Fung. Sci. 28(1): 45–53, 2013
Hyphodontia juniperi (Basidiomycota) newly recorded
from China
1 2 3* Eugene Yurchenko , Hong-Xia Xiong & Sheng-Hua Wu
1Department of Biotechnology, Paleski State University, Dnyaprouskai flatylii str. 23, BY-225710, Pinsk, Belarus 2Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China 3Department of Biology, National Museum of Natural Science, Taichung, Taiwan 40419, R. O. C. (Accepted: 1 Demcember 2013) ABSTRACT
Hyphodontia juniperi (Xylodon juniperi) was recorded for the first time for China in Jilin Province, Hebei Province, and in Beijing Municipality. General distribution of the species, its ecological and morphological differences from H. crustosa (Xylodon crustosus) are discussed. Bayesian reconstruction of phylogeny based on ITS1, 5.8S, and ITS2 nuclear ribosomal DNA sequences demonstrated that both H. crustosa and H. juniperi belong to a species complex, which requires richer molecular sampling for separation.
Key words: Corticiaceae s.l., geography, host range, subulate cystidia
Introduction (Pers. : Fr.) J. Erikss. [Xylodon crustosus (Pers.) Chevall.] are discussed below. Hyphodontia juniperi is a corticioid fungus, the member of Schizoporaceae Material and methods (Hymenochaetales). The name Xylodon juniperi (Bourdot & Galzin) Hjortstam & Ryvarden is Specimens of H. juniperi were examined in applied to this taxon after splitting Hyphodontia the course of a critical study of s.l., which was based on morphological Hyphodontia-like fungi from China. Reference characters (Hjortstam and Ryvarden, 2009). The specimens are deposited in herbaria TNM and species was previously reported from Europe, MSK (acronyms follow Index Herbariorum, North and South America, Africa, and two http://sweetgum.nybg.org/ih). The morphology localities in Asia (Iran and Japan). It is fairly description is based on dry basidiocarps. common in some parts of its range, like Azores Micromorphology measurements were carried and Italy, but rarely recorded in other regions, out in the medium of 3% KOH water solution; and thus it remains poorly studied in many color for hyphae and other structures was respects. In this study, three specimens of this observed in the same solution. Average spore species were found for the first time in size was calculated as arithmetical mean for 30 mainland China. Chinese specimens of H. spores, randomly selected for each specimen in juniperi and their relation to H. crustosa basidioma sections or squash preparations.
*Corresponding author, e-mail: [email protected] 46 Fung. Sci. 28(1), 2013
Spore quotient (Q) was calculated as spore additional studies of spore morphology were length/width ratio. Crystalline material carried out in this medium also. morphology was examined in Melzer’s reagent;
Table 1. Reference ITS1-5.8S-ITS2 sequences used in this research* GenBank Species Country / host Isolate / Specimen voucher accession no. Hyphodontia crustosa Germany / Fraxinus excelsior GEL 2325 DQ340313 H. crustosa Germany / Alnus incana GEL 5336 DQ340314 H. crustosa Germany / Alnus incana GEL 5360 DQ340315 H. crustosa Finland / angiosperm / KHL 11731 (GB) DQ873614 H. crustosa Taiwan / not identified Wu 0711-90 / TNM F22360 KC460312 H. crustosa China / angiosperm Wu 1007-31 / TNM F25228 KC460313 H. hastata Norway / Picea abies GEL 2143 DQ340323 (Litsch.) J. Erikss. H. juniperi China / Thuja orientalis Wu 0910-95 / TNM F23863 JX175047 H. juniperi China / angiosperm Wu 0308-26 / TNM F15237 KC460311 Réunion Island / H. juniperi Cyathea excelsa GEL 4940 DQ340316
H. pruni (Lasch) Sweden / / Ryberg 021018 (GB) DQ873624 6YUþHN H. rimosissima Sweden / / Ryberg 021031 (GB) DQ873627 (Peck) Gilb. Resinicium bicolor (Alb. & Schwein.) Lithuania / Pinus mugo Z-3-4 / FJ872065 Parmasto * Taxa in bold indicate original sequences from this study
