1208 fungal strains isolated from 120 samples of four glaciers and two snow mountains were preliminarily identified based on BLAST comparison of ITS sequences against the GenBank database. As one of the most commonly encountered fungal groups, 41 isolates belonging to Cadophora were studied in detail.
Phylogenetic analyses
Sequences of referential species, especially those of ex-type strains, were retrieved from GenBank and added to the sequences generated in this study (Table 2). The alignments of partial sequences of LSU, ITS, LSU, TUB and TEF1-α have 855, 446, 828, 567, 693 characters respectively.
Table 2
Strains analyzed in this study, with collection details and GenBank accession numbers
Species | Strain no. | Host/ substrate | Country | GenBank Accession No. |
LSU | ITS | TUB | TEF1-α |
Articulospora tetracladia | DSM 104345 | – | – | MK226456 | MH930816 | MK241460 | MK241447 |
Ascocorticium anomalum | CBS 874.71 | – | Germany | MH872135 | – | – | – |
Cadophora anfricana | CBS 120890T | Prunus salicina, necrotic wood | South Africa | MT156170 | MN232936 | MN232967 | MN232988 |
Cadophora antarctica | FMR16056T | diesel-contaminated soil sample | Antarctica | MG385663 | MG385664 | – | – |
Cadophora bubakii | CBS 198.30T | margarine | Czech Republic | MH866559 | MH855111 | – | MN232989 |
Cadophora caespitosa | CGMCC3.20179 = MY156T | water in Mingyong Glacier | China | MT908194 | MT889936 | MT921201 | MT900568 |
Cadophora caespitosa | CGMCC3.20180 = MY169 | water in Mingyong Glacier | China | MT908195 | MT889937 | MT921202 | MT921172 |
Cadophora caespitosa | CGMCC3.20192 = DG1120 | water in Dagu Glacier | China | MT908222 | MT889964 | MT921229 | MT921197 |
Cadophora caespitosa | CGMCC3.20431 = HL674 | water in Hailuogou Glacier | China | MW793546 | MW793520 | MW818434 | MW810619 |
Cadophora caespitosa | CGMCC3.20432 = BM691 | soil in Baima Snow Mountain | China | MW793547 | MW793521 | MW818435 | MW810620 |
Cadophora constrictospora | P1751T | endophytic in roots of Microthlaspi | Bulgaria | MN339369 | KT269023 | – | MN325874 |
Cadophora dextrinospora | AG5 | decayed wood in Anoplophora glabripennis galleries | Finland | – | MF188986 | – | – |
Cadophora dextrinospora | CBS 401.78T | decaying wood | Spain | MH872917 | NR_119489 | – | – |
Cadophora echinata | P6045T | endophytic in roots of Microthlaspi perfoliatum | Spain | MN339428 | KT270239 | – | MN325932 |
Cadophora fallopiae | CPC 35742 | Reynoutria japonica | Germany | MT223877 | MT223782 | – | – |
Cadophora fascicularis | P2794T | endophytic in roots of Microthlaspi erraticum | Germany | MN339414 | KT269992 | – | MN325918 |
Cadophora fastigiata | CBS 307.49 | Pine wood | Sweden | MH868062 | MH856538 | KM497131 | KM497087 |
Cadophora ferruginea | P1323T | endophytic in roots of Microthlaspi perfoliatum | Spain | MN339356 | KT268618 | – | MN325861 |
Cadophora gamsii | P2437T | endophytic in roots of Microthlaspi erraticum | France | – | KT269668 | – | MN325899 |
Cadophora gregata | ATCC 11073T | Glycine max, brown stem rot | Japan | MF979571 | U66731 | MF677920 | MF979586 |
Cadophora helianthi | CBS 144752T | Helianthus annuus, necrotic tissue in stem | Ukraine | – | MK813837 | MH733391 | MH719029 |
Cadophora indistincta | CGMCC3.20233 = DG978 | soil in Dagu Glacier | China | MT908210 | MT889952 | MT921217 | MT921186 |
