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10 p.-6 fig., Lentinula is a broadly distributed group of fungi that contains the cultivated shiitake mushroom, L. edodes. We sequenced 24 genomes representing eight described species and several unnamed lineages of Lentinula from 15 countries on four continents. Lentinula comprises four major clades that arose in the Oligocene, three in the Americas and one in Asia–Australasia. To expand sampling of shiitake mushrooms, we assembled 60 genomes of L. edodes from China that were previously published as raw Illumina reads and added them to our dataset. Lentinula edodes sensu lato (s. lat.) contains three lineages that may warrant recognition as species, one including a single isolate from Nepal that is the sister group to the rest of L. edodes s. lat., a second with 20 cultivars and 12 wild isolates from China, Japan, Korea, and the Russian Far East, and a third with 28 wild isolates from China, Thailand, and Vietnam. Two additional lineages in China have arisen by hybridization among the second and third groups. Genes encoding cysteine sulfoxide lyase (lecsl) and γ-glutamyl transpeptidase (leggt), which are implicated in biosynthesis of the organosulfur flavor compound lenthionine, have diversified in Lentinula. Paralogs of both genes that are unique to Lentinula (lecsl 3 and leggt 5b) are coordinately up-regulated in fruiting bodies of L. edodes. The pangenome of L. edodes s. lat. contains 20,308 groups of orthologous genes, but only 6,438 orthogroups (32%) are shared among all strains, whereas 3,444 orthogroups (17%) are found only in wild populations, which should be targeted for conservation., This research was supported by the NSF awards DEB-1456588 to D.H. and DEB-1442113 to A.R., the Department of Energy award 0000238431 to D.H., the GENOBIOREF (BIO2017-86559-R) project of the Spanish Ministry of Science and Innovation (cofinanced by FEDER funds) to F.J.R.-D., S.C., and A.T.M., grant PID2019-103901GB-I00 of the Spanish Ministry of Science and Innovation to P.F., and the NIH award R56AI146096 to A.R. J.L.S. and A.R. were supported by the HHMI and the Burroughs Wellcome Fund. The work (proposal: 10.46936/10.25585/60001048) conducted by the US Department of Energy JGI (https://ror.org/04xm1d337), a DOE Office of Science User Facility, was supported by the Office of Science of the US Department of Energy operated under contract no. DE-AC02-05CH11231.

International audience; Summary The ectomycorrhizal (ECM) symbiosis has independently evolved from diverse types of saprotrophic ancestors. In this study, we seek to identify genomic signatures of the transition to the ECM habit within the hyper-diverse Russulaceae. We present comparative analyses of the genomic architecture and the total and secreted gene repertoires of 18 species across the order Russulales of which 13 are newly sequenced, including a representative of a saprotrophic member of Russulaceae, Gloeopeniophorella convolvens. The genomes of ECM Russulaceae are characterized by a loss of genes for plant cell-wall degrading enzymes (PCWDEs), an expansion of genome size through increased transposable element (TE) content, a reduction in secondary metabolism clusters, and an association of small secreted proteins (SSPs) with TE “nests”, or dense aggregations of TEs. Some PCWDEs have been retained or even expanded, mostly in a species-specific manner. The genome of Gloeopeniophorella convolvens possesses some characteristics of ECM genomes (e.g., loss of some PCWDEs, TE expansion, reduction in secondary metabolism clusters). Functional specialization in ectomycorrhizal decomposition may drive diversification. Accelerated gene evolution predates the evolution of the ECM habit, indicating that changes in genome architecture and gene content may be necessary to prime the evolutionary switch.