For molecular phylogeny study, DNA was selected as the outgroup. extracted from cultural mycelium as described Sequences were aligned with MAFFT earlier in Wu et al. (2007). Living cultures are software online preserved in TNM culture collection. ITS region (http://mafft.cbrc.jp/alignment/server), using of nuclear ribosomal DNA was amplified using Q-INS-i strategy. Final hand cut of the aligned primer pair ITS1/ITS4 (White et al., 1990). sequences was done in BioEdit 7.0.4.1 software. Amplicons were sequenced as desribed in Wu et Ready data matix was deposited in TreeBASE al. (2010). Sequences were submitted to (http://purl.org/phylo/treebase/phylows/ GenBank (Tab. 1). To assess phylogenetic study/TB2:S14991). MrModeltest 2.3 by J.A.A. distances, ITS sequences of H. juniperi, H. Nylander (http://www.abc.se/~nylander/ crustosa and some other allied species with mrmodeltest2/mrmodeltest2.html) with the subulate cystidia were involved in this research. Akaike information criterion (Posada and Resinicium bicolor (Hymenochaetales) was Buckley, 2004) was used to assess the best-fit
Hyphodontia juniperi from China 47 models of nucleotide evolution for ITS1+ITS2 substratum up to 3.5 ȝP ZLGH Subhymenium and 5.8S sub-regions separately. The suggested little differentiated from subiculum; subbasidial models were implemented in partitioned hyphae moderately branched, 2.5–3 ȝP ZLGH Bayesian reconstruction of phylogeny using thin-walled, fairly richly encrusted. Hymenium MrBayes vers. 3.2.1 (Ronquist and Huelsenbeck, moderately dense. Cystidia of one type, in 2003). Eight Metropolis-coupled Markov chain hymenium, clamped at base, flexuous-subulate, Monte Carlo chains with temperature 0.2 were slightly constricted, apically attenuate, seldom initiated, and these were run for 1M generations, almost capitulate, 20–30(–50) × 2.5– ȝP in two independent runs, with tree and colorless, thin- or almost thin-walled, smooth to parameter sampling every 100 generations. The loosely encrusted, some ones projecting over the initial burn-in was set to 0.5. A majority-rule hymenium up to 15–20 ȝPBasidia clamped at consensus phylogram was computed from the base, subcylindrical to utriform, with 1–2 remaining 5000 trees. constrictions, 20–25 × 3.5– ȝP colorless, thin-walled, smooth or slightly encrusted, with Results four sterigmata 3–4 × 0.3– ȝP Linear repetition of basidia seldom. Basidiospores Three specimens of Hyphodontia juniperi ellipsoid to oblong, 5–7.5 × 2.5–3.5 ȝP rarely was identified from different localities in N and up WRȝPORQJ colorless, thin-walled, with NE China (see below in Specimens examined). one or more oily drops, with a small or unclear Morphological description of H. juniperi based apiculus. on Chinese material, is given below.
Specimens examined. China. Jilin Province, Hyphodontia juniperi (Bourdot & Galzin) J. Antu County, in the vicinity of Erhtao town, Erikss. & Hjortstam Figs. 1, 2 Shangbaishan, 42.11ºN, 128.10ºE, alt. 1,050 m, Basidioma effused, subceraceous or on dead, partly corticated twig of Abies sp. subcretaceous, continuous or minutely fissured about 1 cm diam, coll. S.H. Wu and S.Z. Chen, with age, extending from 2–10 mm to about 10 4.IX.2003, Wu 0309-130 (TNM F15343). Hebei cm, 50–120(– ȝP WKLFN in section. Province, Wulingshan, 20.36ºN, 117.29ºE, alt. Hymenophore smooth to sparsely grandinioid, 1,900 m, on twig of angiosperm species, coll. with 4–10 granules/cm. Hymenial surface pale S.H. Wu and S.Z. Chen, 29.VIII.2003, Wu cream to cream. Margin zone white, somewhat 0308-26 (TNM F15237; duplicate in MSK). thinning out and powdery, up to 0.5 mm broad. Bejing Municipality, Tanzhesi west of Beijing, Hyphal system monomitic, hyphae clamped at 39.54ºN, 116.00ºE, alt. 340 m, on dead all septa, colorless. Subiculum moderately corticated twigs about 1–1.5 cm diam of Thuja loose, rich of mostly mid-sized to coarse orientalis L., coll. S.H. Wu, 15.X.2009, Wu crystals. Subicular hyphae moderately to 0910-95, 0910-98, 0910-103 [TNM F23863 loosely branched, thin- or slightly thick-walled, (duplicate in MSK), 23865, 23868, accordingly]. (1–)1.5–2.5(–3 ȝPZLGHWKHRQHVclose to the
48 Fung. Sci. 28(1), 2013
Fig. 1. Hymenial surface and margin zone in Hyphodontia juniperi basidiomata. A. TNM F15343 (Jilin Province). B & C. TNM F23863 (Beijing). Scale bars = 1 mm.
