Stajich LAMG12 - Indoor Fungi

Fungi in the Built Environment Jason Stajich University of California, Riversidehttp://fungidb.org @hyphaltip @fungalgenomes @fungidb http://lab.stajich.org

Fungal diversity of forms, functions, ecosystems Cryptococcus neoformans X. Lin Coprinopsis cinerea Ellison & Stajich Aspergillus niger. N Read Glomus sp. Univ Sydney Rozella allomycis. James et al Puccinia graminis J. F. Hennen Laccaria bicolor Martin et al. Neurospora crassa. Hickey & Reed Phycomyces blakesleansus T. Ootaki Batrachochytrium dendrobatidis J. Longcore Ustilago maydis Kai Hirdes Amanita phalloides. M Wood Xanthoria elegans. Botany POtD Rhizopus stolonifera. Blastocladiella simplex Stajich & Taylor

Plantae Amoebozoa Choanozoa Metazoa Microsporidia Fungi Rozella Chytridiomycota BlastocladiomycotaMulticellular with Mucoromycotinadifferentiated tissues Entomophthoromycotina Zoopagomycotina Loss of flagellum Kickxellomycotina Glomeromycota Mitotic sporangia Pucciniomycotina Basidiomycota to mitotic conidia Ustilaginomycotina Regular septa Agaricomycotina Taphrinomycotina AscomycotaMeiotic sporangia to Saccharomycotinaexternal meiospores Pezizomycotina 1500 1000 500 0 Millions of years Stajich et al. Current Biol 2009

Fungi interact with many organisms 10.3389/fpls.2011.00100 Betsy Arnold doi: 10.3389/fpls.2011.00100Endophytes Mycorrhiza doi: 10.1016/j.pbi.2009.05.007, F. Martin