Cleanup of contaminated waste sites is a National priority to protect human health and the environment, while restoring land to productive uses. While there are uncertainties with understanding risk to individuals from exposure, the aim of this study was to focus on uncertainties and complexities for ecological systems, complicated by hundreds of species occupying any remediation site which participate in multiple-interacting food webs. The ability to better predict the effectiveness of remediation in fostering future ecosystems might facilitate remedy selection and improve strategic environmental management. This investigation examined (1) uncertainties in ecosystem processes, (2) uncertainties in exposure from contamination before remediation, and (3) uncertainties during remediation. Two Department of Energy sites Hanford Site and Savannah River Site were used as case studies to illustrate how the uncertainties affect eco-receptors. Several types of ecological, physical, and human dimension uncertainties are defined. Ecological uncertainties include temporal, spatial, individual, developmental, and exogenous types. Physical uncertainties are weather-related, watershed variations, slope/aspect, soil/sediment structure and form, unforeseen events, and temporal patterns. Human dimension uncertainties include current land use, future land use, extractive and non-extractive recreation. The effects of remedial strategies varied between the two sites because Hanford is a primarily arid shrub-steppe ecotype, while Savannah River is a wet forest ecotype. Defining the associated ecological sensitivities and uncertainties and providing examples might help policy-makers, managers, planners, and contractors to be aware of issues to consider throughout planning, remediation, and restoration. Adding ecological uncertainty analysis to risk evaluations and remediation planning is analogous to using safety factors in human health risk assessment.

Dry dipterocarp forests are among the most common habitat types in Thailand. Russulaceae are known as common ectomycorrhizal symbionts of Dipterocarpaceae trees in this type of habitat. The present study aims to identify collections of Russula subsection Amoeninae Buyck from dry dipterocarp forests in Thailand. A multi-locus phylogenetic analysis placed Thai Amoeninae collections in two novel lineages, and they are described here as Russula bellissima sp. nov. and R. luteonana sp. nov. The closest identified relatives of both species were sequestrate species suggesting that they may belong to drought-adapted lineages. The analysis of publicly available ITS sequences in R. subsect. Amoeninae did not confirm evidence of any of the new species occurring in other Asian regions, indicating that dry dipterocarp forests might harbor a novel community of ectomycorrhizal fungi. Macromorphological characters are variable and are not totally reliable for distinguishing the new species from other previously described Asian Amoeninae species. Both new species are defined by a combination of differentiated micromorphological characteristics in spore ornamentation, hymenial cystidia and hyphal terminations in the pileipellis. The new Amoeninae species may correspond to some Russula species collected for consumption in Thailand, and the detailed description of the new species can be used for better identification of edible species and food safety in the region.

Since 2007, the quality of Russula descriptions has improved and the use of molecular support for species delimitation and the number of published new species has increased. However, the description style is not consistent and has regional or author-specific patterns. Most recent publications still favour descriptions of spores compared to hymenium and pileipellis elements, and usually only the spore size is provided with statistical support. This study proposes standards for descriptions of the microscopic structure of Russula species (Russulaceae, Agaricomycetes). We present the description template, the template measurements table, the specific terminology and the essential chemical reagents. The proposed standards were tested by mycologists from 11 countries who met at the Russula Microscopy Workshop in Slovakia. Descriptions of 26 species from 9 countries and four continents were prepared, among them R. amarissima, R. castanopsidis, R. seperina and R. subtilis are re-described and 15 species are introduced as new: R. abietiphila, R. amerorecondita, R. aurantioflava, R. echidna, R. flavobrunnescens, R. fluvialis, R. fortunae, R. garyensis, R. gemmata, R. laevis, R. madrensis, R. olivaceohimalayensis, R. purpureogracilis, R. sancti-pauli and R. wielangtae. Seven descriptions for candidate new species are provided without a formal name assignment. Pairwise comparison of species described in this study with available similar descriptions of related species suggests that microscopic characters from all parts of the basidiomata can be equally important for species recognition and they deserve the same treatment including number of measurements and statistics. The majority of recent studies does not recognise differences between the pileus margin and centre, but more than one-third of the species described in this study show distinct differences between the pileus areas, emphasizing the importance to specify the origin of pileipellis observations. This study proved that there is frequently insufficient difference in the ITS barcode between closely related species and that it is necessary to use more genetic markers combined with ecological and geographical data. © 2019, School of Science.

