Mycologia, 00(3), 2008, pp. 417-430. 2008 by The Mycological Society of America. Lawrence, KS 66044-8897

Molecular phylogeny of and other brown rot genera in North America

Daniel L. Lindner Key words: evolution, Fungi, Macro hyporia Mark T. Banik Polyporaceae, loria, root rot, sulfur shelf, Woijiporia (LS.l).A. Forest Service, A!ailisoo lie/il 0/f/ce of ihC exienca Noel/tern Research Station, Center for loreci iVJcoiogi Research, One Cifford Pine/wt Drive, Madison, Wisconsin 53726 INTRODUCTION The genera Laetiporus Murrill, Leptoporus Quél., Pliacolus (Pat.) Pat., P1 cn oporellus Murrill and Woiji- Abstract: Phylogenetic relationships were investigat- poe/a Rvvarden & Gilb. contain that possess ed among North American species of Laeii/ont.s, simple septate hvphae, cause brown rots and produce Lepto1borus, , Pçcnoporellus and Wol/tpona annual, polvporoid I nii ti iig bodies with hyaline using ITS, nuclear large subunit anti iiiitochondrial . These shared morphological and physiologi- small subunit rDNA sequences. Members of these cal characters have been considered important in genera have poroid hymenophores, simple septate traditional polypore (e.g. Gilbertson and hyphae and cause brown rots in a variety of substrates. Rvvardcn 1986, Gilbertson and Rvvarden 1987, Analyses indicate that Laeti/wrus and Wol/ipona are Rvvar(ien 1991) . However recent molecular work I.aeilporhs not monophvletic. All North American indicates that I.ac/iporus, Phaeolus anti Pcnopore11its species formed a well supported monophyletic group kill within the "Ajitrodia dade" of true pOlvpores with (the "core Laeti/orus dade" or Laetiporu.s s.s.) identified by Hibhett anti Donoghue (2001) while the exception of L. persicinus, which showed little I.cptoporus antI Woijiporia fall respectively within the affinity for any for which sequence data are phlehioid" and core polyporoicl" clades of true available. Based on data from GenBank, the southern (Binder et al 200) also fell well hemisphere species L. portentosus Recent molecular and mating studies also have led dade. Woijiporia dilato/cy- outside the core Laetiporus to a revision of genus Laetiporus in North America pha was found to represent a sister group to the core (Banik et al 1998, Banik and Burdsall 1999, Banik and Phaeolus, Prcnopore1lui Laetiporus dade. Isolates of Buidsall 2000) , which formerly contained only Laetiporus dade all fell L. and members of the core sniphureus (Bull.) Murrill and L. Persicinus (Berk & within the Antrodia dade of polypores, while M.A. Curtis) Gilbertson. Burdsall and Banik (2001) fell within Lepioorus Fnoilis and Laetiporus portentosus concluded that genus Lacliporus cocos contains at least six the phlebioid dade of polypores. Wol,tiporia morphologically anti ecologically distinct species. isolates also fell in the Antroclia dade, in contrast to Despite this recent work relationships remain unre- W cocos in the core previous studies that placed solved among North American Lcteliporns species and polyporoid dade. ITS analyses resolved eight clades among other brown rot polyporoid species lacking three of which might represent within Lactiporus s.s., clamps. This study had two goals: (i) to determine undescribed species. A combined analysis using the relationships among North American species of three DNA regions resolved five major clades within Laetiporus and (ii) to determine whether any North a dade containing -inhabiting Laetiporus s.s.: America polyporoid species that cause brown rots and species ("Conifericola dade"), a dade containing L. lack clamps are closely related to Laetiporus. To ("Cincinnatus dade"), a dade contain- cincinnatus accomplish these goals species of Laetiporus, Lepto- L. suiphureus s.s. isolates with yellow pores ing porus, Phaeolus, and Woijiporia were ("Sulphureus dade I"), a dade containing L. sequenced in three regions of rDNA: the intergenic suiphureus s.s. isolates with white pores ( Sulphureus L. gilbertsonii and transcribed spacer (ITS), the nuclear large subunit dade II") and a dade containing (nLSU) and the mitochondrial small subunit unidentified isolates from the Caribbean ("Gilhertso- (mtSSU). nii dade). Although there is strong support for Of the genera included in this study the most groups within the core Laetiporus dade, relationships widely known is Laetiporus. As it is currently defined among these groups remain poorly resolved. Laetiporus includes annual polypore species that produce a brown rot, have dimitic binding hyphae Accepted for publication 26 February 2008. Corresponding author. E-mail: [email protected] and lack cystidia and clamp connections (Gilbertson