Cadophora indistincta | CGMCC3.20234 = DG1054 | water in Dagu Glacier | China | MT908215 | MT889957 | MT921222 | MT921191 |
Cadophora indistincta | CGMCC3.20189 = DG1014T | water in Dagu Glacier | China | MT908211 | MT889953 | MT921218 | MT921187 |
Cadophora indistincta | CGMCC3.20195 = DG1017 | soil in Dagu Glacier | China | MT908212 | MT889954 | MT921219 | MT921188 |
Cadophora indistincta | CGMCC3.20196 = DG1074 | soil in Dagu Glacier | China | MT908219 | MT889961 | MT921226 | MT921194 |
Cadophora inflata | CGMCC3.20186 = MY759T | soil in Mingyong Glacier | China | MT908204 | MT889946 | MT921211 | MT921181 |
Cadophora interclivum | CBS143323 = BAG4T | Carex sprengelii, root | Canada | MF979565 | MF979577 | MF677917 | MF979583 |
Cadophora lacrimiformis | MFLU 16-1486T | unknown Brassicaceae, dead stem | Russia | MK591959 | MK585003 | – | – |
Cadophora luteo-olivacea | CBS 141.41T | waste water | Sweden | MH867586 | MH856092 | KM497133 | JN808856 |
Cadophora luteo-olivacea | GLMC 517 | Prunus domestica, necrotic wood | Germany | – | MN232937 | MN232968 | MN233003 |
Cadophora magna | CGMCC3.20188 = MY902T | soil in Mingyong Glacier | China | MT908208 | MT889950 | MT921215 | MT921184 |
Cadophora malorum | CBS 165.42 | Amblystoma mexicanum | Netherlands | MH867607 | MH856109 | KM497134 | KM497090 |
Cadophora malorum | CGMCC3.20184 = YL412 | soil in Yulong Snow Mountain | China | MT908200 | MT889942 | MT921207 | MT921177 |
Cadophora margaritata | CBS 144084 | Colonized wood | Finland | – | MH203866 | – | – |
Cadophora margaritata | CBS144083T | Colonized wood | Finland | MH267288 | KJ702027 | MH327786 | – |
Cadophora melinii | CBS 268.33T | probably wood-pulp | Sweden | MH866887 | NR_111150 | KM497132 | KM497088 |
Cadophora melinii | ONC1 | Vitis vinifera 'Cabernet Franc', wood canker | Canada | – | KM497033 | KM497114 | KM497070 |
Cadophora melinii | U11 | Vitis vinifera 'Sangiovese', vascular discoloration | USA | – | KM497032 | KM497113 | KM497069 |
Cadophora meredithiae | CBS143322 = BAG2T | Carex sprengelii, root | Canada | MF979568 | MF979574 | MF677914 | MF979580 |
Cadophora neoregeliae | CBS 146821T | from leaf spots of Neoregelia sp. | New Zealand | MZ064468 | MZ064411 | – | – |
Cadophora novi-eboraci | GLMC 239 | Prunus cerasus, necrotic wood | Germany | – | MN232942 | MN232973 | MN232990 |
Cadophora novi-eboraci | GLMC 273 | Prunus cerasus, necrotic wood | Germany | MT156177 | MN232943 | MN232974 | MN232991 |
Cadophora novi-eboraci | NYC14T | Vitis labruscana, wood canker | USA | – | KM497037 | KM497118 | KM497074 |
Cadophora novi-eboraci | CGMCC3.20190 = YZ1034 | soil in Yanzigou Glacier | China | MT908213 | MT889955 | MT921220 | MT921189 |
Cadophora novi-eboraci | CGMCC3.20434 = YZ1026 | soil in Yanzigou Glacier | China | MW793552 | MW793526 | MW818436 | MW810622 |
Cadophora obovata | P1963T | endophytic in roots of Microthlaspi erraticum | Germany | MN339384 | KT269230 | – | MN325888 |
Cadophora orchidicola | UAMH 8152 | Pedicularis bracteosa, root | Canada | MF979572 | AF214576 | MF677921 | MF979587 |
Cadophora orientoamericana | CTC5 | Vitis hybrid 'Cayuga white', wood canker | USA | – | KM497015 | KM497096 | KM497052 |
Cadophora orientoamericana | MYA-4972 = NHC1T | Vitis vinifera ‘Niagara’ | USA | MF979573 | KM497018 | KM497099 | KM497055 |