The ectomycorrhizal symbiosis is an essential guild of many forested ecosystems and has a dynamic evolutionary history across kingdom Fungi, having independently evolved from diverse types of saprotrophic ancestors. In this study, we seek to identify genomic features of the transition to the ectomycorrhizal habit within the Russulaceae, one of the most diverse lineages of ectomycorrhizal fungi. We present comparative analyses of the pangenome and gene repertoires of 21 species across the order Russulales, including a closely related saprotrophic member of Russulaceae. The ectomycorrhizal Russulaceae is inferred to have originated around the Cretaceous-Paleogene extinction event (73.6-60.1 million years ago (MY)). The genomes of the ectomycorrhizal Russulaceae are characterized by a loss of genes for plant cell-wall degrading enzymes (PCWDEs), an expansion of genome size through increased transposable element (TE) content, a reduction in secondary metabolism clusters, and an association of genes coding for certain secreted proteins with TE “nests”. The saprotrophic sister group of the ectomycorrhizal Russulaceae, Gloeopeniophorella convolvens, possesses some of these aspects (e.g., loss of some PCWDE and protease orthologs, TE expansion, reduction in secondary metabolism clusters), resulting from an accelerated rate of gene evolution in the shared ancestor of Russulaceae that predates the evolution of the ectomycorrhizal habit. Genomes of Russulaceae possess a high degree of synteny, including a conserved set of terpene secondary metabolite gene clusters. We hypothesize that the evolution of the ectomycorrhizal habit requires premodification of the genome for plant root association followed by an accelerated rate of gene evolution within the secretome for host-defense circumvention and symbiosis establishment.

We describe the first species of Lentinula from Africa, Lentinula madagasikarensis sp. nov. The new taxon, which was collected from central Madagascar, is strikingly similar to L. edodes, the shiitake mushroom. A BLAST search using ITS sequences from L. madagasikarensis as the query retrieves a mix of Lentinula, Gymnopus, Marasmiellus, and other members of Omphalotaceae as the top hits. A 28S phylogeny of the Omphalotaceae confirms placement of L. madagasikarensis within Lentinula. An ITS phylogeny places L. madagasikarensis as the sister group of L. aciculospora, which is a neotropical species. Lentinula madagasikarensis is characterized by robust basidiomata with vinaceous pilei, prominent floccose scales near the pileus margin, florets of sphaeropedunculate cheilocystidia, and subcylindrical basidiospores. This report constitutes a 4 000-mile, trans-oceanic range extension for Lentinula.

Russula subsection Amoeninae is morphologically defined by a dry velvety pileus surface, a complete absence of cystidia with heteromorphous contents in all tissues, and spores without amyloid suprahilar spot. Thirty-four species within subsection Amoeninae have been published worldwide. Although most Russula species in South Korea have been assigned European or North American names, recent molecular studies have shown that Russula species from different continents are not conspecific. Therefore, the present study aims to: 1) define which species of Russula subsection Amoeninae occur on each continent using molecular phylogenetic analyses; 2) revise the taxonomy of Korean Amoeninae. The phylogenetic analyses using the internal transcribed spacer (ITS) and multilocus sequences showed that subsection Amoeninae is monophyletic within subgenus Heterophyllidiae section Heterophyllae. A total of 21 Russula subsection Amoeninae species were confirmed from Asia, Australia, Europe, North America, and Central America, and species from different continents formed separate clades. Three species were recognized from South Korea and were clearly separated from the European and North American species. These species are R. bella, also reported from Japan, a new species described herein, Russula orientipurpurea, and a new species undescribed due to insufficient material.

Societal Impact Statement Research and publication of the planet's remaining plant and fungal species as yet unknown to science is essential if we are to address the United Nations Sustainable Development Goal (SDG) 15 “Life on Land” which includes the protection of terrestrial ecosystems and halting of biodiversity loss. If species are not known to science, they cannot be assessed on the International Union for Conservation of Nature (IUCN) Red List of Threatened Species and so the possibility to protect them from extinction is reduced. Furthermore, until species are known to science they cannot be fully scientifically evaluated for their potential as new foods, medicines, and products which would help address SDGs 1,2,3, and 8. Summary Scientific discovery, including naming new taxa, is important because without a scientific name, a species is invisible to science and the possibilities of researching its ecology, applications and threats, and conserving it, are greatly reduced. We review new scientific discoveries in the plant and fungal kingdoms, based largely on new names of taxa published in 2019 and indexed in the International Plant Names Index and Index Fungorum. Numbers of new species in both kingdoms were similar with 1942 new species of plant published and 1882 species of fungi. However, while >50% of plant species have likely been discovered, >90% of fungi remain unknown. This gulf likely explains the greater number of higher order taxa for fungi published in 2019: three classes, 18 orders, 48 families and 214 genera versus one new family and 87 new genera for plants. We compare the kingdoms in terms of rates of scientific discovery, globally and in different taxonomic groups and geographic areas, and with regard to the use of DNA in discovery. We review species new to science, especially those of interest to humanity as new products, and also by life‐form. We consider where future such discoveries can be expected. We recommend an urgent increase in investment in scientific discovery of plant and fungal species, while they still survive. Priorities include more investment in training taxonomists, in building and equipping collections‐based research centers for them, especially in species‐rich, income‐poor countries where the bulk of species as yet unknown to science are thought to occur.