Hyphodontia juniperi from China 49
Fig. 2. Micromorphology of Hyphodontia juniperi from China. TNM F15343 (Jilin Province). A & B. Vertical sections through basidioma. C. Subicular hyphae. D. Crystals from subiculum. E. Cystidia. F. Basidia. G. Spores in KOH solution. H. Spores in Melzer’s reagent. TNM F23863 (Beijing). I. Linear repetition in basidia. J. Spores. Scale bars: for A = 100 ȝm, for B, E, F & I = 10 ȝm, for C, D, G, H & J = 5 ȝm.
Basidiospore variation. Notable spore northernmost locality (Jilin Province). The size and shape variation was observed among smallest Q was in specimens from Beijing specimens of H. juniperi (Tab. 2). The largest vicinity. spores were observed in specimen from the
Table 2. Spore size and shape in Hyphodontia juniperi Specimen Mean spore Locality 6SRUHVL]HUDQJH ȝP Q range Mean Q ref. no. VL]HȝP (5–)5.5–6.5(–7.5) × TNM F15343 Jilin, Erhtao 5.87 × 3.20 1.5–2.3 1.84 (2.5–)3–3.5 Hebei, TNM F15237 5–6(–8.5) × 2.8–3.2(–3.5) 5.59 × 3.08 1.4–2.7 1.82 Wulingshan Beijing, TNM F23863 5–6.5(–7) × 3–3.5 5.71 × 3.32 1.4–2.1 1.73 Tanzhesi * Size for 10% of spores in brackets
Phylogenetical study. The data matrix of independent subclade of Hyphodontia juniperi ITS region was composed of 13 sequences and within crustosa-juniperi clade. Morever, 652 nucleotide positions, including introduced specimens Wu 0910-95 and GEL4940 look like gaps. Best models of evolution selected for two separate species. Within H. crustosa Bayesian reconstruction were HKY+G for phylogenetic distances are very short, but two ITS1+ITS2 and SYM+G for 5.8S. The obtained subclades can be distinguished here with Bayesian phylogram (Fig. 3) shows that H. BPP=0.91: H. crustosa from Europe (B, C) and crustosa together with H. juniperi constitute a H. crustosa from China (D). A specimen of separate cluster (B–F) apart of three other H. juniperi from Wuling Mountains (Wu Hyphodontia species with subulate cystidial 0808-26) demonstrates remarkably high elements (A). There are three sister groups in similarity in ITS sequence with H. crustosa crustosa-juniperi clade, with Bayesian posterior from Finland (B). Specimen from Wulingshan probabily (BPP) value 0.77: phylogenetically was collected on wood of angiosperm species, distant specimen of H. juniperi from Réunion which is seldom substratum for H. juniperi. (F), H. juniperi from Beijing vicinity (E), and H. However, spores in this specimen are ellipsoid, crustosa subclade (B, C, D). Thus, there is no an not cylindrical as in H. crustosa.
50 Fung. Sci. 28(1), 2013
Fig. 3. Bayesian phylogram of phylogenetic relationships of Hyphodontia species with subulate cystidia inferred from internal transcribed spacer (ITS) sequences. Numbers above branches indicate Bayesian posterior probabilities (PP > 0.75).
