Mycologia, 99(4), 2007, pp. 592–601. # 2007 by The Mycological Society of America, Lawrence, KS 66044-8897 Baudoinia, a new genus to accommodate Torula compniacensis James A. Scott1 sooty, fungal growth, so-called ‘‘warehouse staining’’, Department of Public Health Sciences, University of has been well known anecdotally in the spirits Toronto, Toronto, Ontario, Canada M5T 1R4, and industry for many years. During our investigation we Sporometrics Inc., 219 Dufferin Street, Suite 20C, reviewed a number of internal, industry-commis- Toronto, Ontario, M6K 1Y9 Canada sioned studies of this phenomenon from Asia, Europe Wendy A. Untereiner and North America that attempted to ascertain the Zoology Department, Brandon University, Brandon, taxonomic composition of this material using culture- Manitoba, R7A 6A9 Canada based techniques. Despite the distinctiveness of this Juliet O. Ewaze habitat and the characteristic sooty appearance of the Bess Wong growth, these reports persistently implicated the same Department of Public Health Sciences, University of etiologically implausible set of ubiquitous environ- Toronto, Toronto, Ontario, Canada M5T 1R4, and mental microfungi, chiefly Aureobasidium pullulans Sporometrics Inc., 219 Dufferin Street, Suite 20C, (de Bary) Arnaud, Epicoccum nigrum Link, and Toronto, Ontario, M6K 1Y9 Canada species of Alternaria Nees, Aspergillus P. Micheli ex. David Doyle Haller, Cladosporium Link, and Ulocladium Preuss. A Hiram Walker & Sons Ltd./Pernod Ricard North search of the post-1950s scientific literature indexed America, Windsor, Ontario, N8Y 4S5 Canada by ISI Web of Knowledge failed to yield references to this phenomenon. However a broader search of trade literature and the Web led us to the name Torula Abstract: Baudoinia gen. nov. is described to accom- compniacensis Richon in reference to the warehouse modate Torula compniacensis. Reported originally staining fungus. Subsequently several profitable dis- from the walls of buildings near brandy maturation cussions with Dr Stanley Hughes provided historical warehouses in Cognac, France, species of Baudoinia references key to our rediscovery of this taxon. are cosmopolitan colonists of exposed surfaces sub- In 1872 the French pharmacist Antonin Baudoin jected to large diurnal temperature shifts, episodic brought to the attention of Casimir Roumegue`re and high relative humidity and wetting, and ambient Charles Durieu de Maisonneuve a black, sooty growth airborne ethanol. Morphologically B. compniacensis found on the walls and roof tiles of buildings near resembles some anamorphic Mycosphaerellaceae in distilleries in Cognac, France (Roumegue`re 1881). possessing dark brown, nonseptate or uniseptate Roumegue`re and Durieu de Maisonneuve considered conidia with coarsely roughened walls that are borne the fungus to be an undescribed species of Xenodo- acropetally in unbranched chains and released by chus Schltdl. and informally suggested the name ‘‘X. schizolytic dehiscence. Analysis of partial nuclear baudoinii’’ (Roumegue`re 1881). A few years later rDNA SSU sequences positions B. compniacensis in Adolphe Chatin presented to a meeting of the the order Capnodiales and reveals that it is most Botanical Society of France colonized fragments of closely related to the microcolonial genus Friedman- roofing tiles and stones collected from neighbor- niomyces. Heat resistance is induced by brief sublethal hoods surrounding a distillery in Cognac (fide temperature exposure. Malinvaud 1878). Later the same year Baudoin self- Key words: Capnobotryella, Knufia, microcolo- published a brochure describing the phenomenon in nial fungi, sooty molds, systematics, warehouse which he incorrectly attributed the growth to the staining fungus cyanobacterial genus Nostoc Vaucher (fide Richon and Petit 1881). A more thorough examination of the fungus was undertaken by Richon and Petit (1881), INTRODUCTION who described it as Torula compniacensis. Stanley Several years ago we became involved in a project to Hughes brought to our attention what seems to be characterize superficial fungal colonists on the the only modern investigation of this fungus, a study exteriors of industrial buildings used to house barrels of germination patterns conducted by Kjøller (1961) of spirits during the maturation period. The suscep- published in a source not currently indexed by the ISI tibility of the exteriors of these buildings to dark, Web of Knowledge database. The anamorph genus Torula Pers. : Fr., typified by Accepted for publication 12 April 2007. T. herbarum (Pers.) Link : Fr., was conceived broadly 1 Corresponding author. E-mail: [email protected] to include mostly dematiaceous hyphomycetes with 592 SCOTT ET AL: BAUDOINIA COMPNIACENSIS 593 conidia borne in moniliform chains. Of the more prepared in distilled water and examined in bright field and than 300 taxa that have been included in this genus, Nomarski differential interference contrast microscopy on all but a few since have been redisposed (Crane an Olympus BX-51 microscope. Digital images were 2001). The modern concept of Torula is restricted to acquired with a microscope-mounted Olympus C5050 anamorphs with monoblastic or polyblastic conidio- digital camera. genous cells that give rise to acropetal chains of dark, Thermotolerance.