Numerous lineages of mushroom-forming fungi have been subject to bursts of diversification throughout their evolutionary history, events that can impact our ability to infer well-resolved phylogenies. However, groups that have undergone quick genetic change may have the highest adaptive potential. As the second largest genus of mushroom-forming fungi,Russulaprovides an excellent model for studying hyper-diversification and processes in evolution that drives it. This study focuses on the morphologically defined group –RussulasubsectionRoseinae. Species hypotheses based on morphological differentiation and multi-locus phylogenetic analyses are tested in theRoseinaeusing different applications of the multi-species coalescent model. Based on this combined approach, we recognize fourteen species inRoseinaeincluding the Albida and wholly novel Magnarosea clades. Reconstruction of biogeographic and host association history suggest that parapatric speciation in refugia during glacial cycles of the Pleistocene drove diversification within theRoseinae, which is found to have a Laurasian distribution with an evolutionary origin in the Appalachian Mountains of eastern North America. Finally, we detect jump dispersal at a continental scale that has driven diversification since the most recent glacial cycles.

The DOE Savannah River Site (SRS) is evaluating the potential applicability of the monitored natural attenuation (MNA) process as a contributor to the understanding of the restoration of its unconfined groundwater aquifer known to be contaminated with the chlorinated hydrocarbon compounds trichloroethylene (TCE) and tetrachloroethylene (PCE). This report discusses the results from aqueous desorption experiments on SRS aquifer sediments from two different locations at the SRS (A/M Area; P-Area) with the objective of providing technically defensible TCE/PCE distribution coefficient (Kd) data and data on TCE/PCE reversible and irreversible sorption behavior needed for further MNA evaluation.

Regulatory protocols generally recognize that destructive processes are the most effective mechanisms that support natural attenuation of chlorinated solvents. In many cases, these destructive processes will be biological processes and, for chlorinated compounds, will often be reductive processes that occur under anaerobic conditions. The existing EPA guidance (EPA, 1998) provides a list of parameters that provide indirect evidence of reductive dechlorination processes. In an effort to gather direct evidence of these processes, scientists have identified key microorganisms and are currently developing tools to measure the abundance and activity of these organisms in subsurface systems. Drs. Edwards and Luffler are two recognized leaders in this field. The research described herein continues their development efforts to provide a suite of tools to enable direct measures of biological processes related to the reductive dechlorination of TCE and PCE. This study investigated the strengths and weaknesses of the 16S rRNA gene-based approach to characterizing the natural attenuation capabilities in samples. The results suggested that an approach based solely on 16S rRNA may not provide sufficient information to document the natural attenuation capabilities in a system because it does not distinguish between strains of organisms that have different biodegradation capabilities. The results of the investigationsmore » provided evidence that tools focusing on relevant enzymes for functionally desired characteristics may be useful adjuncts to the 16SrRNA methods.« less