Cadophora prunicola | CBS 120891T | Prunus salicina, necrotic wood | South Africa | MT156182 | MN232949 | MN232979 | MN232997 |
Cadophora prunicola | GLMC 276 | Prunus cerasus, necrotic wood | Germany | – | MN232951 | MN232980 | MN232998 |
Cadophora psychrophila | CGMCC3.20845 = DG5 | soil in Dagu Glacier | China | OL477357 | OL477351 | OL674144 | OL674147 |
Cadophora psychrophila | CGMCC3.20846 = DG21T | soil in Dagu Glacier | China | OL477356 | OL714365 | OL674143 | OL674146 |
Cadophora qinghai-tibetana | CGMCC3.20181 = BM327 | soil in Baima Snow Mountain | China | MT908197 | MT889939 | MT921204 | MT921174 |
Cadophora qinghai-tibetana | CGMCC3.20182 = YL357 | water in Yulong Snow Mountain | China | MT908198 | MT889940 | MT921205 | MT921175 |
Cadophora qinghai-tibetana | CGMCC3.20183 = BM360 | soil in Baima Snow Mountain | China | MT908199 | MT889941 | MT921206 | MT921176 |
Cadophora qinghai-tibetana | CGMCC3.20185 = MY474 | soil in Mingyong Glacier | China | MT908202 | MT889944 | MT921209 | MT921179 |
Cadophora qinghai-tibetana | CGMCC3.20191 = DG1048 | soil in Dagu Glacier | China | MT908214 | MT889956 | MT921221 | MT921190 |
Cadophora qinghai-tibetana | CGMCC3.20193 = DG1156T | soil in Dagu Glacier | China | MT908223 | MT889965 | MT921230 | MT921198 |
Cadophora qinghai-tibetana | CGMCC3.20194 = YL414 | water in Yulong Snow Mountain | China | MT908201 | MT889943 | MT921208 | MT921178 |
Cadophora qinghai-tibetana | CGMCC3.20197 = DG1105 | soil in Dagu Glacier | China | MT908221 | MT889963 | MT921228 | MT921196 |
Cadophora qinghai-tibetana | CGMCC3.20228 = YL73 | soil in Yulong Snow Mountain | China | MT908193 | MT889905 | MT921200 | MT898424 |
Cadophora qinghai-tibetana | CGMCC3.20229 = YL319 | water in Yulong Snow Mountain | China | MT908196 | MT889938 | MT921203 | MT921173 |
Cadophora qinghai-tibetana | CGMCC3.20230 = BM523 | soil in Baima Snow Mountain | China | MT908203 | MT889945 | MT921210 | MT921180 |
Cadophora qinghai-tibetana | CGMCC3.20231 = MY873 | soil in Mingyong Glacier | China | MT908207 | MT889949 | MT921214 | MT921183 |
Cadophora qinghai-tibetana | CGMCC3.20232 = DG975 | soil in Dagu Glacier | China | MT908209 | MT889951 | MT921216 | MT921185 |
Cadophora qinghai-tibetana | CGMCC3.20235 = DG1073 | soil in Dagu Glacier | China | MT908218 | MT889960 | MT921225 | MT921193 |
Cadophora qinghai-tibetana | CGMCC3.20236 = DG1087 | soil in Dagu Glacier | China | MT908220 | MT889962 | MT921227 | MT921195 |
Cadophora qinghai-tibetana | CGMCC3.20433 = BM857 | soil in Baima Snow Mountain | China | MW793551 | MW793525 | MW818439 | MW810621 |
Cadophora qinghai-tibetana | CGMCC3.20435 = YL305 | water in Yulong Snow Mountain | China | MW793548 | MW793522 | MW818433 | – |
Cadophora qinghai-tibetana | CGMCC3.20436 = BM816 | soil in Baima Snow Mountain | China | MW793550 | MW793524 | MW818438 | – |
Cadophora qinghai-tibetana | CGMCC3.20437 = HL876 | soil in Hailuogou Glacier | China | MW793549 | MW793523 | MW818437 | – |
Cadophora qinghai-tibetana | CGMCC3.20847 = MY492 | soil in Mingyong Glacier | China | OL477358 | OL477352 | OL674145 | OL674148 |
Cadophora qinghai-tibetana | CGMCC3.20848 = MY527 | soil in Mingyong Glacier | China | OL815016 | OL815013 | OL790381 | OL790384 |
Cadophora qinghai-tibetana | CGMCC3.20849 = MY588 | soil in Mingyong Glacier | China | OL815017 | OL815014 | OL790382 | OL790385 |
