and functional annotation, 2) test relationships among clades of Kickxellomycotina, and 3) perform comparative functional analyses between clades to gain insight into putative trophic modes. In opposition to previous studies, our nine-gene analyses support two clades of arthropod gut dwelling species and suggest a possible single evolutionary event leading to this symbiotic lifestyle. Furthermore, we resolve the mycoparasitic Dimargaritales as the earliest diverging clade in the subphylum and find four major clades of Coemansia species. Finally, functional analyses illustrate clear variation in predicted carbohydrate active enzymes and secondary metabolites (SM) based on ecology, i.e., biotroph vs. saprotroph. Saprotrophic Kickxellales broadly lack many known pectinase families compared to saprotrophic Mucoromycota and are depauperate for SM but are enriched for chitinases compared to biotrophic taxa in Zoopagomycota.