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AR TICLE Two New Species of Hydnoid-Fungi from India IMA FUNGUS · VOLUME 4 · NO 2: 359–369 I#JKK$'LNJ#*JPJN#K Two new species of hydnoid-fungi from India ARTICLE Kanad Das1, Joost A. Stalpers2, and J. Benjamin Stielow2 1Botanical Survey of India, Cryptogamy unit, Howrah, 711103, India 2/x"?|;<x/~x6'K#|+?*KJ';_@|>?I knaw.nl Abstract: @6Hericium yumthangense (Russulales, Agaricomycotina) and Mycoleptodonoides sharmae Key words: (Polyporales, Agaricomycotina) are described as new to science from the Shingba Rhododendron sanctuary Abies located in the northern district of Sikkim, India. Macro- and micromorphological characters are described and Hericiaceae 6@"|; H. yumthangense Himalaya as a rather isolated species within Hericium 6 Meruliaceae [ M. sharmae N'" |; #LN Rhododendron ?6? Sikkim 6 Article info:"I¤NJ#*>;INNJ#*>~I#*NJ#* INTRODUCTION 6 G NJJK> OP < Covering an area of 43 sq. km, the Shingba Rhododendron were taken with a Nikon D300s camera. sanctuary is located in the Yumthang valley of the northern Micromorphological features were studied from dried district of Sikkim, India, in eastern Himalaya, surrounded by 6K#6/ China, Bhutan, and Nepal. It is characterized by temperate red and 30% glycerol, and Melzer’s reagent. Drawings were [ Rhododendron forest and is a native habitat made with a drawing tube (attached to an Olympus CX41 for Rhododendron niveum. Apart from the trees like Abies #JJJ&x6? densa, 8/[, Picea spinulosa, Magnolia globosa, ?6 M. campbellii, Acer pectinatum, Betula utilis, and more than ornamentations (if any). Measurements are based on 20 40 species of Rhododendron, this natural area is also rich 6 x \ given as KDa-KDc?&6?KDz-KDy in which KDa is the including Primula, Potentilla, Ranunculus, Euphorbia, 6 Roscoea, Heracleum, Rubus, and Aconitum. the length, KDc6 During a macrofungal survey of the Shingba Rhododen- 6 dron sanctuary and adjacent areas by K.D. between August for the width, and KDz the mean value for the width. The and September 2011, various saprotrophic wood-decaying ?£+L and mycorrhizal macromycetes were collected. Macro- and ?? micromorphological studies revealed some of them were 6 6@ measured collections, and the mean quotient value. here as Hericium yumthangense sp. nov. (Hericiaceae) and Scanning electron micrographs of basidiospores were Mycoleptodonoides sharmae sp. nov. (Meruliaceae). Macro-, obtained from dry spore prints directly mounted on double- micro- and ultrastructural illustrations, together with detailed sided adhesive tape, pasted on a metallic specimen-stub, |;@"N'" gold coated, and subsequently scanned in high vacuum mode and mtSSU DNA barcodes are provided. [? @<R 200 model scanning electron microscope (SEM) at the S.N. MATERIALS AND METHODS Bose National Centre for Basic Sciences, Kolkata (India). Morphological studies DNA isolation, PCR, and sequencing Macromorphological characters were observed and recorded @|;6 [/ ¤ |; ` follow the chart in the Flora of British Fungi ;#$$>OP @~/[ © 2013 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: [ Non-commercial: No derivative works: For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author’s moral rights. VOLUME 4 · NO. 2 359 Das et al. Creolophus cirrhatus voucher RGB Kew K(M) 135658 (EU784261) 79 Hericium corraloides (Europe) Hericium coralloides s.l. 91 Hericium corraloides (China/North America) Hericium erinaceus #1 (Korea/China/North America) Hericium erinaceum strain KUMC e1 (AY534596) Hericium erinaceum CBS 233.87 (DQ185921) ARTICLE Hericium erinaceum CBS 485.95 (AY534583) Hericium erinaceus #2 (North America) Hericium erinaceum ATCC 56457 (DQ185920) Hericium erinaceum CBS 260.74 (DQ185922) 75 Hericium erinaceum voucher RBG Kew KM62494 (EU784265) 100 Hericium coralloides (EU784265) Hericium coralloides (AF173216) Hericium coralloides IFO 7716 (AY534582) Hericium americanum Hericium americanum CBS 493.63 (AY534581) Hericium erinaceum CBS 204.76 (DQ185924) Hericium americanum (AF287980) Hericium yumthangense KD-11-146 (JX855033) Hericium americanum AFTOL-ID 469 (DQ206987) Hericium cf. alpestre KM 107270 RBG Kew (EU784259) 78 Hericium abietis (AF287979) Hericium abietis VC 63-12 (JQ716938) Hericium abietis 77 Hericium abietis CBS 243.48 (AY534579) Hericium abietis HMJAU20029 (JF430067) Hericium erinaceum CBS 444.85 (DQ185927) Hericium erinaceum (AM981220) 86 Hericium alpestre FCUG 2754 (JQ716930) Hericium alpestre FCUG 2754 (JQ716930) Hericium alpestre Hericium erinaceum (AM269779) Hericium erinaceum (AM981226) Hericium erinaceum CBS 449.85 (DQ185928) Hericium erinaceum KUMC 1035 (DQ185919) 100 Hericium bharengense KD-10658 (JX855036) Hericium bharengense KD-10658E (JN185603) Laxitextum bicolor (AM269787) 0.04 Fig. 1. ~6?4+@"|;Hericium dataset. Numbers #JJJJ6 +`x @"#?K'"?