Cadophora qinghai-tibetana | CGMCC3.20850 = MY589 | soil in Mingyong Glacier | China | OL815018 | OL815015 | OL790383 | OL790386 |
Cadophora ramosa | CBS 111743 | Actinidia chinensis, vascular discoloration | Italy | – | DQ404351 | KM497136 | KM497091 |
Cadophora ramosa | GLMC 377T | Prunus cerasus, necrotic wood | Germany | MT156187 | MN232956 | MN232984 | MN233002 |
Cadophora sabaouae | WAMC117 | Vitis vinifera | Algeria | – | MT524745 | MT646750 | MT646747 |
Cadophora sabaouae | WAMC118 | Vitis vinifera | Algeria | – | MT524744 | MT646751 | MT646748 |
Cadophora sabaouae | WAMC34T | Vitis vinifera | Algeria | – | MT644187 | MT646749 | MT646746 |
Cadophora variabilis | P1176T | endophytic in roots of Microthlaspi perfoliatum | Croatia | MK539845 | KT268493 | – | MK550890 |
Cadophora viticola | Cme-1 | Vitis vinifera 'Syrah', black streaks in shoots | Spain | – | HQ661096 | HQ661096 | HQ661081 |
Cadophora viticola | Cme-2T | Vitis vinifera 'Syrah', black streaks in shoots | Spain | – | HQ661097 | HQ661097 | HQ661082 |
Cadophora yulongensis | CGMCC3.20187 = YL814T | soil sample in Yulong Snow Mountain | China | MT908206 | MT889948 | MT921213 | MT921182 |
Calycina alstrupii | Pz162T | on Lobaria pulmonaria growing on trunk of Alnus incana | Norway | KY305097 | – | – | – |
Calycina marina | TROM F26093 | dead seaweed (Ascophyllum nodosum) | Norway | KT185670 | – | – | – |
Cenangium acuum | TAAM 198449 | Pinus sylvestris | Czech Republic | KX090828 | – | – | – |
Cenangium ferruginosum | CBS 556.70 | – | Netherlands | MH871625 | – | – | – |
Cephalosporium gramineum | CBS 132.34T | Triticum aestivum, culm | Japan | NG_070839 | NR_171209 | – | – |
Chaetomella acutiseta | AFTOL-ID 270 | – | – | AY544679 | – | – | – |
Chaetomella oblonga | CBS 110.78 | leaf of Acer sp. | Canada | MH872875 | – | – | – |
Chlorociboria aeruginosa | CBS 139.28 | – | – | MH877688 | – | – | – |
Chlorociboria clavula | D1611 | – | New Zealand | JN939941 | – | – | – |
Collembolispora aristata | CPC21145T | foam in an unnamed right tributary of the brook Bezenek | Czech Republic | KC005811 | NR_111830 | – | KC005818 |
Collembolispora barbata | CBS 115944 = UMB-088.01T | mountain freshwater stream | Portugal | | NR_111443 | – | – |
Cordierites frondosa | HKAS41508 | – | – | AY789354 | – | – | – |
Cordierites guianensis | 192 | – | – | EU107270 | – | – | – |
Cudoniella clavus | AFTOL-ID 166 | – | – | DQ470944 | – | – | – |
Dermea bicolor | CBS 135.46 | – | Canada | MH867659 | – | – | – |
Dermea cerasi | CBS 432.67 | – | – | MH870721 | – | – | – |
Graphium rubrum | CBS 210.34T | – | USA | MH866974 | – | – | – |
Helgardia anguioides | CBS 496.80T | – | Germany | MH873055 | – | – | – |
Helgardia anguioides | RAN45 | – | Germany | – | AY266144 | – | – |
Hyaloscypha finlandica | CBS 444.86T | Pinus sylvestris, root of seedling | Finland | MH873675 | NR_121279 | KM497130 | KM497086 |
Hyaloscypha melinii | CBS 143705T | – | Czech Republic | NG_068558 | – | – | – |
Hyaloscypha vitreola | CBS 126276 | – | Finland | MH875413 | – | – | – |
Lachnum carneolum | CBS 231.54 | – | France | MH868838 | – | – | – |
Lachnum diminutum | CBS 232.54 | – | France | MH868839 | – | – | – |
Leotia lubrica | KKM 427 | mycorrhizal root tip | Costa Rica | KF836631 | – | – | – |
Mastigosporium album | CPC 22945T | Alopecurus pratensis | Netherlands | KJ710451 | KJ710476 | – | – |