Discussion Honshu Island). Africa: Ethiopia (central part), According to the published records Kenya (central part), Tanzania (north). Islands (Eriksson et al., 1981; Hallenberg, 1981; of south-west Indian Ocean: Réunion (E. Gilbertson and Blackwell, 1985; Tellería, 1990; Langer, pers. comm.). North America: USA Langer, 1994; Rodríguez-Armas and Beltrán (Texas), Canada (southwest, Vancouver Island). Tejera, 1995; Melo and Tellería, 1992, 1997; South America: Colombia. Hjortstam and Ryvarden, 1997, 2004; Hyphodontia juniperi occurs in Vesterholt, 1997; Ghobad-Nejhad et al., 2009; boreonemoral, nemoral and mediterranean types Telleria et al., 2009; Gorjón et al., 2009; of biomes. The identity of the collections made Bernicchia and Gorjón, 2010; Dämon et al., from tropical regions (Africa, Canary Islands, 2010), H. juniperi has predominantly South America, Réunion) with H. juniperi temperate-holarctic distribution, as below. needs further study. As shown in its distribution, Europe: Norway, Sweden (Gotland Island), H. juniperi prefers to grow in warm and Estonia, Denmark, Germany (southwest), moderately dry regions, more often on low France (south-central part – type locality for mountains or stony areas. The findings in China Corticium serum (Pers.) Fr. var. juniperi Bordot correspond with ecogeographical pattern of the & Galzin), Austria, Portugal, Spain (north and species. One specimen from China was on central part), Italy, Croatia, Macedonia, Greece Abies, which is a new host for this species. (Crete). Macaronesia: Canary Islands, Azores. Smooth or almost smooth hymenial surface, Caucasus: Russia (Krasnodar krai, Black Sea apically tapering cystidia, and ellipsoid spores coast). Asia: Iran (north), Japan (center of are the main diagnostic features of H. juniperi.
Hyphodontia juniperi from China 51
Nevertheless, there are variations both in species as described in the key below. specimens examined by us and in the published * Hymenial surface typically whitish and diagnoses. Hymenial surface is described as smooth, not cracking into polygons; even [Jülich and Stalpers, 1980, under the name cystidia often with attached crystals, Kneiffiella juniperi (Bourdot & Galzin) Jülich never distinctly papillate or capitulate & Stalpers], or initially smooth, then apically; basidia 15–25 um long; spores subtuberculate or minutely odontioid in some ellipsoid to oblong, (2.5–)3–3.5(– ȝP areas (Rodríguez-Armas and Beltrán Tejera, broad, straight, convex or seldom barely 1995; Melo and Tellería, 1992; Eriksson et al., pressed-in adaxially; on Cupressaceae 1981; Hjortstam and Ryvarden, 2004). (Juniperus, Cupressus, Thuja, Hymenial surface in Chinese specimens of H. Cryptomeria), occasionally on other juniperi is completely smooth or very poorly gymnosperm and angiosperm genera ... H. tuberculate. juniperi Spores in H. juniperi are described as — Hymenial surface usually cream-colored, ellipsoid (Jülich and Stalpers, 1980; Eriksson et with small, distinctly separated aculei al., 1981; Melo and Tellería, 1992; Langer, when young, with age becoming 1994; Rodríguez-Armas and Beltrán Tejera, odontioid with closer, sometimes crowded 1995; Bernicchia and Gorjón, 2010), whereas aculei; old and dry specimens often Hjortstam and Ryvarden (1997; 2004) becoming cracked into small polygons; characterized spores as broadly ellipsoid. cystidia usually naked, some ones apically Spores are depicted as narrowly ellipsoid and with conidium-like segment; basidia 20– oblong in Langer (1994, Abb. 57). The Chinese 35 um long; spores narrowly ellipsoid to materials bear spores for the most part narrowly cylindrical, sometimes suballantoid, 2.5–3 ellipsoid. ȝP EURDG D SDUW RI VSRUHV SUHVVHG-in Hyphodontia juniperi belongs to H. adaxially; on angiosperms, occasionally crustosa