417

418 MYCOLOGIA

and Ryvarden 1986, Ryvarden 1991). With the gilbertsonzi occurs in the southern and western regions exception of L. perszcinus, all North American of North America and occurs on a wide range of Laetzporus species produce brightly colored, conspic- hardwoods including . The upper pileus uous fruiting bodies that have been regarded tradi- surface of L. gilbertsonii is generally more salmon to tionally as part of the L. suiphureus s.l. species pink than other Laeliporus species while the pore complex. Species in the L. suiphureus si. complex layer is either yellow or white, which differentiates the are popular edibles that frequently are collected two color forms, L. gilbertsonii with a yellow pore layer under the common name Sulfur Shelf" or "Chicken and L. gilbertsonii var. pallidus with a white pore layer. of the Woods" (Arora 1986). Luetiporus suiphureus s.l. Laeliporus persicinus is the one North American also has been investigated for novel antimicrobial and Laetiporus species not considered part of the L. medicinal compounds (Turkoglu et a! 2007) and its suipliureus s.l. complex. This is due to striking bright pigments have been examined for potential as macroscopic differences, including a brown to pink- food colorants (Davoli et al 2005). Various authors ish-brown pileus surface, it pinkish-cream pole layer from the late 19th and early 20th centuries described and flesh and tubes that bruise blackish-brown. additional varieties and species within the L. sulphur- Laetiporus persicinus originally was described by ens s.l. complex to account fr the wide range of Berkeley and Curtis in 1872 as Poiyporus persicinus morphological and ecological variation exhibited by but was transferred to Laeliporus in 1981 by R. this species, although few of these names were used Gilbertson(Gilbertson 1981). Laetiporus jiercicin us is widely or consistently. Biirdsall and Banik (2001) found in the southeastern United States occurring recognize five species and all variety within primarily oil although it is found occasionally the L. suiphureus si. complex: L. (incinnatus (Mor- on . Burdsall and Banik (2001) hypothesized gan) Burds., Banik & T.J. Volk, L. eon[erieoia Burds. that L. persicinus might not be closely related to & Banik, L. giIbeitsonii Buns., L. gilbertsonii var. species in the L. sulphnreus si. complex based on the pallidus Burds., L. huroniensis Btirds. & Banik inid L. dark pigmentations in the fruiting bodies, the su Iphu reus s.s. appearance of the binding hvphae and preliminary With the exception of Laetiporus rinciii nalus, all RFLP data. species in the L. suiphureus si. complex typically Work by Hibbett and Donoghue (1995) with produce sessile to laterally substipitate pilei with mtSSU nI)NA sequences indicated that genus Phaeo- bright orange upper surfaces and yellow or white Ins is closely related to Laetiporus snlphureus si. This pore layers (Burdsall and Banik 2001). Laetiporus result has been observed in man y analyses (Hihbett cincinnatus produces centrally stipitate, rosette- and Donoghue 2001, Hihhett and Binder 2002, shaped fruiting bodies with a white pore layer; these Binder ci al 2005), which confirmed that Phaeoius is fruiting bodies arise near the base of large diameter not aligned with the Hymenochaetales despite super- hardwood trees, usually Quercus species, iii the ficial similarities (Wagner and Fischer 2001). The eastern and midwestern United States. In North single North American species of Phaeolus treated by America LaeIporus suiphureus s.s. (as defined by Gilbertson and Rvvarden (1987), P. schweinitzii (Fr.) Burdsall and Banik 2001 and the present study) is Pat., is all root rot pathogen in conifer found in the eastern and midwestern United States ecosystems. Like L. cincinnatus and L. persicinus, P. and often occurs on Quercus species although schweinitzii has a centrally stipitate fruiting body specimens occasionally are found on other hard- found at the base of trees. Unlike Laetiporus species, woods. Preliminary evidence based on a limited it possesses a monomitic hyphal system and produces number of ITS sequences suggests that North hymenial cystidia. Young Phaeolus fruiting bodies are American and European populations of L. suiphureus yellow to orange but become predominantly brown s.s. are conspecific (unpubl data), however further with age (Gilbertson and Ryvarden 1987). comparative work is needed. If species barriers exist Genus Pcnoporellus also contains species with between North American and European populations monomitic, yellow to orange fruiting bodies, leading of L. sulphureus s.s., nomenclatural revision of the Gilbertson and Ryvarden (1987) to suggest a close North American species would be required given that relationship between Phaeolus and Pycnoporellus. L. sulphureus s.s. is typified based on European Gilbertson (1981) included both North American material. species of Pycnoporellus, P. alboluteus (Ellis & Everh.) The recently described species L. coniferi cola and L. KotI. & Pouzar and P. fulgens (Fr.) Donk, in genus huroniensis (Burdsall and Banik 2001) are restricted Phaeolus (Gilbertson 1981). However Gilbertson and to conifers, with L. huroniensis being found in the Ryvarden (1987) placed these species in Pycnoporellus. upper midwestern United States and L. confericola Like Phaeolus both Pen op orellus species grow primar- being restricted to western North America. Laetiporus ily on coniferous wood although neither forms the

LINDNER AND BANIK: L TJPORU AND OTHER GENERA II centrally stipitate fruiting bodies typical of Phaeolus. these genera might be warranted in light of recent Pvcn oporellus alboluteus is resupinate and occurs work on Laetiponic. Many studies have demonstrated primarily in western North America while P. fulgens that nonpolyporoicl genera, including the spathulate is it pileate species found predominantly in eastern genus SparaSsis and numerous corticioid genera, are North America. A distinguishing characteristic of closely related to the genera in this study (Hibbett Penopore11us species is it bright red reaction pro- and Donoghue 2001, Binder et al 2005). This duced by 2% KOH on fruiting bodies; Phaeolus emphasizes the need for future work that includes a sehweinitzii in contrast turns dark brown to black in wide range of species with diverse morphologies as 2% KOH (Gilbertson and Rvvarden 1987). well as species that produce clamp connections. This Species in genus Wol/iporia show microscopic work is a first step toward increasing taxon sampling similarities to species of Laetiponis, Phaeolus and of species that might fall within the Antrodia dade of Pvrnoporellu.s despite producing macroscopic fruiting true polypores while also determining relationships bodies that are thin, restipinate and lacking in bright among species in genus Laetiporus. pigmentations. The two species of W)1/ipOria reported front North America, W. COCOS (F.A. Wolf) Rvvarden & MATERIALS AND ME!! LODS (;ilb. and W dilalohvpha Ryvarden & Gilb., are dimitic and possess greatl y inflated skeletal livphae, a /).\A isolation, I(R and sequencing. —Cultures to be defining characteristic of this genus (Rvvarden sequenced were obtained from the culture collection of- the Center for forest Mycology Research, U.S.D.A. Forest 1991). Although Wol/ipona COCOS also has been known Service, Madison held Office of the Northern Research as W. extenSa (Peck) Ginns, Redhead and Ginns Station, which is housed at the Forest Products Laboratory (2006) proposed to conserve the basionynt Iona COCOS in Madison, Wisconsin. Each culture was grown on potato- due to a lack of acceptance of against Daedalea exten.sa dextrose agar and a small amount of the resulting mycelium, The the nomenclaturally correct name W extensa. without associated agar was prepared for use as template conservation of the epithet COCOS should stabilize the DNA in PCR following the protocol of Volk et al (1996). nomenclature associated with this species and end Two regions of nuclear rDNA and one of mitochondrial years of nomenclatural ambiguity (see Ginns and i-DNA were amplified in PCR. Except where otherwise noted Lowe 1983, Ginns 1984). Woljipona COCO.S is most primer designations are those of White et al (1990). P( R widely known for producing large, edible sclerotia was performed with 5X Green Colaq reaction buffer and that have been referred to as tuckahoes" or Indian GoTaq DNA polvmnerase (Promega. Madison, Wisconsin). bread" in North America (Weber 1929). The sclerotia GoTaq reaction buffer was diluted to a I X working of VU COCOS have been reported from China, usua1l concentration and 0.025 units of GoFaq DNA polvmerasc were added per microliter of reaction volume. Each primer under the name Poria COCOS, where they have been collected and used medicinally (Zhang et al 1997, had it final concentration of 0.2 pM and each clNTl (Piomega, Madison, Wisconsin) had it final concentration Wang et al 2004, Wu et al 2004, etc.). of 200 hiM. Therniocycler conditions were: initial denatur- a monotypic genus in North America I.eptOpOruS is ing at 94 C for 2 nun; 30 cycles of denaturing at 94 C for (Pers.) Quélet. Fruiting containing species L. molls 40 s, annealing at 53 C for 40 s and extension at 72 C for bodies of L. 7nolliS are sessile to effused-reflexed and 120 s; and a final extension step of 72 C for 5 mm. display reddish-purple to purple-brown coloration A section of nLSU rDNA was amplified with primers ((;ilbertson and Ryvarden 1986, Rvvarden 1991). LRI 6/LROR. For most of the isolates a section of ITS was LeploporuS is similar to Ceriporia microscopically, amplified with ITS4/ITS5. Partial ITS sequences from having a monomitic hyphal system and lacking both isolates PR-2 and PR-6326 were amplified with primers clamp connections and cystidia (Gilbertson and 1T53/ITS4, while ITS2/tTS5 and ITS3/ITS4 were userl for Rvvarclen 1986). Geriporia has been delimited from Woijiporia cocos isolates. A section of mtSSU i-DNA was Lepto/mruS based solely on the type of decay, with amplified with ms-I /ms-2. Attempts at amplification of each Ceriporia species producing a white rot (Gilbertson of these three regions were made from cultures of these Laetiporus (7nCinnatus, L. conifeneola, L. gulbertsonii, L. and Ryvarden 1986). LeptoporuS moWs is widespread taxa: var. palilduS, L. huronzenszs, L. persicinus, L. in North America but seems to he collected rarely giibertsonii suiphureus s.s., Leptoporus moths, Phaeolus schweznitzzi, based on the number of collections in herbaria. Pvcnoporellus alboluteus, P. fulgens, Woifiporia COCOS, W Although the genera examined in this study are dilatophya and the unidentified isolates PR-2, PR-6326, PR- known to be a polvphyletic assemblage, the y were 914, GDL-1, EUC-1 and KOA-l. Preliminary morphological chosen as a starting point in the search for species data placed all unidentified isolates in genus Laetiporus. that might be closely related to Laetiporus. The goal Collection information for all isolates is provided (TABLE I). of this study was not to complete an exhaustive PCR products were purified with QlAquick PCR Purifica- evolutionary investigation of all of these genera but to tion Kit (QIAGEN) according to the manufacturers determine whether a realignment of species within protocol. Sequencing reactions were performed following