Mastigosporium kitzebergense | CBS 270.69T | – | Germany | MH871040 | MH859306 | – | – |
Mollisia cinerea | CBS 122029 | fallen log | USA | MT026558 | – | – | – |
Mollisia cinerella | CBS 312.61 | – | France | MH869631 | MH858062 | – | – |
Mollisia discolor | CBS 289.59 | – | France | MT026504 | – | – | – |
Mollisia fallens | CBS 221.56 | – | Netherlands | MT026505 | – | – | – |
Mycochaetophora gentianae | MAFF 239231T | – | Japan | AB496937 | NR_121201 | – | – |
Mycochaetophora sp. | MAFF 239284 | – | Japan | AB469680 | AB469681 | – | – |
Neospermospora avenae | CBS 227.38T | Avena sativa | USA | NG_077377 | MW298276 | – | – |
Oculimacula acuformis | CBS 495.80T | culm base | Germany | MH873054 | MH861289 | – | MG934497 |
Oculimacula aestiva | CBS 114730 | – | Sweden | – | MG934454 | – | MG934496 |
Oculimacula yallundae | CBS 128.31 | – | France | – | MH855154 | – | MG934499 |
Oculimacula yallundae | CBS 494.80 | culm base | Germany | – | JF412009 | – | MG934500 |
Phialocephala dimorphospora | CBS 976.72 | – | Germany | MH878299 | – | – | – |
Phialophora dancoi | CBS 329.90T | – | Argentina | MH873899 | MH862214 | – | – |
Pleuroascus nicholsonii | CBS 345.73T | the dung of pack rat | USA | MH872404 | – | – | – |
Porodiplodia livistonae | CPC 32154T | Livistona australis | Australia | NG_069575 | – | – | – |
Porodiplodia vitis | CBS 144634T | Vitis vinifera | USA | MK442552 | – | – | – |
Rhexocercosporidium camporesii | MFLU 17-1594T | dead stems | Italy | MN688632 | MN688634 | – | – |
Rhexocercosporidium carotae | CBS 418.65T | – | Norway | MH870289 | NR_111086 | – | – |
Rhexocercosporidium microsporum | MFLU 18-2672T | unknown Apiaceae, stem | UK | MK591966 | MK584939 | – | – |
Rhynchosporium agropyri | H11 | – | – | – | HM627478 | – | HM627463 |
Rhynchosporium commune | H7 | – | – | – | – | HM627434 | HM627459 |
Rhynchosporium commune | H10 | – | – | – | – | HM627437 | HM627462 |
Rhynchosporium orthosporum | 04CH-Bar-A.1.1.3 | Dactylis glomerata | Switzerland | KU844335 | – | – | – |
Rhynchosporium secalis | 02CH4-6a.1 | – | Switzerland | – | KU844333 | – | – |
Rutstroemia bulgarioides | TAAM 198322 | fallen cone | Estonia | KX090836 | – | – | – |
Rutstroemia firma | CBS 115.86T | – | Netherlands | MH873619 | – | – | – |
Sclerotinia bulborum | CBS 297.31 | – | USA | MH866668 | – | – | – |
Sclerotinia sclerotiorum | WZ0067 | – | China | AY789347 | – | – | – |
Xylaria hypoxylon | CBS 120.16 | – | – | MH866173 | – | – | – |
Ypsilina buttingtonensis | CPC 39109T | from heartwood of 1000-yr-old Quercus sp. | UK | MT373355 | MT373372 | – | – |
Ypsilina graminea | CBS 114630T | – | UK | MH874529 | NR_160217 | – | – |
Tex-type strain; 1LSU: large subunit nrDNA; ITS: Internal transcribed spacers 1 and 2 together with 5.8S nrDNA; TUB: partial beta-tubulin gene; TEF1-α: partial translation elongation factor 1-alpha gene. |
According to the LSU phylogenetic tree, representative Cadophora strains of this study (marked with bold font) and the known Cadophora species were interspersed with species of other genera in Ploettnerulaceae and formed a well-supported clade (BP/BP/PP = 90/98/100, ML/MP bootstrap and BI posterior probability support values, respectively) that distinctly separated from other family members in the Helotiales (Fig. 3).
