Fungal Secondary Metabolism of Basidiobolus meristosporus: a Novel Natural Product Reservoir

Secondary metabolites (SMs) play an integral role in the life history of most fungal species. Fungal metabolomic studies provide insights into how fungi perform certain ecological functions, how they compete and interact with other organisms, and the breadth of fungal chemodiversity. Fungal SMs have also been utilized for a variety of applications including as pest control agents in agriculture, as food additives, and as pharmaceuticals to treat a variety of ailments. Dikaryotic fungi have been the primary focus of fungal metabolomic studies, resulting in large groups of Kingdom Fungi remaining understudied in terms of SM production. Fungi from atypical environments, such as gut-associated fungi, have been shown to be effective for the targeted discovery of novel SMs. The herptile gut symbiont, Basidiobolus meristosporus, has gained recent attention in phylogenetic, multiomic, and ecological studies, but its secondary metabolism has yet to be studied extensively. Genome annotations and gene predictions for B. meristosporus CBS 931.73 revealed an elevated potential for SM production compared to other closely related fungal species. Additionally, the need for novel pharmaceuticals to address medically important issues, such as the emergence of multi-drug resistant human pathogens, has stoked a resurgence in natural product discovery (NPD) efforts over the past few decades. This resurgence presents an opportunity for the integration of recent technological advances with traditional NPD methodologies to enable more effective approaches for the discovery of novel bioactive products. Genome-directed NPD can be utilized to mitigate the chance of product rediscovery and better target novel natural products (NPs) and identify taxa to act as novel NP reservoirs. Based on gene predictions and the gut-associated ecology of B. meristosporus CBS 931.73, we hypothesized that NPs extracted from B. meristosporus tissue would exhibit biological activities detectable using traditional NPD screening techniques. The efficacy of B. meristosporus CBS 931.73 as a novel NP reservoir was assessed using an assortment of bottom-up and top-down techniques including genome-directed selection of target biological activities, manipulation of culture conditions, bioactivity-guided fractionation, and computational workflows designed to analyze SM extract profiles and identify bioactive constituents. Our results demonstrate a reproducible differentiation of SM production as a response to controlled variation of culturing conditions for clonal cultures grown in laboratory conditions. SM extracts from cultures grown in certain media conditions, particularly in yeast peptone soluble starch (YPSS) broth, were found to selectively inhibit gram-positive bacteria, including drug-resistant strains of Staphylococcus aureus and Enterococcus faecium. Sixteen unique features with antibacterial activity were identified using computational analyses of LC-MS/MS and bioactivity data. Consistent with SM core gene predictions, fourteen of the putatively active compounds were identified as cyclic peptides. While further isolation and characterization of these compounds is necessary, our findings demonstrate the potential of B. meristosporus as a novel NP reservoir and the efficacy of a hybrid approach to NPD utilizing genome-based direction, traditional bioactivity screening methods, and computational workflows for the assessment of novel NP systems

Phylogenomic analyses of non-Dikarya fungi supports horizontal gene transfer driving diversification of secondary metabolism in the amphibian gastrointestinal symbiont, Basidiobolus

Research into secondary metabolism (SM) production by fungi has resulted in the discovery of diverse, biologically active compounds with significant medicinal applications. However, the fungi rich in SM production are taxonomically restricted to Dikarya, two phyla of Kingdom Fungi, Ascomycota and Basidiomycota. Here, we explore the potential for SM production in Mucoromycota and Zoopagomycota, two phyla of nonflagellated fungi that are not members of Dikarya, by predicting and identifying core genes and gene clusters involved in SM. The majority of non-Dikarya have few genes and gene clusters involved in SM production except for the amphibian gut symbionts in the genus Basidiobolus . Basidiobolus genomes exhibit an enrichment of SM genes involved in siderophore, surfactin-like, and terpene cyclase production, all these with evidence of constitutive gene expression. Gene expression and chemical assays confirm that Basidiobolus has significant siderophore activity. The expansion of SMs in Basidiobolus are partially due to horizontal gene transfer from bacteria, likely as a consequence of its ecology as an amphibian gut endosymbiont

A connectome and analysis of the adult Drosophila central brain.