420 MY:OL0GIA

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LINDNER AND BANIK: LAl:TJPORU ANI) OTHER GENERA 421

the BigDye terminator protocol (ABI Prism). The purified a- — o- PCR products for each isolate were sequenced from both C C C Ca- C! C! C! C! C! C! C! the 3 and 5 direction with the same primers used in the C C C C C C Ca-- C PCR amplification. Sequencing products were analyzed at the University of Wisconsin Biotechnology Center (Madi- son, Wisconsin) with air Prism 377 sequencer. z Complementary sequences for each isolate were compared a C C cc 0- C a- a- •: a: - C! — — — and discrepancies resolved with the sequence electrophe- -•: .c r cJ C! C! C! C! C! C! C! C! rograms. For analysis purposes all sequences were termi- C C C C C Ca-- - nated at the base immediatel y adjacent to the primer -4 attachment point. In addition to the primers used in a amplification two primers were synthesized: FPLms-4 (T(;TAC:TTTAGc;TCCTAAA) and FPLms-7 (ACAG x C — C! C a- e LC a- x cc C CC C Cc C Ln -S (;\CTCC(;ACTTCC) These primers were used to se- C! C! C! C! C! C! C! H a- a- quence long mtSSU amplification products that occurred in L. peiainus, 1. fulgens and two of three isolates of 1V. (1/ia /o/rp/l (I. JIrtlogenclic analysts.—Sequeuccs were aligned manually with Sequence Alignment Editor (Sc-Al) v2.0a9. Sequences were deposited in GenBank (see T\IttE 1) and sequence aliginneuts for nLSU, mtSSU, ITS and a combined dataset of all three regions were deposited in Treehase (accession # S1994). Maximum parsimony was implemented in PAUP 4.01)10 (Swofford 2002) and Bayesian analysis in MrBaves 3.1.2 (I Iuelsenbcck and Ronquist 2001, Rouquist and Huelsenbeck -S 2003). a a cc •.!4 H The u [.SU sequences were aligned with 15 additional a sequences from GenBank: A niradia (arhon 00 AF287844, < a Ln bC Ceriporia PulP urea AF287852, Dw,yahoins sodans A\293 176, CS - Fames /omentarius AF287857, Gr!Joia frondo.sa AY629318, G. Jsondo.sa AY826982, Laetiporus por/entosus AY293191, Par- j mast omt ces Ira nsmu tans AF5 18635, IIen rotus ostreatus U04140, Jol-tpon1s squamosus AF393069, ,Sareodon inthrlca- C COCC ins AY5867 I 1, .Sparassis crispa AY2 18386, S. spat/zulata AF287889, S. .cpathulata AY218396 and Woijiporia cocos A17393081. The sequences from Genbank represent a selection of species from the three major polvporoid clades liIi!1i1 (Antrodia, phlebioid and core polyporoid) defined by Binder et al (2005), as well as two sequences from outside the polyporoid dade, which were included as outgroups. Sequences of uI,SL from two isolates originally identified as Laeliporu.s persicinus (PR-2 and PR-6326, [oisporns taipae" C! CccC Ca- in TABLE I) were excluded from anal yses because they could z not he aligned with other species: BlAST analyses of these - C! C! a-- sequences produced no significant matches. S For nl,SU data, heuristic searches were conducted in PAUP with characters unordered and of equal weight and gaps treated as missing data. Default settings were used with these exceptions: stepwise-addition option was set to random with 100 replicates, steepest descent was used with the 11W branch swapping option and the number of trees saved was set to automatically increase. Bootstrap support for clades (Felsenstein 1985) was estimated from 1000 heuristic searches with the same settings described above, C S with the exception that the stepwise-addition option was set to random with 50 replicates. Bayesian inference was - implemented in MrBayes using default settings with the GTR model. Two million generations were performed with samples taken in increments of 100. The first 5000 trees