Organisms interacting with Fungi - fungi as the host REPORTS to the Midwest Regional Center of Excellence for was supported by the NIH Institutional NRSA T32 SOM Text Biodefense and Emerging Infectious Disease Research GM07067 to the Washington University School of Figs. S1 to S4 (MRCE) and by NIH grant AI53298. The DDRCC is Medicine. Tables S1 and S2 supported by NIH grant DK52574. W.W.L. was supported References Supporting Online Material Plant + Fungus + Mycovirus by the Clinical/Translational Fellowship Program of the MRCE, the W.M. Keck Foundation, and the NIH National www.sciencemag.org/cgi/content/full/315/5811/509/DC1 6 November 2006; accepted 14 December 2006 Research Service Award (NRSA) F32 AI069688-01. P.A.P. Materials and Methods 10.1126/science.1137195 A Virus in a Fungus in a Plant: S2). The 2.2-kb fragment (RNA 1) is involved in virus replication, as both of its ORFs are similar to viral replicases. The first, ORF1a, has 29% Three-Way Symbiosis Required for amino acid sequence identity with a putative RNA-dependent RNA polymerase (RdRp) from Thermal Tolerance the rabbit hemorrhagic disease virus. The amino acid sequence of the second, ORF1b, has 33% identity with the RdRp of a virus of the fungal Luis M. Márquez,1 Regina S. Redman,2,3 Russell J. Rodriguez,2,4 Marilyn J. Roossinck1* pathogen Discula destructiva. These two ORFs overlap and could be expressed as a single DOI: 10.1126/science.1136237 Downloaded from www.sciencemag.org on September 18, 2012 A mutualistic association between a fungal endophyte and a tropical panic grass allows both protein by frameshifting, a common expression organisms to grow at high soil temperatures. We characterized a virus from this fungus that is strategy of viral replicases. The two ORFs of involved in the mutualistic interaction. Fungal isolates cured of the virus are unable to confer RNA 2 have no similarity to any protein with heat tolerance, but heat tolerance is restored after the virus is reintroduced. The virus-infected known function. As in most dsRNA mycovi- fungus confers heat tolerance not only to its native monocot host but also to a eudicot host, ruses, the 5′ ends (21 bp) of both RNAs are which suggests that the underlying mechanism involves pathways conserved between these two conserved. Virus particles purified from C. groups of plants. protuberata are similar to those of other fungal viruses: spherical and ~27 nm in diameter (fig. E ndophytic fungi commonly grow within known mutualistic endophyte, Epichloë festucae, S3). This virus is transmitted vertically in the plant tissues and can be mutualistic in but no phenotype has been associated with this conidiospores. We propose naming this virus some cases, as they allow plant adaptation virus (9). Curvularia thermal tolerance virus (CThTV) toAPPLIED AND ENVIRONMENTAL MICROBIOLOGY, June 2010, p. 4063–4075 Vol. 76, No. 12 to extreme environments (1). A plant-fungal Fungal virus genomes are commonly com- reflect its host of origin and its phenotype.0099-2240/10/$12.00 doi:10.1128/AEM.02928-09 symbiosis between a tropical panic grass from posed of double-stranded RNA (dsRNA) (10). The ability of the fungus to confer heatCopyright © 2010, American Society for Microbiology. All Rights Reserved. geothermal soils, Dichanthelium lanuginosum, Large molecules of dsRNA do not normally tolerance to its host plant is related to the and the fungus Curvularia protuberata allows occur in fungal cells and, therefore, their presence presence of CThTV. Wild-type isolates of C. both organisms to grow at high soil temperatures is a sign of a viral infection (9). Using a protocol protuberata contained the virus in high titers, as in Yellowstone National Park (YNP) (2). Field for nucleic acid extraction with enrichment for evidenced by their high concentration of dsRNA Diverse Bacteria Inhabit Living Hyphae of Phylogenetically and laboratory experiments have shown that when root zones are heated up to 65°C, non- dsRNA (11), we detected the presence of a virus in C. protuberata. The dsRNA banding pattern (~2 mg/g of lyophilized mycelium). However, an isolate obtained from sectoring (change in Diverse Fungal Endophytesᰔ† symbiotic plants either become shriveled and consists of two segments of about 2.2 and 1.8 kb. morphology) of a wild-type colony contained a chlorotic or simply die, whereas symbiotic plants A smaller segment, less than 1 kb in length, was very low titer of the virus, as indicated by a low tolerate and survive the heat regime. When variable in presence and size in the isolates concentration of dsRNA (~0.02 mg/g of lyophi- grown separately, neither the fungus nor the plant analyzed and, later, was confirmed to be a sub- lized mycelium). These two isolates were iden- Michele T. Hoffman and A. Elizabeth Arnold* alone is able to grow at temperatures above 38°C, genomic element, most likely a defective RNA tical by simple sequence repeat (SSR) analysis but symbiotically, they are able to tolerate ele- (fig. S1 and Fig. 1, A and B). Using tagged with two single-primer polymerase chain reac- Downloaded from http:// Division of Plant Pathology and Microbiology, School of Plant Sciences, 1140 E. South Campus Drive, vated temperatures. In the absence of heat stress, random hexamer primers, we transcribed the tion (PCR) reactions and by sequence analysis of symbiotic plants have enhanced growth rate virus with reverse transcriptase (RT), followed by the rDNA ITS1-5.8S-ITS2 region (figs. S4 and University of Arizona, Tucson, Arizona 85721 compared with nonsymbiotic plants and also amplification and cloning. Sequence analysis S5). Desiccation and freezing-thawing cycles are show significant drought tolerance (3). revealed that each of the two RNA segments known to disrupt virus particles (12); thus, my- Received 3 December 2009/Accepted 20 April 2010 Fungal viruses or mycoviruses can modulate contains two open reading frames (ORFs) (fig. celium of the isolate obtained by sectoring was plant-fungal symbioses. The best known exam- ple of this is the hypovirus that attenuates the Both the establishment and outcomes of plant-fungus symbioses can be inﬂuenced by abiotic factors, the virulence (hypovirulence) of the chestnut blight Fig. 1. Presence or absence of CThTV in different strains of C. interplay of fungal and plant genotypes, and additional microbes associated with fungal mycelia. Recently fungus, Cryphonectria parasitica (4). Virus regu- protuberata, detected by ethid- lation of hypovirulence has been demonstrated bacterial endosymbionts were documented in soilborne Glomeromycota and Mucoromycotina and in at least Domestication: Ant farmed fungi experimentally in several other pathogenic fungi ium bromide staining (A), Northern blot using RNA 1 (B) one species each of mycorrhizal Basidiomycota and Ascomycota. Here we show for the ﬁrst time that phylo- (5–8). However, the effect of mycoviruses on and RNA 2 (C) transcripts of mutualistic fungal endophytes is unknown. There genetically diverse endohyphal bacteria occur in living hyphae of diverse foliar endophytes, including repre- is only one report of a mycovirus from the well- the virus as probes, and RT- PCR using primers specific for sentatives of four classes of Ascomycota. We examined 414 isolates of endophytic fungi, isolated from photo-