A multi-gene phylogenetic tree was also employed to investigate further phylogenetic relationships intra and among Cadophora and allied genera (Fig. 4). All the representative species clustered into two main clades with high ML/MP bootstrap or BI posterior probability support values (92/80/100, 97/-/100 respectively). In the first main clade (Clade1), 38 isolates of this study formed six distinct subclades: isolates of YZ1026 and YZ1034 clustered in a lineage including the ex-type sequences of C. novi-eboraci with strong branch support; although strain MY902 and the known species of Cephalosporium gremineum formed a well supported subclade, they were obviously distinguished morphologically and the placement of C. gremineum should also be confirmed by protein coding genes which were unavailable currently; the other four subclades grouped separately with previously described species. Combined with morphological characteristics, we proposed five Cadophora species new to science: Cadophora caespitosa, C. indistincta, C. magna, C. psychrophila and C. qinghai-tibetana. Clade 1 also included most of the phialidic Cadophora species (including the type) and three species (Cephalosporium gramineum, Mollisia cinerella and Phialophora dancoi) belonged to other genera. The second main clade (Clade 2) contained the rest Cadophora species and most other Ploettnerulaceae members. Three isolates of this study were included in this clade: strain YL412 clustered with C. malorum in a well supported lineage; strain MY759 and MY814 formed two distinct single strain clades and we proposed them as two new species (Cadophora inflata and Cadophora yulongensis). Cadophora species in Clade 2 had multiform conidiogenesis modes and formed lineages interspersed by other Ploettnerulaceae members.
Taxonomy
Cadophora caespitosa Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837889.
Figure 5
Etymology
Referring to multiple phialides arranged in terminal fascicles.
Type: China: Yunnan Province, Mingyong Glacier, N28°27'25" E98°45'51", 2960 m, from water, 9 May 2017, M-M Wang, holotype HBU20001, culture ex-type MY156 = CGMCC3. 20179.
Mycelium hyaline to brown, septate, smooth-walled, branched, 1–3 µm wide. Conidiophores pale brown or hyaline, straight, septate, smooth, branched or unbranched, bearing solitary or multiple phialides. Conidiogenous cells phialidic, located laterally on fertile hyphae or arranged in complex heads, cylindrical to navicular, often constricted at the base, upper subulate, hyaline, smooth-walled, 6.49–32.32 × 2.56–3.83 µm, collarettes distinct, funnel-shaped, 1.92–3.89 µm long, opening 1.85–3.36 µm wide. Conidia hyaline, aseptate, smooth-walled, sporulation abundant, ovate to dacryoid or ellipsoidal, single, with both ends rounded, straight, 3.37–7.05 × 1.71–3.41 µm (mean = 5.02 ± 0.85 × 2.58 ± 0.41 µm, n = 30), L/W ratio = 1.95.
Culture characteristics — Colonies on MEA reaching 33 mm diam after 14 d at 25°C in the dark, on OA and PDA reaching 55 mm and 34 mm diam, respectively. Colonies on MEA with a smooth margin, flat, grey-white, buff to light yellow at the margin, reverse olive-black. Colonies on OA with an entire margin, flat, greenish-black with a white margin, reverse same colours. Colonies on PDA with an entire margin, flat, hazel to yellow-brown with a beige margin, reverse same colours.
Additional specimens examined: China: Sichuan Province, Dagu Glacier, N32°14'23" E 102°47'7", 3610 m, from water, 1 May 2017, M-M Wang, culture DG1120 = CGMCC3.20192; Sichuan Province, Hailuogou Glacier, N29°33'10" E101°58'10", 3180 m, from water, 28 Apr. 2017, M-M Wang, culture HL674 = CGMCC3.20431; Yunnan Province, Baima Snow Mountain, N28°23'29" E98°59'22", 4124.7 m, from soil, 10 May 2017, M-M Wang, BM691 = CGMCC3.20432; Yunnan Province, Mingyong Glacier, N28°27'25" E98°45'51", 2960 m, from water, 9 May 2017, M-M Wang, culture MY169 = CGMCC3.20180.