422 MYcoLo(;I

(25%) were considered the burn-in and were excluded from obtained from 51 isolates (TABLE I) yielding 30 construction of the Consensus tree. After running the sequence variants. All ITS sequences were 555- analysis the standard deviation of the split frequencies was 72() hp long except for WolJipoiia cocos, which had examined to confirm it was below 0.01, the potential scale reduction factor was examined to confirm all parameters an ITS length of 1863 bp. The extreme length of W. were close to 1.0, and the plot of the generations versus the cocos ITS was due to an approximately 1000 bp log probability of the data (the log likelihood values) was insertion in ITS 1 and an approximately 270 bp examined to confirm it had reached the stationary phase insertion in ITS 2 when compared to Phaeolus (was neither increasing nor decreasing). Three indepen- schweinjtzu. dent Bayesian runs were performed and posterior proba- Sequences of nLSU were obtained from 35 isolates bilities were averaged across runs. (T\B1.t; I) yielding 20 sequence variants. Sequences of For the ITS region, sequences from species in the core nLSU were 662-734 hp. Sequences of mtSSU were Laelipnois (lade (sec Fl(;. 1) were found to align easily with obtained from 31 isolates; Wo(/ipooa dilatohyp/ia and sequences, while sequences from all isolates PR-2 and PR-6326 did not yield mtSSU other species were found to be more than 25% dissimilar based on sequence identity, making them difficult or sequences. Most mtSSU sequences were 491-610 bp. However L. persicinns, P. fulgent and two of three impossible to align. Analyses for the ITS region therefore isolates of 14g. were pc-formed with species in the core Laetipou-us dade dtiato/rpha had large insertions that with W. di/atohyplia as olltgroup. Ten I.aelzporus .i/p/uuirus resulted in sequence lengths in excess of 1700 bp; ITS sequences from (;coBink (AF229 196, AM269785, these insertions occurred at the sante location within AM269786, AY089742, AY218417, AY835667, AY8356681 each sequence. Insertions were removed to align DQ22I 108, DQ450876 and FF088657) were found to align sequences. (GenBank accession numbers can he with our dataset and were included in analyses. Maximum found in TABLE I.) parsimony and Bayesian analyses were run on the ITS sequences using the methods described for nLSU data with Phylogenetic anaivses.—The heuristic search using these exceptions: 1 400 000 generations were run in the nLSU sequences produced 44 equally parsimonious Bayesian analysis and the born-in was set to 3500 (25%). trees (one of which is shown in Fl(;. 1) (length = 946, Maximum parsimony and Bayesian analyses were run on Cl = 0.517, RI = 0.757). Five of the seven Laetiporus the miSSU sequences with the methods described for ITS species included in the analysis (I. cincinnalus, I,. data. All species (T,-Bit: I) were included with the exception conifericola, L. gilberlsonii, L. huroniensis and L. of species for which sequence data were not successfully suiphureus) fell in the group referred to as the core obtained (see rsul:rs). Leptoporits mollis was used as Laetiporus dade or outgrotip based on the results of the nLSU analysis. Laetiporus s.s., while two Laeti- porus species (L. ercicinus and A combined analysis was conducted with all isolates for L. orten1osus) fell OtltSi(Ie the core J.aeliporus dade. which complete nLSU, intSSU and ITS data were available. The unidentified - Species were included in this analysis it sequence data could Laetipoi us isolates from Hawaii and the Caribbean be aligned across all three regions. Potential conflicts fell within the core Laetiporu.s- dade. Wol/iporia among DNA regions were identified by first running an djialoh,pha isolates formed a sister group to the core independent parsimony analysis on each of the three 1.aetiporus dade. regions. The parsimony analysis included running a Although a relatively small number of species were heuristic search in PAUP using default settings with these included in the nLSU analysis, the tree reflects the exceptions: stepwise-addition option was set to random with overall structure of the polyporoid dade as defined by 100 replicates, steepest descent was used with the TOR Binder et al (2005), including Antrodia, phlehioid and branch swapping option and the number of trees saved core polyporoid clades. Laetiporus p was set to automatically increase. A strict consensus tree was ersicinus, Phiteolus sc/wejnitzii Pycnoporellus alboluteus, P. fulgens constructed for each region, and the trees were examined and the isolates of for potential conflict. Bootstrap analyses were run for each WoijIporia cocos sequenced for this study all fell within the Antrodia dade. The one dataset with the methods described for the nLSU analysis. W. cocos No conflicts were observed among the datasets (i.e. no sequence taken from GenBank (AF3930811) fell within conflicting branches were observed among the consensus the core polyporoid dade (FIG. I). Phaeolus schweinit- trees and no conflicting branches were supported with a zzi formed a dade with the W cocos isolates sequenced bootstrap value >70), so the three regions were combined for this study, although this dade has weak statistical into one dataset. Maximum parsimony and Bayesian support (bootstrap ^--70 or posterior probability analyses were run on the combined dataset with the -^0.95). Pycnoporellus alboluteus and P. fulgens formed methods described for the ITS region. a dade with strong statistical support (bootstrap 70 and posterior probability ^!0.95). Leptoporus mollis, RESULTS Ceriparia purpurea and Laetiporus portent osus formed a strongly supported dade representing the phlebioid PCR and sequencing—Complete ITS sequences were dade of polypores.

423 LINDNER AND BANIK: LAETIPORUS AND OTHER GENERA

Laetiporus gilbertsonhi var. pallidus TJV2000-i01 J4 Laetiporus gilbertsonhi var. pallidus FP1 50268 It I riiIhrt.cnnii CA-13

Bootstrap support 0 70 and Laetiporus sp. GDL-i (Caribbean) Bayesian posterior probability? 095 Laetiporus sp. PR-914 (Caribbean) C-) 0 50 < Bootstrap support < 70 and TJV99-150 (white pored) Laetiporus su/phureuS CD Bayesian posterior probability 00.95 Laetiporus su/phureus MAS-2 (white pored) Bootstrap support ^: 70 or Laetiporus huroniensis Ml-14 Q) Bayesian postenor probability 0095 CD

Laetiporus huroniensis HMC-3 0 Branch collapses in strict consensus 0 of equally parsimonious trees Laetiporus conifer/cola JAM-1 Laetiporus conifer/cola CA-8 (I) Laetiporus su/phureus CT-1 C-) Q) DA-41 ci Laetiporus sulphureus GR-i2 CD Laetiporus cincinnatus DA-37 Laetiporus cincinnatus 46-1104 Laetiporus sp. EUC-i (Hawaii) Laetiporus sp. KOA-i (Hawaii) 0 Wolfiporia di/atohypha CS-63 Wolfiporia dilatohypha FP72 i 62 C) Wolfiporia dilatohypha FP94089 Q) Q- Phaeolus schweinitzii DA38 M Phaeolus schweinitzii H H B 18924 Wo/fiporia cocos MD-i 06 Wolfiporia cocos MD-275 Laetiporus persicinus RLG14725 Laetiporus persicinus HHB9564 HHB17342 Pycnopore/Ius fulgens FP101689 Pycnoporellus alboluteus HHBi 7598 Pycnoporellus alboluteus H H B 12816 Dacryobolus sudans AY293176 Parmastomyces transmutans AF51 8635 Antrodia carbon/ca AF287844 Grifola frondosa AY62931 8 - Grifola frondosa AY826982 ,arassis spa thu/ata AF287889 :S) arassis spathulata AY2 i 8396 Sparassis cr/spa AY21 8386 Leptoporus mo/I/s TJV93-174 -

Leptoporus mo//is RLG7163 9 Ceriporia purpurea AF287852 Laetiporus port entosus AY293191 10 a. Fomes fomentarius AF287857 "Wolfiporia cocos"AF393081 squamosus AF393069 81 OF) CD - Sarcodon ,mbncatus AY5867 ii P/eurotus ostreatus U04140 - 10 changes

trees (length = 946, Cl = FIG. 1. Analysis based on nl.SU sequences. The tree shown is one of 44 equally parsimonious with 0.517, RI = 0.757). Data were taken from GenBank for isolates not included in TABLE I; these isolates are labeled GenBank accession numbers. All other isolates were sequenced for this study and are labeled with collection numbers. Quotation marks around a species indicate a suspect identification.