@"N |; @"KL nucleotide sequence database collaboration (INSDC) ITS1 and ITS4 was performed under standard conditions (White 6[ et al. 1990, Stielow et al. 2010). PCR conditions for amplifying alignment ambiguities, and the resulting phylogenies. Since ""_ N'" |; almost no sequence data is yet available for the genus MS1, MS2, LR0R and LR5 only differed in their annealing Mycoleptodonoides (INSDC contained a single ITS sequence KK / J / [ 6$JL~/[ N'" #LN using FastAP thermosensitive alkaline phosphatase and query were retrieved viax+;"@[ @ " @ ? respect to the quality criteria described above, and the dataset sequencing reaction was set up using ABI big dye terminator v. was further supplemented with sequences chosen according 3.1, using a quarter of the suggested volumes (manufacturer's to Binder et al. (2005). Sequences for both datasets were protocol), followed by bidirectional sequencing with a laboratory subsequently aligned, using the EMBL-EBI MAFFT server capillary electrophoresis system (Lifetechnologies, 3730XL ILLL@LLL DNA analyzer). Sequences were stored and manually corrected ; `?` et al. for sequencing artifacts and forward and reverse sequences NJ## 6? assembled using the Biolomics database (www.bio-aware. 4+<#$'#;64+ " |/x` N `@?4 O@6P approach (Stamatakis et al.NJJ'46? for the Hericium dataset were rooted with Laxitextum bicolor, Phylogenetic inference chosen as outgroup according to Larsson et al. (2004). The A search for respective Hericium@" Æ#JJ phlebioid outgroup (Phlebia spp.) for the Mycoleptodonoides sequences, which were downloaded from the international dataset and clade names were depicted according to Binder 360 IMA FUNGUS Hydnoid fungi from eastern Himalaya 83 Antrodia malicola BCRC 35452 (AY333837) Antrodia malicola MJL 1167SP (AY333835) ARTICLE Fomitopsis rosea FP 104278T (AY333809) 83 100 89 Fomitopsis rosea RLG 6954T (AY333808) Fomitopsis cajanderi BCRC 35447 (EU232294) Antrodia serialis FP 105599 (EU232293) 78 Antrodia juniperina FP 97452T (AY333839) 71 Antrodia juniperina (AY333838) 99 Antrodia juniperina (AY333839) Antrodia clade #1 82 100 Antrodia sp. TFRI 782 (EU232276) Antrodia sp. TFRI 781 (EU232275) 73 Fomitopsis pinicola FP 98533T (AY333810) 89 Fomitopsis pinicola AFTOLID 770 (AY684164) Fomitopsis pinicola FP 105760T (AY333811) 85 Fomitopsis pinicola TFRI 513 (EU232290) Fomitopsis pinicola (AF287858) 100 Antrodia heteromorpha FCUG 1244 (AY333840) Antrodia albida FCUG 1396 (AY333845) Radulomyces notabilis EL 5/97 (EU118664) Cinereomyces lindbladii Larsson 12078 97 Diplomitoporus crustulinus FP 101824 (AY333816) 100 Diplomitoporus lindbladii HHB 5629 (AY333814) 99 Diplomitoporus lindbladii KEW212 (AF518615) Gelatoporia subvermispora Kotiranta 20823 (FN907911) 100 96 100 Obba valdiviana FF503 (HQ659235) Obba valdiviana Rajchenberg 336 (HQ659236) Mycoleptodonoides sharmae KD-11-122 (JX855030) Diplomitoporus overholtsii HHB 9465 (AY333813) 100 Pycnoporus puniceus BCC27595 (FJ372708) 78 Pycnoporus sanguineus 8R12 (FJ372694) Core polyporoid clade Dichomitus squalens (HQ659224) 92 Perenniporia medulla-panis (JN112793) 79 Ganoderma lucidum (AB368068) 76 Polyporus tricholoma 9591 (AB368100) 77 Polyporus umbellatus WD719 (AB368109) 97 100 Polyporus squamosus MUCL30721 (AB368094) Polyporus squamosus WD2380 (AB587646) Dentocorticium ussuricum CBS 849.84 (KF691613) 100 Grifola frondosa ACCC 50289 (AY826982) Grifola frondosa AFTOL ID 701 (AY629318) 100 Antrodia carbonica FP 105585R (EU232285) Antrodia carbonica (AF287844) Antrodia xantha TFRI 879 (EU232284) Antrodia sitchensis HHB 12513 (AY333830) 100 Antrodia xantha (AJ583430) Antrodia sp. CBS 101451 (DQ457649) Antrodia clade #2 85 Antrodia sinuosa RLG 2538 (EU232288) 80 Antrodia sitchensis HHB 5320 (EU232287) 100 Antrodia sinuosa L6192SP (AY333832) Antrodia sinuosa RLG 1182R (AY333831) 89 Antrodia odora HHB 6094 (EU232286) 100 Phlebia rufa CBS 213.47 (KF691614) Phlebia acerina CBS 125860 (KF691615) 0.03 Fig. 2. ~6?4++"_N'"|;Mycoleptodonoides dataset. Numbers on the branches represent support values from 1000 bootstrap replicates if at least 70%, with branches scaled in terms of the 6@`x et al. (2005). For depicting the trees, clades comprising at subtree with H. erinaceus, H. americanum, H. alpestre, and least twenty sequences were collapsed if they were either H. abietis, received strong bootstrap branch support (BS) 6 ' H. coralloides subtree was not
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