complex, and variation in spore shape on gymnosperms ... H. crustosa makes the hiatus between H. juniperi and H. Acknowledgments crustosa somewhat diffuse. We believe that in all controversial cases an additional diagnostic The project was supported from the National criterion of H. juniperi is the ecology, namely a Science Council of Taiwan, grant no. NSC gymnosperm host and the preference of 101-2621-B-178-001-MY3. This research was moderately dry, mountain habitats. Based on financially supported by the National Science DNA sequence data we conclude that H. Council of Taiwan, grant no. NSC crustosa plus H. juniperi is a complicated 100-2811-B-178-001 for the first author; K.T. species complex, requiring more molecular Li Foundation for the Development of Science sampling for species delimitation. Until such and Technology (Taiwan) supported the study kind of research will be done, we propose the of the second author. Information on some working taxonomical concepts for the two Hyphodontia collections was kindly provided
52 Fung. Sci. 28(1), 2013 by Ewald Langer (Kassel), Karl-Henrik Larsson Aphyllophorales) from Colombia collected (Oslo), and Ivan Zmitrovich (St-Petersburg). by Leif Ryvarden. Mycotaxon 64: 229–241. Hjortstam, K. and L. Ryvarden. 2004. Some References new and noteworthy corticioid fungi Bernicchia, A. and S.P. Gorjón. 2010. (Basidiomycotina, Aphyllophorales) from Corticiaceae s. l. (Fungi Europaei), 1008 pp. Japan. Synopsis Fungorum 18: 8–13. Edizioni Candusso. Hjortstam, K. and L. Ryvarden. 2009. A Dämon, W., A. Hausknecht, and I. checklist of names in Hyphodontia sensu Krisai-Greilhuber (eds). 2010. Database of stricto – sensu lato and Schizopora with fungi in Austria. Austrian Mycological new combinations in Lagarobasidium, Society. Retrieved 24.X.2010 at Lyomyces, Kneiffiella, Schizopora, and
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Mycol. 160. 456 pp. aphyllophoroid and gastromycetoid Ronquist, F. and J.P. Huelsenbeck. 2003. basidiomycetes: 206–216. Copenhagen: MrBayes 3: Bayesian phylogenetic Nordsvamp. inference under mixed models. White, T.J., T. Bruns, S. Lee, and J.W. Taylor. Bioinformatics 19: 1572–1574. 1990. Amplification and direct sequencing Tellería, M.T. 1990. Annotated list of the of fungal ribosomal RNA genes for Corticiaceae, sensu lato (Aphyllophorales, phylogenetics. In: Innis, M.A., D.H. Basidiomycotina), for Peninsular Spain and Gelfand, J.J. Sninsky, and T.J. White (eds), Balearic Islands. Bibl. Mycol. 135. 152 pp. PCR protocols: A guide to methods and Telleria, M.T., I. Melo, M. Dueñas, I. Salcedo, J. applications. Pp. 315–322. New York: Cardoso, J.L. Rodríguez-Armas, and E. Academic Press, Inc. Beltrán-Tejera. 2009. Corticioid fungi Wu, S.H., D.M. Wang, and E. Tschen. 2007. (Basidiomycota) from the Azores Islands: Brunneocorticium pyriforme, a new Flores and São Miguel, 22 pp. Accessible at: corticioid fungal genus and species http://www.mycotaxon.com/resources/chec belonging to the euagarics clade. Mycologia klists/telleria-v109-checklist.pdf [Summary: 99: 302–309. Mycotaxon 109: 141–144]. Wu, S.H., D.M. Wang, and S.Y. Yu. 2010. Vesterholt, J. 1997. Chaetoporellaceae Jülich. In: Neoaleurodiscus fujii, a new genus and new Hansen, L. and H. Knudsen (eds), Nordic species found at the timberline in Japan. macromycetes. Vol. 3. Heterobasidioid, Mycologia 102: 217–223
㚍䰰渐劊濑Hyphodontia juniperi濒濕ᳫ◉㑮䮾懂㎒⨎劊ġ
1 Eugene Yurchenko ˣ䄲䲭曆 2ˣ⏛倚厗 3ġ
1Department of Biotechnology, Paleski State University, Dnyaprouskai flatylii str. 23, BY-225710, Pinsk, Belarus 2 ᷕ䥹昊㾳春ㅱ䓐䓇ン䞼䨞炻㾳春ⶪ
3 ⚳䩳冒䃞䥹⬠⌂䈑棐䓇䈑⬠䳬炻⎘ᷕⶪ
㐀 天
⇢㝷䴚滺却 (Hyphodontia juniperi) 椾㫉䘤䎦㕤ᷕ⚳⎱㜿䚩ˣ㱛⊿䚩⊿Ṕⶪˤ㛔㔯᷎妶婾⇢㝷䴚滺却䘬↮Ự炻 ẍ⍲⬫冯㭤䴚滺却ġ (H. crustosa) 䓇ン冯⼊ン䘬ⶖ䔘ˤ䴻䓙居叱㮷⮶↢⺷↮㜸㟠䱾橼ℏ廱抬攻昼⋨ (ITS region)ġ ⸷↿炻䳸㝄栗䣢⇢㝷䴚滺却冯㭤䴚滺却Ⱄ㕤⎴ᶨᾳ墯⎰䧖ˤ暨天㚜⣂㧁㛔䲵ℍ䞼䨞炻⥳傥⋨⇍忁ℑ䧖ˤ
斄挝娆烉䙖㭤却ˣ⛘䎮⬠ˣ⭬ᷣ䭬⚵ˣ抸䉨♲䉨橼ˤ