The heuristic search with ITS data produced 98 five described species (Laeliporus cincznnatus, L. equally parsimonious trees (one of which is shown in conifen cola, L. gilbertsonii, L. huroniensis and L. FI(;. 2) (length = 191, CI = 0.843, RI = 0.941). Eight suiphureus s.s.) as well as three apparently unde- clades were found, seven of which have strong scribed species of Laetiponus. Relationships among statistical support. These eight clades correspond to Laetiporus species were poorly supported, although 4.4 )I O( .L\

Laet/porus huron/ens/s HMC-i Laet/porus huron/ens/s HMC-2 Laetporus huron/ens/s HMC-3 Laetiporus huron/ens/s Ml-7 Laetiporus huron/ens/s Laet/porus huron/ens/s MI-14 Laetiporus con/fer/cola JAM-1 Laetiporus conifer/cola NV-2 Laet/porus conifer/cola TREE-5 Laet/porus conifer/cola Laet/porus conifer/cola CA-8 Laetiporus conifer/cola AK-1 Laet,porus "suiphureus DQ450876 Laet,porus "sulphureus" AY835667 Laet,porus suiphureus EF088657 U nknown Laetiporus sp. (GenBank) Laet/porus "sulphureus" AY2 1841 7 Laet/porus sp. EUC-i Laetiporus sp. KOA-1 Unknown Laetiporus Sp. (Hawaii) Laetiporus g//bertson// CA- i 3 Laetiporus g/Ibertson// OR-2 Laetiporus "suiphureus" AM269785 Laetiporus gI/bertson// var. pa/lidus L-1 2954 Laetiporus gilbertsoni, Laetiporus g/Ibertson/, var. pa/I/this FPi50268 Laet/porus g//beson// var. pal//dus TJV2000101 Laefiporus sp. GDL-1 Laet,porus sp. I Unknown Laetiporus PR-914 I sp. (Caribbean) Laetiporus cincinnatus DA-37 Laet/porus c/ncinnatus MAS-1 Laet/porus clnc/nna(us ws-i Laetiporus cincinnatus Laetiporus c/nc/nnatus 46-1104 Laetiporus "sulphureus" AM269786 Laetiporus su/phureus CT-1 Laet/porus suiphureus DA-41 Laetiporus suiphureus GR-12 Laet,porus suiphureus NJ-2 Laet/porus suiphureus AF2291 96 Laetiporus sulphureus MAS2 Laetiporus su/phureus s s. Laetiporus su/phureus DQ22 1108 Laetiporus suiphureus TJV99-150 Laet/porus sulphureus AY089742 Laetiporus su/phureus • Bootstrap support^ 70 and AY835668 Bayesian posterior Probability 9095 Laetiporus su/phureus TJV95-84 .50 Bootstrap support 70 and Laetiporus suiphureus ERT-713 Bayesian posterior probability ^ 0 95 VVolfipona di/atohypha CS-63 Wo/fipona di/atohypha 0 Bootstrap support 2--! 70 or FP72 162 Bayesian posterior probability^ 0 95 L4o/f/pona dilatohypha FP94089 Branch collapses in strict consensus - 5 changes of equally parsimonious trees

FIG. 2. Analysis based on ITS sequences. The tree shown RI = 0.941) is one of 98 equally parsimonious Data were taken from GenBank for trees (length = 191, CI = 0.843, accession isolates not included in TABLE I; these numbers. All other isolates were sequenced for this study isolates are labeled with GenBank and are labeled with collection around a species indicate a suspect identification. numbers. Quotation marks

there is strong support for a dade containing L. isolate collected from gilbertsonji and the apparently undescribed Caribbean QUeTCUS in northern Germany, isolates. There is moderate support for a dade clustered with North American isolates of L. sulphur- Containing eus s.s. L. conifer/co/a and L. huron/ens/s. Two sequences (AM269785 and AM269786) listed in Results from the heuristic search with mtSSU sequences produced few GenBank as Laetiporus suiphureus statistically Supported clades. clustered strongly The core with other Laetiporus Laetiporus spp. (L. gilber/sonji and L. dade was weakly supported, while cincinnatus respectively). The GenBank sequence relationships within the dade were unresolved Alignment of AY835668, which is derived from a mtSSU sequences from species outside L. suiphureus the core Laetiporus dade was difficult due to large