Over the past three decades, much progress has been made in the remediation of chlorinated solvents from the subsurface. Yet these pervasive contaminants continue to present a significant challenge to the U.S. Department of Energy (DOE), other federal agencies, and other public and private organizations. The physical and chemical properties of chlorinated solvents make it difficult to rapidly reach the low concentrations typically set as regulatory limits. These technical challenges often result in high costs and long remediation time frames. In 2003, the DOE through the Office of Environmental Management funded a science-based technical project that uses the U.S. Environmental Protection Agency's technical protocol (EPA, 1998) and directives (EPA, 1999) on Monitored Natural Attenuation (MNA) as the foundation on which to introduce supporting concepts and new scientific developments that will support remediation of chlorinated solvents based on natural attenuation processes. This project supports the direction in which many site owners want to move to complete the remediation of their site(s), that being to complete the active treatment portion of the remedial effort and transition into MNA. The overarching objective of the effort was to examine environmental remedies that are based on natural processes--remedies such as Monitored Natural Attenuation (MNA) ormore » Enhanced Attenuation (EA). The research program did identify several specific opportunities for advances based on: (1) mass balance as the central framework for attenuation based remedies, (2) scientific advancements and achievements during the past ten years, (3) regulatory and policy development and real-world experience using MNA, and (4) exploration of various ideas for integrating attenuation remedies into a systematic set of ''combined remedies'' for contaminated sites. These opportunities are summarized herein and are addressed in more detail in referenced project documents and journal articles, as well as in the technical and regulatory documents being developed within the ITRC. Three topic areas were identified for development during this project. These areas are: mass balance, Enhanced Attenuation (EA), and new characterization and monitoring tools and approaches to support MNA and EA. Each of these topics is documented in stand alone reports, WSRC-STI-2006-00082, WSRC-STI-2006-00083, and WSRC-STI-2006-00084, respectively. In brief, the mass balance efforts are examining methods and tools to allow a site to be evaluated in terms of a system where the inputs and processes within the system are compared to the outputs from the system, as well as understanding what attenuation processes may be occurring and how likely they are to occur within a system. Enhanced Attenuation is a new concept that is a transition step between primary treatments and MNA, when the natural attenuation processes are not sufficient to allow direct transition from the primary treatment to MNA. EA technologies are designed to either boost the level of the natural attenuation processes or decrease the loading of contaminants to the system for a period of time sufficient to allow the remedial goals to be met over the long-term. For characterization and monitoring, a phased approach based on documenting the site specific mass balance was developed. Tools and techniques to support the approach included direct measures of the biological processes and various tools to support cost-effective long-term monitoring of systems where the natural attenuation processes are the main treatment remedies. The effort revealed opportunities for integrating attenuation mechanisms into a systematic set of ''combined remedies'' for contaminated sites.« less

The Department of Energy (DOE) is sponsoring an initiative to facilitate efficient, effective and responsible use of Monitored Natural Attenuation (MNA) and Enhanced Passive Remediation (EPR) for chlorinated solvents. This Office of Environmental Management (EM) ''Alternative Project,'' focuses on providing scientific and policy support for MNA/EPR. A broadly representative working group of scientists supports the project along with partnerships with regulatory organizations such as the Interstate Technology and Regulatory Council and the U.S. Environmental Protection Agency (EPA). The initial product of the technical working group was a summary report that articulated the conceptual approach and central scientific tenants of the project, and that identified a prioritized listing of technical targets for field research. This report documented the process in which: (1) scientific ground rules were developed, (2) lines of inquiry were identified and then critically evaluated, (3) promising applied research topics were highlighted in the various lines of inquiry, and (4) these were discussed and prioritized. The summary report will serve as a resource to guide management and decision-making throughout the period of the subject MNA/EPR Alternative Project. To support and more fully document the information presented in the summary report, we are publishing a series of supplemental documents thatmore » present the full texts from the technical analyses within the various lines of inquiry (see listing). The following report - documenting our evaluation of the state of the science of the characterization and monitoring process and tools-- is one of those supplemental documents.« less

This report was written for multiple audiences. Each reader will likely approach the subject from a different perspective, seeking different information. The team that developed the document consisted of several technical experts on monitored natural attenuation (MNA) and included participants who were integral in the development of US EPA's chlorinated solvent MNA protocol and the subsequent US EPA and Office of Solid Waste and emergency Response (OSWER) policy directive. The primary assignment to the interdisciplinary team was to generate a summary of the state-of-the-art, particularly for chlorinated solvent applications and to identify high-priority opportunities for advancement. A diverse jury of peers reviewed the draft report; the report was edited in response to their comments.