The neural circuits responsible for animal behavior remain largely unknown. We summarize new methods and present the circuitry of a large fraction of the brain of the fruit fly Drosophila melanogaster. Improved methods include new procedures to prepare, image, align, segment, find synapses in, and proofread such large data sets. We define cell types, refine computational compartments, and provide an exhaustive atlas of cell examples and types, many of them novel. We provide detailed circuits consisting of neurons and their chemical synapses for most of the central brain. We make the data public and simplify access, reducing the effort needed to answer circuit questions, and provide procedures linking the neurons defined by our analysis with genetic reagents. Biologically, we examine distributions of connection strengths, neural motifs on different scales, electrical consequences of compartmentalization, and evidence that maximizing packing density is an important criterion in the evolution of the fly's brain

Phylogenomic Analyses of Non-Dikarya Fungi Supports Horizontal Gene Transfer Driving Diversification of Secondary Metabolism in the Amphibian Gastrointestinal Symbiont, Basidiobolus

Research into secondary metabolism (SM) production by fungi has resulted in the discovery of diverse, biologically active compounds with significant medicinal applications. The fungi rich in SM production are taxonomically concentrated in the subkingdom Dikarya, which comprises the phyla Ascomycota and Basidiomycota. Here, we explore the potential for SM production in Mucoromycota and Zoopagomycota, two phyla of nonflagellated fungi that are not members of Dikarya, by predicting and identifying core genes and gene clusters involved in SM. The majority of non-Dikarya have few genes and gene clusters involved in SM production except for the amphibian gut symbionts in the genus Basidiobolus. Basidiobolus genomes exhibit an enrichment of SM genes involved in siderophore, surfactin-like, and terpene cyclase production, all these with evidence of constitutive gene expression. Gene expression and chemical assays also confirm that Basidiobolus has significant siderophore activity. The expansion of SMs in Basidiobolus are partially due to horizontal gene transfer from bacteria, likely as a consequence of its ecology as an amphibian gut endosymbiont

Phylogenomic Analyses of Non-Dikarya Fungi Supports Horizontal Gene Transfer Driving Diversification of Secondary Metabolism in the Amphibian Gastrointestinal Symbiont, Basidiobolus.

Research into secondary metabolism (SM) production by fungi has resulted in the discovery of diverse, biologically active compounds with significant medicinal applications. The fungi rich in SM production are taxonomically concentrated in the subkingdom Dikarya, which comprises the phyla Ascomycota and Basidiomycota. Here, we explore the potential for SM production in Mucoromycota and Zoopagomycota, two phyla of nonflagellated fungi that are not members of Dikarya, by predicting and identifying core genes and gene clusters involved in SM. The majority of non-Dikarya have few genes and gene clusters involved in SM production except for the amphibian gut symbionts in the genus Basidiobolus Basidiobolus genomes exhibit an enrichment of SM genes involved in siderophore, surfactin-like, and terpene cyclase production, all these with evidence of constitutive gene expression. Gene expression and chemical assays also confirm that Basidiobolus has significant siderophore activity. The expansion of SMs in Basidiobolus are partially due to horizontal gene transfer from bacteria, likely as a consequence of its ecology as an amphibian gut endosymbiont

A connectome and analysis of the adult Drosophila central brain.

The neural circuits responsible for animal behavior remain largely unknown. We summarize new methods and present the circuitry of a large fraction of the brain of the fruit fly Drosophila melanogaster. Improved methods include new procedures to prepare, image, align, segment, find synapses in, and proofread such large data sets. We define cell types, refine computational compartments, and provide an exhaustive atlas of cell examples and types, many of them novel. We provide detailed circuits consisting of neurons and their chemical synapses for most of the central brain. We make the data public and simplify access, reducing the effort needed to answer circuit questions, and provide procedures linking the neurons defined by our analysis with genetic reagents. Biologically, we examine distributions of connection strengths, neural motifs on different scales, electrical consequences of compartmentalization, and evidence that maximizing packing density is an important criterion in the evolution of the fly's brain

A connectome and analysis of the adult Drosophila central brain

The neural circuits responsible for animal behavior remain largely unknown. We summarize new methods and present the circuitry of a large fraction of the brain of the fruit fly Drosophila melanogaster. Improved methods include new procedures to prepare, image, align, segment, find synapses in, and proofread such large data sets. We define cell types, refine computational compartments, and provide an exhaustive atlas of cell examples and types, many of them novel. We provide detailed circuits consisting of neurons and their chemical synapses for most of the central brain. We make the data public and simplify access, reducing the effort needed to answer circuit questions, and provide procedures linking the neurons defined by our analysis with genetic reagents. Biologically, we examine distributions of connection strengths, neural motifs on different scales, electrical consequences of compartmentalization, and evidence that maximizing packing density is an important criterion in the evolution of the fly’s brain