LINDNER ANI) BANIK: LAb,-rjioi?uS AND OTHER GENERA 425

variable regions; alignment within the core Laetiporus patterns. The two isolates of this type included in dade showed little variation among isolates, with the this study (MAS-2 and TJV99-150) cluster with L. exception of one variable region approximately 30 suijihureus s.s. in the ITS analysis (Fl(;. 2) and are base pairs long. Within this variable region a 16 bp clearly differentiated from both L. cincnnatus and L. mdcl (TTTAATTTTAAATTCA) was found to occur in gilbertsonii var. pallidus, the two species that are four isolates: two L. (on/fericola isolates (JAM-i and known to possess white pore layers.. ITS data indicate CA-8) and two L. cuiphureus s.s. isolates with white that these white-pored isolates fall within L. silphur- pores (MAS-2 and TV99-150); this mdcl did not eus s.s., yet both the nLSU (Fl(;. 1) and combined occur in any of the other isolates sampled. datasets (Fl(,. 3) support separation of white pored The heuristic search with the combined dataset forms of L.S uiplwreus s.s. from isolates with yellow produced 31 equally parsimonious trees (one of pores. Additional isolates are needed to determine which is shown in FR;. 3) (length = 244, CI = whether this (lilierentiation warrants the description 0.812, RI = 0.892). Five clades were resolved with of a new species or variety. Uiihrtunately single weak to strong statistical support. Laetiporu.s gilberiso- spores from white-pored L. sul/thureus s.s. fruiting nii isolates and the 1.aetiporus isolates from the bodies have low germination rates across a wide pH Caribbean formed a weakly supported dade (Gilbert- range of agar media (unpuhl data), which has sonii) . Laetiporns (/il(iflflaIuS isolates formed a hampered mating studies. Although names have been L. strongly supported dade (Cincinnatus) that did not introduced to describe white-pored forms of group consistently with any other species (the branch suiphureus sI., such as Pecks L.s ulphureu ar. containing Gilhertsonii and Cincinnatus clades col- scinjaihinus (Peck 1906), it is likely these names lapses in a strict consensus tree). Laeiiporus sulphur- describe the more common and morphologically (Birdsall and Banik eus s.s. isolates split into two strongly supported distinct species L. cjncznnatus clades, one with yellow pores (Sulphureus dade I) 2001). and one with white pores (Sulphureus dade II). The The only other Laeliporits Species found to have er was I.. two conifer-inhabiting species, L. conifericola and L. variation in the color of the pore lay I.. giibertsonii are huron iensis, formed a weakly supported dade (Con- gilhertsonii. Both color forms of ifericola) . The three branches indicating relation- almost indistinguishable in these anal yses, with only ships among major clades collapse in a strict two base pairs of variations in both the ITS and nLSU consensus tree. regions. This suggests that a change in pore colora- tion can occur with little additional genetic differen- tiation in this group. Laetiponic gilhertsonii also shows t)IS( ISSI()N variation in host preference, being found on both Sequence analysis confirms that Laetzponis sulthUreUS Quercus spp. and Eucalyptus spp. (Burdsall and Banik s.1. represents a large species complex in North 2001). The only other isolate in this study collected America, which supports work by Burdsall and Banik on Euca1ptus, EUC-1 , was collected in Hawaii and (2001) using mating compatibility, RFLP, morpholo- clustered with an addition Hawaiian isolate from a gy and ecology. Although five major clades encom- native Acacia koa (koa tree). These Hawaiian isolates, passing eight species groups were identified within along with the Caribbean isolates in the Gilbertsonii Laeiiporus s.s., further work is needed to resolve dade and the white-pored isolates of L. sulphureus relationships among species and to determine the s.s., represent three likely candidates for undescribed number of Luetiporus species in North America. species in the core Laetiporus dade. Lae/iporus sulDhureus s.l. is known from many A fourth undescribed Laetiporus sp. was identified continents, including Africa, Asia and Europe (Ry- in the ITS analysis (FI(;. 2, L. suiphureus" AY2I 8417, varden and Johansen 1980, Rvvarden and Gilbertson AY835667, DQ450876, EF088657), although all rep- 1993, Nüñez and Rvvarden 2001) so additional resentatives in this group are GenBank sequences for species undoubtedly remain uncharacterized world- which little geographic or host data exist. One wide. sequence of this type (AY835667) was obtained from Of all species in the core Laetiporus dade, a Laetiporns suiphureus sI. isolate collected on Picea Laetiporus suiphureus s.s. displayed the largest num- abies in southern Germany (Davoli et al 2005), raising ber of sequence variants and also showed variation in the possibility that these isolates are conspecific with pore color, either white or yellow. Burdsall and Banik the European conifer-inhabiting Laeti/iorus sulphur- (2001) describe Laetiporus suiphureus s.s. as possess- ens sI. isolates investigated by Rogers et al (1999). ing lemon yellow pores but mention rare specimens Burdsall and Banik (2001) report that L. monticola with white pores that are indistinguishable from L. Cerny. was described in 1989 to accommodate suiphureus s.s in morphology, habitat and RFLP European Laetiporus specimens found on conifers,

hk r

426 Mycot,o;i>

Laetiporus gilbertsonii var. pallidus TJV2000-1 01 • Bootstrap support 0 70 and Bayesian posterror probability >095

50 < Bootstrap support < 70 and G) Bayesian posterior probability 0.95 Laetiporus gilbertsoni/ var. pallidus FP1 50268 0 CD Bootstrap support ^: 70 or Bayesian posterior probability 0.95 Branch collapses in strict consensus Laetiporus gilbertsonii CA- i 3 of equally parsimonious trees :

Laetiporus sp. PR-914 (Caribbean)

Laetiporus sp. GDL-i (Caribbean)

Laetiporus cincinnatus 46- 1104 Ic) 0 I Q- — Laetiporus cincinnatus l CD DA-37 I E C)

Laetiporus suiphureus CT-1 (I)

Laetiporus suiphureus GR-i2

cn Laetiporus suiphureus DA-41

Laetiporus sulphureus MAS-2 C,)

Laetiporus suiphureus TJV99-1 50

Laetiporus conifer/cola JAM-1 C) 0 Laetiporus conifer/cola CA-8 FG (D0 0 ci) Laetiporus huroniensis MI-14

Wo/fipor/a dilatohypha FP94089 White pores

Wo/fiporia dilatohypha FP72162 E3 Yellow pores Eli Angiosperm hosts Wolfiporia dilatohypha CS-63

- 10 changes E Gymnosperm hosts

FIG. 3. Combined analysis bascd on nLSU, mtSSU and ITS sequences. The tree shown is one of 31 equally parsimonious trees (length = 244, Cl = 0.812, RI = 0.892). Isolates were included in this analysis only if sequence data were available for all three regions.

although apparently this species was never validly constitute an unrecognized species. GenBank se- published due to a lack of a Latin diagnosis. Further quence AY835668, which is derived from a L. taxonomic work is needed to determine whether suiphureus isolate collected on Quercus in northern conifer-inhabiting Laetiporus isolates from Europe Germany (Davoli et al 2005), clustered with North