Estimates of the number of species of Fungi Mycol. Res. 9S (6): 641--655 (1991) Printed in Great Britain 641 Presidential address 1990 1.5 Million based on fungus to The fungal dimension of biodiversity: magnitude, significance, plant ratio of 6:1 and conservation D. L. HAWKSWORTH International Mycological Institute, Kew, Surrey TW9 3AF, UK American Journal of Botany 98(3): 426–438. 2011. Don’t forget the endophytes... Fungi, members of the kingdoms Chromista, Fungi S.str. and Protozoa studied by mycologists, have received scant consideration in discussions on biodiversity. The number of known species is about 69000, but that in the world is conservatively estimated at THE FUNGI: 1, 2, 3 … 5.1 MILLION SPECIES?1 and the soil... 15 million; six-times higher than hitherto suggested. The new world estimate is primarily based on vascular plant:fungus ratios in different regions. It is considered conservative as: (1) it is based on the lower estimates of world vascular plants; (2) no separate Meredith Blackwell2 provision is made for the vast numbers of insects now suggested to exist; (3) ratios are based on areas still not fully known mycologically; and (4) no allowance is made for higher ratios in tropical and polar regions. Evidence that numerous new species Department of Biological Sciences; Louisiana State University; Baton Rouge, Louisiana 70803 USA remain to be found is presented. This realization has major implications for systematic manpower, resources, and classification. Fungi DOI:10.3732/ajb.1000298 • Premise of the study: Fungi are major decomposers in certain ecosystems and essential associates of many organisms. They have and continue to playa vital role in the evolution of terrestrial life (especially through mutualisms), ecosystem function and the provide enzymes and drugs and serve as experimental organisms. In 1991, a landmark paper estimated that there are 1.5 million maintenance of biodiversity, human progress, and the operation of Gaia. Conservation in situ and ex situ are complementary, andon the Earth. Because only 70 000 fungi had been described at that time, the estimate has been the impetus to search for fungi the significance of culture collections is stressed. International collaboration is required to develop a world inventory, quantify functional unknown fungi. Fungal habitats include soil, water, and organisms that may harbor large numbers of understudied previously roles, and for effective conservation. fungi, estimated to outnumber plants by at least 6 to 1. More recent estimates based on high-throughput sequencing methods Upwards of 6M species - Lee Taylor (pers comm) suggest that as many as 5.1 million fungal species exist. • Methods: Technological advances make it possible to apply molecular methods to develop a stable classiﬁcation and to dis- cover and identify fungal taxa. Biodiversity, the extent of biological variation on Earth, has species, or populations. Knowledge of all of theseKey pertinent • is results: Molecular methods have dramatically increased our knowledge of Fungi in less than 20 years, revealing a mono-“Thus, the Fungi is likely equaled only by the Insecta with respect to eukaryote species richness.” come to the fore as a key issue in science and politics for the 1990s. First used as BioDiversity in the title of a scientific to a thorough appreciation of the fungal dimension, butkingdom and increased diversity among early-diverging lineages. Mycologists are making signiﬁcant advances in phyletic here I will centre on species biodiversity; that is basal to discussions but many fungi remain to be discovered. species discovery, • Conclusions: Fungi are essential to the survival of many groups of organisms with which they form associations. They also

Fungal genome sequencing 400+ genomes of Fungihttp://www.diark.org/diark/statistics