—Stock cultures grown 14 d at 26 C on typically roughened phragmoconidia. The wall of the 90 mm Petri dishes containing MLA were used to prepare distal conidial cell is characteristically thinner and mycelial inoculum for liquid cultures by flooding log-phase lighter in pigmentation than the rest of the conidium, agar cultures with sterile water and gently scraping surface often collapsing proximally to form a cupulate ‘‘co- mycelia into suspension. Suspensions were filtered under rona’’ cell (Crane 2001). axenic conditions through sterile gauze in a thistle funnel covered with aluminum foil to remove large mycelial In his recent nomenclator of Torula, Crane (2001) aggregates. Fragments in the resulting suspension were included both T. compniacensis and T. conglutinata enumerated, their concentration adjusted with sterile Corda var. compniacensis (Richon) Sacc. in Roum. as distilled water to 150 000–250 000 elements mL21 and stored nomina nuda but he did not reassign T. compniacen- at 4 C for up to 24 h until use. sis. Torula compniacensis is not a species of Torula or Thermotolerance in liquid medium of isolates UAMH Hormiscium Kunze, a taxonomic synonym of Torula. 10761 and 10762 was tested by inoculating flasks containing Furthermore it cannot be accommodated in the 50 mL of modified Leonian’s broth (MLB) preconditioned morphologically similar anamorph genera Alysidiella to 26 (control), 37, 42 and 52 C with 200 mL mycelial Crous, Capnobotryella Sugiyama, Cladosporium Link, suspension and incubated at the treatment temperature for Devriesia Seifert & N.L. Nickerson (based on Hormo- 30 min. After incubation cells were harvested by centrifu- dendrum staurophorum Kendrick), Knufia Hutchison gation at 7000 rpm, gently washed with sterile distilled & Unter., Trimmatostroma Corda, or Zasmidium Fr. water, plated on MLA and incubated at 26 C for 14–21 d. Resulting colonies were counted, and the percentage sensu de Hoog (5Racodium cellare Pers.). viability for each treatment was calculated by comparison We have obtained a number of darkly pigmented to controls maintained at 26 C. The thermotolerance assay and slowly growing fungal isolates from environments was conducted in duplicate for 26, 37 and 42 C and in near spirit aging facilities and also commercial triplicate at 52 C. The effect on viability of a 30 min bakeries. Colonies of these isolates are predominantly pretreatment regimen at 37 C before 15 min incubation at subsurface and radiating, producing densely rough- 52 C was tested in duplicate. ened, unbranched aerial chains of schizolytically Thermotolerance under dry conditions also was tested by dehiscing blastoconidia that do not disarticulate harvesting log-phase broth cultures developed at 26 C by readily. The present study describes our investigation vacuum filtration on sterile 25 mm diam 0.8 mm pore mixed of the relationship of our isolates to T. compniacensis cellulose esters membrane (MCEM) filters, drying the cells and morphologically similar taxa based on morpho- at temperatures up to 70 C for periods up to 21 d and reculturing cells on MLA at 26 C. taxonomic methods and the comparison of sequences of the nuclear ribosomal small subunit (SSU) gene DNA isolation and amplification.—Mycelium was harvested and proposes a modern reassignment of T. compnia- from 5 d cultures grown in MLB, rinsed several times in censis. sterile distilled water and stored at 220 C. Axenic culture was verified by incubating a small amount of the harvested mycelium on MLA. Cell walls of 15 mg thawed mycelial MATERIALS AND METHODS aliquots were disrupted by grinding with an equal volume sterile, powdered perlite (Dicaperl Minerals Inc., Socorro, Isolation, culture and storage.—Cultures were obtained by New Mexico) (Scott et al 2000) and total DNA was isolated streaking small amounts of powdered specimens on the in a CTAB-based lysis buffer following the method of Scott surface of modified Leonian’s agar (MLA) (Malloch 1981) et al (2004), modified from Weising et al (1995). Strain solidified with 15 g L21 agar. The medium used for primary information, references and GenBank numbers for taxa isolations was amended with 10 ppm chloramphenicol and used in this study are provided (TABLE I). 5 ppm ethanol after