LINDNER AND BANIK: LAP,TIP01?US AND OTHER GENERA 427

American L. suiphureus s.s. isolates (Flu. 2), support- closely related to any of the species in the core ing the hypothesis that L. suiphureus s.s. is restricted Laetiporus dade. Although L. portentosus was placed to angiosperms and is found in both North America in Piptoporus, Rajchenherg (1995) transferred it to and Europe. Laetiporus based oil and cultural GenBank ITS data also produced two sequences of characteristics. Unfortunately these characters appear Laetiporus sn Iphu ren.s (AM269785 and AM269786) to he phylogenetically uninformative at the generic that clustered strongly with known Laetiporus isolates level for these species groups. Hibhett and Dono- identified independently with mating compatibility ghues (2001) analysis indicated that L. Porlentosus and fruiting body morphology (Banik and Burdsall falls within the "Antrodia dade" of true polvpores, 2000, Burdsall and Bariik 2001). Laetiporus sulphur(,us although the nLSU analysis in this study placed it in sequence AM269785 is from a Californian isolate the phlebioicl dade. More gene regions from addi- associated with an unknown host (Guglielmo et al tional L. porlentosus isolates need to be sequenced to 2007) and clustered with L. giibertsonu sequences; L. resolve this issue. "sulphurcus sequence AJ\4269786 is from an isolate Like Lactiporus portentosus, L. pers/cinus falls collected oil in Wisconsin (Guglielmo ci al outside the core Luetiporus dade in the nLSU 2007) and clustered with L. (jncnnalns sequences analysis. This result is consistent with L. persicinus (FI(,,. 2). The phylogenetic placement of AM269785 divergent morphological traits, including a lack of and AM269786, taken together with the host and bright orange coloration in the pileus surface and geographic data associated with their parent isolates, flesh and tubes that stain blackish-brown. BLAST indicate the species names aSSociatedl with these analyses of the GeneBank (NCBI) databases using GenBank sequences are in need of revision. ITS, nLSU and mtSSU regions from L. persiclnns The discovery of Woijiporut dilatohpha as a sister revealed no significant similarity to any known genus, group to the core Laetiporus dade was fortuitous and suggesting that placement in a new genus may he might help to define the generic limits of Lactiporus warranted. However further work is needed to as sequence data accumulate from other regions of confirm that ail genus does not already the world. Two species of Laetiporus, L. portentosus exist. Since being described as Polvporus pdrczcinus by and L. perczcinns, fell well outside the core Lae/iporus Berkeley and Curtis in 1872, this species has been dade, indicating that Laeliporus as it is currently placed in numerous genera, including Meripilus defined represents a polyphyletic assemblage. The (Ryvarden 1972) and Buglossoporus (Corner 1984), other North American polypore species sequenced before being transferred to Laetiporus by Gilbertson for this study, including Leptoporus mollis, J-haeo1us (1981). schwein itzii, Pycn oporellus albolu teus and P. fu lgrns, W0Uiporth dilatohypha, which formed a sister group also do not appear to be closely allied with the core to the core Laetiporus dade, was the only species Laetzporus dade; however, with the exception of L. outside genus Laetiporus with ail sequence that rnollis, all these species fall in the Antrodia dade of aligned easily with core Laetiporu.s species. Woijiporia polypores. The Wol/iporia cocos isolates sequenced for dilalohypha isolates were so similar to species in the this study also fell within the Antrodia dade, although core Laetiporus dade that future work must address this is in contrast to earlier studies that placed W whether W. dilatohypha is congeneric with species in cocos in the core polyporoid dade (this point is the core Laetiporus dade. The Woijipona cocos isolates addressed in detail later in the discussion). Further sequenced for this study were found to be distantly taxon sampling and additional sequences analyses are related to both W dilatohypha and all Laetiporus needed to confirm the placement of these brown rot species examined. This indicates that Wolfiporia, like polypore species in broader l)asidiomycete phyloge- Laetiporus, is polyphyletic. Although five species are nies. presently found in genus Woifiporia, only two were A lack of monophly in Laeliporus first was demon- included in this study. Further taxon sampling is strated by Hibbett and Donoghue (2001) with needed to determine whether any of the three sequence analysis of two rDNA regions from the remaining Woijiporia species, W cartilaginea Ryvar- southern hemisphere species L. portentosus. Using a den, W. curuispora Y.C. Dai, and W. suiphurea (Burt) single isolate of L. suiphureus s.l., Hibbett and Ginns, are closely allied with other Woijiporia species Donoghue (2001) found that L. portentosus is not or genus Laetiporus. closely related to L. suiphureus s.l. Although both L. The close relationship between Laetiporus and portentosus and L. persicinus show macroscopic, Wolfiporia dilatohypha was surprising given that W microscopic and physiological similarities to other dilatohypha fruiting bodies are macroscopically unlike Laetiporus species, sequence data suggest these are Laetiporus fruiting bodies. Wolfiporia dilatohvp hue convergent characters and that neither species is fruiting bodies are effused, thin (3-4 mm) and white

428 M\tOLOGIA

U) 1)1111 (oIu1c(I with iclali\cI\ hugc ImIcs (1-7) such an analysis might be hampered by the size of the mm), which is in contrast to the large, brightly ITS region in W COCOS, which was found to be greater colored, pileate fruiting bodies of Laetiporus. Three than 1800 base pairs due to insertion events. In cultural isolates of lIV. dilato/cpha were sequenced for addition to making the ITS difficult to amplify, these this study and all had identical LSU sequences and insertions make it difficult to align the ITS of 14 7. COCoS nearly identical ITS regions (two base pairs of with other species. variation). However no sequences have yet been Although previous investigations have suggested obtained from fruiting body material and the most that Phaeolus schweinitzii is closely related to Lae/i- recent cultural isolate was collected in 1963. Fresh p01715 sulp/wreus s.l., the ITS of P. sChzleinitzij is fruiting bodies of W di1atohp/ia are needed to difficult to align with Laetiporus spp. and the nLSU of confirm that the sequences iii this study are consistent I. sChweznz zji shows only 89% sequence identity to L. with sequences derived from fruiting structures. suiphureus s.s. The closest relative of P. sr/iweinitzii in Unfortunatel y fruiting bodies of this species seem to the nLSU anal ysis was Wo/iporia COCOS, although this he rare or are rarely collected. finding has weak statistical support. Further sampling Although Wofiporia dilaio/e5p/ia was found to be is needed to determine whether close relatives of I. closely related to the core Laeiiporus dade based on SC/Iue/njfzf/ exist and whether there is variability ITS and nLSLf data, it is interesting to note that within this widespread species. The proposed link isolates of W COCOS, the type species of Wol/iporia, do between I/iaioiu.s and PyCnoporellus (Gilbertson and not appear to he closely related to VV. (iz1atohpIia. Ryvarden 1987) was not borne out by this study The two W COCOS isolates sequenced fr this study because Jhaeoius sChzoejlI/IZU and PyCnoporeilus iso- (MD-106 and MD-275) both had identical ITS, nLSU lates shared only 85% sequence identity for the nLSL and mtSSU regions. An additional 13 isolates from and alignments of ITS regions were impossible the Center for Forest Mycology Research (CFMR) between these genera. The two PCnoporei/us species cultural collection (Madison, Wisconsin) were se- however showed similar nLSU sequences (97% quenced in the nLSU region and all displayed sequence identity) and the ITS regions were aligned sequences that matched the two original isolates easily. This evidence, together with the similar almost identically (unpubl data). However nLSU and microstructures and chemical staining reactions in mtSSU data for 147. COCOS in this study differ strongly KOH, suggests that Pycnoporellus consists of a natural from those reported by Hibbett and Donoghue grouping of species. (2001) and Binder and Hibbctt (2002). Hibbett and Unlike the other genera investigated, genus Lepto- Donoghue (2001) sequenced the nuclear and mito- poruc seems to fall outside the Ai1trodia dade as chondrial SSU of W COCOS (isolate No. FP1,4198), defined by Binder et al (2005). The nLSU sequences while Binder and Hibhett (2002) sequenced the of LetoporuS generated for this study match almost nuclear and mitochondrial LSU of the same isolate identically (99% identity over 643 base pairs) those of (the GenBank nLSU sequence of this isolate is Binder et al (2005), who found that Leptoporus fell included in FIG. 1 as "Woijiporia COCOS" AF393081). near (]eriporia in the phlebiod dade. Genus Lepto- Sequences from isolate FPL4198 subsequently have porus and Geriporia share similar microscopic charac- been included in many analyses by various authors ters and Gilbertson and Rvvarden (1987) state that (Hibbett and Binder 2002, Hihhett 2004, Binder et al the production of brown rot is the only differentiating 2005). Isolate FPL4198 has shown strong affinities lot- character for Leptoporus. If this placement is correct it the core polypore dade, clustering with white rot would indicate that Leptoporus arose within a dade of genera such as Polyporus and Trametes (see FI(,,. 1). white rot species. Isolates of Leptoporus were found t Kim and Jung (2000) however sequenced the display two ITS types that differed by 18 base pairs, nuclear SSU of W. Cocos (isolate ATCCI3490) and indicating that further work is needed to determine found that it clustered with other brown rot genera whether L. moths represents a species complex. such as Lactiporus, Phaeolus and Sparassis. As pro- In this study two isolates (PR-2 and PR-6326) were posed by Binder et al (2005), this disparity might be encountered that possessed extremely divergent due to the mislabeling or contamination of isolate nLSU sequences. These isolates came from collec- FPL4198, which does not appear to represent W. tions that had been classified as Laeliporus persicinus, COCOS. Unfortunately the original isolate of FPL4198 although preliminary sequence analysis indicated housed at CFMR died in culture and is not available these isolates varied strongly from L. Persirinus and for further study. Additional cultural isolates, fruiting all other species included in this study. Although ITS bodies and scierotia of W. COCOS are needed to clarify data could not be obtained, the nLSU region from the position of this species and to determine the these isolates was amplified successfully from cultures extent of variability within this species. Unfortunately and fruiting bodies and had no significant BLAST