http://1000.fungalgenomes.org

Addressing the phylogenetic diversity: 1000 Fungal genomes project !"#$%&%()*+#+,- !"#$%&%()*+#+,- .%#$/+%()*+#+,- .%#$/+%()*+#+,- 01"%2%()*+#+,- 01"%2%()*+#+,- D+%E8%,,%()*+#+,- D+%E8%,,%()*+#+,- F+GG%()*%2&4- 3&*+"#4+-,+/,- F+GG%()*%2&4- 3&*+"#4+-,+/,- 547%187+&%()*+#+,- 547%187+&%()*+#+,- 5+*4&%"%()*+#+,- 5+*4&%"%()*+#+,- 5+%2%()*+#+,- 5+%2%()*+#+,- 5*$&%()*+#+,- 5*$&%()*+#+,- 6"78%()*+#+,- 6"78%()*+#+,- F+GG%()*+#+,- F+GG%()*+#+,- H%"/4"%()*+#+,- H%"/4"%()*+#+,- H4**$4"%()*%2&4- H4**$4"%()*+#+,- H4**$4"%()*%2&4- H4**$4"%()*+#+,- I+%8+*#%()*+#+,- I+%8+*#%()*+#+,- J4K$"&%()*%2&4- F&+1(%*),2/%()*+#+,- J4K$"&%()*%2&4- F&+1(%*),2/%()*+#+,- H*$G%,4**$4"%()*+#+,- H*$G%,4**$4"%()*+#+,- J4K$"&%()*+#+,- J4K$"&%()*+#+,- M,284E&%()*%2&4- 0L%74,/%()*+#+,- M,284E&%()*%2&4- 0L%74,/%()*+#+,- M,284E&%()*+#+,- M,284E&%()*+#+,- !E4"*%,287%()*+#+,- !E4"*%,287%()*+#+,- !#"4*2+88%()*+#+,- !#"4*2+88%()*+#+,- N84,,*18%()*+#+,- N84,,*18%()*+#+,- F1**&%()*%2&4- N")K#%()*%*%84*%()*+#+,- F1**&%()*%2&4- N")K#%()*%*%84*%()*+#+,- N),#%74,/%()*+#+,- N),#%74,/%()*+#+,- O*"%7%#")%()*+#+,- O*"%7%#")%()*+#+,- OL%()*+#+,- OL%()*+#+,- F1**&%()*+#+,- F1**&%()*+#+,- !E4"*%()*+#+,- !E4"*%()*%2&4- !E4"*%()*+#+,- .4*")()*+#+,- !E4"*%()*%2&4- .4*")()*+#+,- J"+(+88%()*+#+,- J"+(+88%()*+#+,- D8%(+"%()*+#+,- 0&#%(%K$#$%"%()*%2&4- D8%(+"%()*+#+,- P*Q+88%()*%2&4- 0&#%(%K$#$%"%()*%2&4- O%"2+"+88%()*%2&4- P*Q+88%()*%2&4- 04"8)-.T+"E&E- O1*%"%()*%2&4- O%"2+"+88%()*%2&4- R%%K4E%()*%2&4- 04"8)-.T+"E&E- O1*%"%()*%2&4- 5&+4E+,- R%%K4E%()*%2&4- I+%*488(4,2E%()*+#+,- S84,#%*84/%()*+#+,- 5&+4E+,- I+%*488(4,2E%()*+#+,- N$)#"/%()*+#+,- S84,#%*84/%()*+#+,- O%&%78+K$4"/%()*+#+,- N$)#"/%()*+#+,- O%&%78+K$4"/%()*+#+,- 9- <9- >9- @9- B9- ;99- ;<9- 9:- ;9:- <9:- =9:- >9:- ?9:- @9:- A9:- B9:- C9:- ;99:-Blue = completed or in progress, Red= proposed for Tier One sampling,Green = remaining unsampled families Numbers or Percent of Families in each clade and their current or proposed genome sampling

FungiDB Strategy queries Genome Browser Data-mining SyntenyFunctional PhylogenomicGenomics proﬁles Data Annotation & Function Gene function curation tool

• Eurotiomycetes; Ascomycota ◦ Aspergillus clavatus ◦ Aspergillus flavus ◦ Aspergillus fumigatus strain Af293 ◦ Aspergillus nidulans strain A4 FungiDB 2.0 ◦ Aspergillus niger ◦ Aspergillus terreus ◦ Coccidoidies immitis strain RS ◦ Coccidoidies immitis strain H538.4 • Sordariomycetes; Ascomyocta ◦ Fusarium oxysporum f. sp. lycopersici• 31 genomes - 25 Fungi and 6 Oomycetes ◦ Fusarium graminearum ◦ Gibberella moniliformis (Fusarium verticillioides) ◦ Magnaporthea oryzae• RNA-Seq for 6 species, Microarray for 2 sp, KEGG, EC, ◦ Neurospora crassa strain OR74A ◦ Neurospora tetrasperma and GO annotations for several species ◦ Neurospora discreta ◦ Sordaria macrospora • Taphrinomycotina; Ascomyocta ◦ Schizosaccharomyces pombe• Ortholog tables with OrthoMCL • Saccharomycotina; Ascomyocta ◦ Candida albicans ◦ Saccharomyces cerevisiae• Gene function predictions from InterPro • Basidiomycota ◦ Cryptococcus neoformans var. grubii) strain H99 ◦ Cryptococcus neoformans var. neoformans) strain JEC21 ◦ Cryptococcus neoformans var. neoformans) strain B3501 ◦ Cryptococcus gattii strain WM276 ◦ Cryptococcus gattii strain R265 ◦ Tremella mesenterica • Oomycetes; Stramenopiles ◦ Phytophthora capsici ◦ Phytophthora infestans ◦ Phytophthora ramorum ◦ Phytophthora sojae ◦ Pythium ultimatum ◦ Hyaloperonospora arabidopsidis

Combining queries• Results from one query combined with a second one.• Can be intersection, union, or left or right overlaps

Microbial Ecology of Indoor Fungi• Sloan Foundation initiative to provide a data coordination center for indoor microbiome data• In collaboration with Rob Knight (QIIME), Mitch Sogin (VAMPS), Folker Meyer (MG-RAST)• Fungi - names and taxonomy in ﬂux• Marker Genes and data collection approaches• A sample indoor environment dataset analysis

Fungal Taxonomy and naming undergoing a revolution One fungus, one name IMA FuNgus · voluMe 2 · No 1: 105–112 The Amsterdam Declaration on Fungal Nomenclature A RT I C L E 1 3 3 Taylor 6 *, Özlem Abaci7 , Ahmet Asan , Feng-Yan Bai10 6 , Dominik Begerow11, Derya Berikten , Teun Boekhout 13 , Treena Burgess , Walter Buzina 16 17 , Ulrike Damm , Irina Druzhinina , Ursula Eberhardt , 10 , 30 31 , Ahmed Ismail 13 33 , Urmas Kõljalg 36 , Paul-Emile Lagneau37, 3 , Xingzhong Liu10, Lorenzo Lombard , Wieland Meyer , Andrew Miller , Mohammad Javad Najafzadeh , Lorelei Norvell 13 36 , , William Quaedvlieg 1 , Johan Schnürer , , Bernard Slippers6 , Masako Takashima , Marco Thines , Ulf Thrane , Alev , Bevan 13 , Neriman Yilmaz , Andrey Yurkov , and Ning Zhang