LINDNER AND BANIK: LAETJPORLTS AND OTHER GENERA 429

matches in GenBank. Preliminary parsimony analysis phylogenetic analyses and helpful advice oil places this species in the core polyporoid dade, versions of this manuscript. although an insertion of approximately 150 base pairs makes alignment of these sequences difficult (Karl- LITERATURE CITED Henrik Larsson pers comm). This species has dark y, Califor- brown fruiting bodies that grow as basal rosettes from Arora D. 1986. Mushrooms demvsmified. Berkele Press. 959 p. and nia: Tell tree roots iii a fushion similar to L. persicinus Banik MT, Burdsall HI Jr. 1999. Incompatibility between L. persicinus or Phaeolus scI/weinllzi; however, unlike Laetiporus uncin na/os and L. u/pliureuc in culture. P. schweinilzii, the fruiting bodies can attain tremen- Mvcotaxon 70:461-469. dous dimensions (0.5-0.8 m diarn) and have a thick, 2000. Incompatibility groups among North black pileus cuticle. Preliminary evidence based on American populations of Laelzponts suiphureus sensu excavating tree roots close to fruiting bodies suggests lato. Mvcotogia 92:649-655. this species produces a white rot rather than the Volk 1]. 1998. Identification of groups brown rot associated with L. pecciewus (1). Jean Lodge within Laetiporus .solphureus iii the United States based pers comm). The nLSU data confirms that this on RFI.P analysis of the nuclear ribosomal DNA. Folia species is highly divergent from all other polvpore Crvptog Estonica 33:9-14. species for which sequence data are available despite Binder NI, I tihhett DS. 2002. Higher-level phvlogenetic relationships of I Iomobasidiomcetes (mushroom- its morphological anti microscopic similarities to L. forming fungi) inferred from four rDNA regions. Mol persicin us. Gross morphology suggests this taxon Phvlogenet Evol 22:76-90. might represent Ioiporus Ia/Jute Cooke ( = Menpi/us 1,arssoti KH, Larsson E, Langer E, Langer Ialpae [Cooke] D.A. Reid) or possibly Men pt/us G. 2005. The phvlogenetic distribution of resupinate iroJncalis Guzmán & Pérez-Silva (Guznián and Silva forms across the major clades of mushroom-forming 1975). This species also might have been described by fungi (1-lomnohasidioiuycetes). Svst Biodivers 3:113-157. Murrill from Jamaica in 1910 (Murrill 1910) as Biirdsall HHJr, Banik MT. 2001. The genus J.aetlporo S Ill A 7nauroderina /irtllonii Murrill. Both Polporus Iaipae North America. Harvard Pap Rot 6:43-55. and Amauroderma brittonii long have been considered Corner EJH. 1984. Ad Polyporaceas III. Pzpto/..orus, Bu/,ios- and Bondatzewia. Bei- synonyms of Laeii/.ionts persicin us but further work is sojx)rus, Luetiporus, Menpilns, needed to determine the legitimacy of this synonym. hefte br Nov Hedwig 78:137-222. Davoli P, Mucci A, Schenetti L, Weber R. 2005. I.aetiporie It is clear that many species within the Antrodia acids, it family of non-carotenoici polyene pigments dade of polypores remain poorly characterized. front and liquid cultures of Laetiponis Additional isolates of Laetiponts, Leploporus, Phaeo/us, sulp/inreu.s (Polvporales, Fungi). Phvtochemistry 66: Pcnopore//us and Wo/fiporia are needed from wider 817-823. geographic regions to clarify the positions of these Felsenstein J 1985. Confidence limits on phvlogenies: an brown rot polypores in larger basidiomycete phylog- approach using the bootstrap. Evolution 39:783-791. enies. Laetiporus suiphureus sI. in particular requires Gilbertson RI.. 1981. North American wood-rotting fungi further attention oil worldwide basis because this that cause brown rots. M ycotaxon 12:372-416. species complex is one of the larger and more Rvvarden L. 1986. North American Polypores. economically important groups in the Antrodia dade. Vol 1. Oslo, Norway: Fungiflora. 433 p. 1987. North American Polvpores. Vol 2. The sequences provided by this study hopefully will Oslo, Norway: Fungiflora. p 434-885. provide a framework that aids further phylogenetic (;inns J . 1984. New names, new combinations and new and taxonomic investigations. synonymy in the Corticiaceae, Hymenochaetaceae and Polyporaceae. Mycotaxon 21:325-333. Lowe JL. 1983. MacrohyJJOrla extensa and its ACKNOWLEDGMENTS synonym Iorza cocos. Can J But 61:1672-1679. We thank Don Hemimies of the Universit y of Hawaii at Hilo Guglielmo F, Bergemann SE, Gonthier P, Nicolotti G, for providing Laetiporus isolates from Hawaii; D. Jean Garbelotto M. 2007. A multiplex PCR-based method for Lodge of the USDA-Forest Service, Center for Forest the detection and early identification of wood rotting Mycology Research, for providing isolates and fruiting fungi in standing trees.J Applied Microhiol 103:1490- bodies of Laetzporus and 1o1porus talpae from Puerto 1507. Rico; and Thomas j. Volk of the University of Wisconsin at Guzmán G, Silva EP. 1975. Una nueva especie de Meripilus, La Crosse for collecting many of the Laeliporus specimens bongo gigante destructor de la madera en la selva used in this study. We also thank David Hibbett and Zheng tropical de Mexico. Bol Soc Mex Mic 9:53-59. Wang of Clark University for providing sequences of Hibhett DS. 2004. Trends in morphological evolution in Sparassis as well as access to large sequence databases of Homohasidiomycetes inferred using maximum likeli- homobasidiomycetes. We are grateful for the help of David hood: a comparison of binary and multistate approach- Hibbett and Karen Nakasone; they provided assistance with es. Syst Biol 53:889-903. I

430 MYCOLOGIA

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