http://www.biology.duke.edu/fungi/mycolab/primers.htm

Barcoding consortium has chosen ITS as primary marker Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi Conrad L. Schocha,1, Keith A. Seifertb,1, Sabine Huhndorfc, Vincent Robertd, John L. Spougea, C. André Levesqueb, Wen Chenb, and Fungal Barcoding Consortiuma,2 a National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20892; bBiodiversity (Mycology and Microbiology), Agriculture and Agri-Food Canada, Ottawa, ON, Canada K1A 0C6; cDepartment of Botany, The Field Museum, Chicago, IL 60605; and d Centraalbureau voor Schimmelcultures Fungal Biodiversity Centre (CBS-KNAW), 3508 AD, Utrecht, The Netherlands Edited* by Daniel H. Janzen, University of Pennsylvania, Philadelphia, PA, and approved February 24, 2012 (received for review October 18, 2011) Six DNA regions were evaluated as potential DNA barcodes for the intron of the trnK gene. This system sets a precedent for Fungi, the second largest kingdom of eukaryotic life, by a multina- reconsidering CO1 as the default fungal barcode. tional, multilaboratory consortium. The region of the mitochondrial CO1 functions reasonably well as a barcode in some fungal cytochrome c oxidase subunit 1 used as the animal barcode was genera, such as Penicillium, with reliable primers and adequate excluded as a potential marker, because it is difficult to amplify in species resolution (67% in this young lineage) (9); however, fungi, often includes large introns, and can be insufficiently vari- results in the few other groups examined experimentally are in- able. Three subunits from the nuclear ribosomal RNA cistron were consistent, and cloning is often required (10). The degenerate compared together with regions of three representative protein- primers applicable to many Ascomycota (11) are difficult to as- coding genes (largest subunit of RNA polymerase II, second largest sess, because amplification failures may not reflect priming subunit of RNA polymerase II, and minichromosome maintenance mismatches. Extreme length variation occurs because of multiple protein). Although the protein-coding gene regions often had introns (9, 12–14), which are not consistently present in a species. MICROBIOLOGY a higher percent of correct identification compared with ribosomal Multiple copies of different lengths and variable sequences oc- markers, low PCR amplification and sequencing success eliminated cur, with identical sequences sometimes shared by several species them as candidates for a universal fungal barcode. Among the (11). Some fungal clades, such as Neocallimastigomycota (an regions of the ribosomal cistron, the internal transcribed spacer early diverging lineage of obligately anaerobic, zoosporic gut (ITS) region has the highest probability of successful identification fungi), lack mitochondria (15). Finally, because most fungi are for the broadest range of fungi, with the most clearly defined bar- microscopic and inconspicuous and many are unculturable, ro- code gap between inter- and intraspecific variation. The nuclear bust, universal primers must be available to detect a truly rep- ribosomal large subunit, a popular phylogenetic marker in certain resentative profile. This availability seems impossible with CO1. groups, had superior species resolution in some taxonomic groups, The nuclear rRNA cistron has been used for fungal dia- such as the early diverging lineages and the ascomycete yeasts, but gnostics and phylogenetics for more than 20 y (16), and its was otherwise slightly inferior to the ITS. The nuclear ribosomal components are most frequently discussed as alternatives to CO1 small subunit has poor species-level resolution in fungi. ITS will be (13, 17). The eukaryotic rRNA cistron consists of the 18S, 5.8S, formally proposed for adoption as the primary fungal barcode and 28S rRNA genes transcribed as a unit by RNA polymerase I. http://fungalbarcoding.org marker to the Consortium for the Barcode of Life, with the possibil- Posttranscriptional processes split the cistron, removing two in- ity that supplementary barcodes may be developed for particular narrowly circumscribed taxonomic groups. ternal transcribed spacers. These two spacers, including the 5.8S gene, are usually referred to as the ITS region. The 18S nuclear

One published indoor microbiome• Amend et al PNAS 2010 “Indoor fungal composition is geographically patterned and more diverse in temperate zones than in the tropics.”• 72 samples of fungi from 6 continents. Sampled ITS2 region and the D1-D2 region of LSU with 454-FLX• Main ﬁnding of increasing species diversity with increasing latitude

Fig 1. Amend et al 2010

ITS 28SPCA of normalized counts – Painted by rRNA type MG-‐RAST tools

PCA of normalized counts – Painted by sampled country MG-‐RAST tools

PCA of normalized counts – Painted by sampled elevaCon MG-‐RAST tools

From barcodes to organisms Dilution to Extinction (d2e)‘High throughput’ isolation from global dust samples Sarea resinae Cryptocoryneum rilstonei Keith Seifert

Summary• New tool development for interacting with genome and metagenome data for Fungi• FungiDB is a resource for genome investigations and repeatable queries and workﬂows• Development of a centralized resource for ITS sequences will enable better analysis of amplicon metagenomics of Fungi

Acknowledgements Marine Biological Lab -‐ VAMPSStajich lab @UCR lab Undergraduates Mitch SoginPeng Liu Jessica De Anda Sue HuseBrad Cavinder Sapphire Ear Anna Shipunova Anthony AmendSoﬁa Robb Lorena Rivera Univ of Colorado at Boulder -‐ QIIME Keith SeifertSteven Ahrendt Carlos Rojas Rob KnightDivya Sain Erum Khan ScoW BatesYizhou Wang Ramy Wissa Gail AckermanYi Zhou Annie Nguyen Jesse StombaughFungiDB Programmers Argonne NaConal Lab -‐ MG-‐RASTDaniel Borcherding Folker MeyerRaghu Ramamurthy Daniel BraithwaiteEdward Liaw Travis HarrisonGreg Gu Kevin Keegan Andreas Wilke EuPathDB @UPenn & UGA David Roos, Jessica Kissinger, Chris Stoeckert Steve Fischer - John Brestelli Brian Brunk - Debbie Pinney Wei Li - Sufen Hu

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Stajich LAMG12 - Indoor Fungi

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