Notes — According to Day et al. (2012), the genera Cadophora and Phialocephala are generally distinguished by phialide complexity and conidial length, with the former producing solitary phialides and conidia longer than 4 µm, while the latter producing densely packed heads of phialides and conidia shorter than 4 µm. The newly described species is characterized by having distinct, dark stipe with multiple phialides terminating in a complexly penicillately branched apex. Phylogenetic analyses based on sequences of LSU and combined ITS + LSU + TUB + TEF1-α regions clearly show that C. caespitosa grouped with species of Cadophora in the family of Ploettnerulaceae and formed a well supported lineage.
Cadophora indistincta Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837895.
Figure 6
Etymology
Referring to the indistinct collarettes of phialides.
Type: China: Sichuan Province, Dagu Glacier, N32°8'19" E102°56'13", 2380 m, from water, 1 May 2017, M-M Wang, holotype HBU20012, culture ex-type DG1014 = CGMCC3.20189.
Mycelium hyaline, septate, smooth-walled, branched, 1–4 µm. Conidiophores hyaline, septate, smooth, often solitary. Conidiogenous cells phialidic, located terminally or laterally, discrete, hyaline, smooth-walled, straight or curved, cylindrical to navicular, often inflated in the middle and constricted at the base, 5.28–31.43 × 1.57–3.69 µm, collarettes indistinct, most phialides lack collarettes. Conidia hyaline, aseptate, smooth-walled, cylindrical to oblong, 4.67–7.50 × 1.62–2.54 µm (mean = 5.46 ± 0.66 × 2.17 ± 0.23 µm, n = 30), L/W ratio = 2.52.
Culture characteristics — Colonies on MEA reaching 45 mm diam, after 14 d at 25°C in the dark, on OA and PDA reaching 49 mm and 44 mm diam, respectively. Colonies on MEA flat, primrose to pale citrine, white at the margin, reverse same colours. Colonies on OA with a yellow margin, surface black-brown, aerial mycelium sparse, reverse same colours. Colonies on PDA with a distinct and smooth margin, flat, grayish to red, ivory at the edge, reverse darkred.
Additional specimens examined: China: Sichuan Province, Dagu Glacier, N32°8'19" E102°56'13", 2380 m, from soil, 1 May 2017, M-M Wang, culture DG978 = CGMCC3.20233; DG1074 = CGMCC3.20196; N32°15'38" E102°48'15", 3510 m, from soil, 1 May 2017, M-M Wang, culture DG1017 = CGMCC3.20195; N32°14'23" E102°47'7", 3610 m, from water, 1 May 2017, M-M Wang, culture DG1054 = CGMCC3.20234.
Notes — Cadophora indistincta has some similarities with C. ferruginea: red colour colonies on PDA and indistinct collarettes, but the two species are distinguished by conidia shapes: C. indistincta produces cylindrical to oblong conidia (L/W ratio = 2.52) while C. ferruginea produces ellipsoidal conidia (L/W ratio = 1.8).
Cadophora inflata Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837892.
Figure 7
Etymology
Referring to the characteristics of the inflated hyphae.
Type: China: Yunnan Province, Mingyong Glacier, N28°27'24" E98°45'51", 2976 m, from soil, 9 May 2017, M-M Wang, holotype HBU20009, culture ex-type MY759 = CGMCC3.20186.
Mycelium olivaceous or hyaline, septate, branched, smooth-walled, 2–4 µm wide. Hyphal cells often strongly inflated, up to 6–10 µm wide, form chains or microsclerotia-like bodies. Conidiophores always very short or invisible. Conidiogenous cells holoblastic. Conidia hyaline, attached to mycelium, located laterally or terminally, smooth-walled, globular or spathulate, solitary, 2.93–7.05 × 3.00–4.44 µm (mean = 3.91 ± 0.78 × 3.71 ± 0.42 µm, n = 30), L/W ratio = 1.05.
Culture characteristics — Colonies on MEA reaching 28 mm diam, after 14 d at 25°C in the dark, on OA and PDA reaching 47 mm and 37 mm diam, respectively. Colonies on MEA, with an entire margin, flat, milk-white, lacking aerial mycelium, reverse same colours. Colonies on OA with a smooth margin, flat, black in the center, olivaceous to white from middle to edge, reverse same colours. Colonies on PDA with a smooth margin, felty, grey, pale yellow at the margin, reverse grey-brown with a pale buff to white margin.
Notes — Cadophora inflata is characterized by producing chains or microsclerotia-like inflated cells that are similar to C. gamsii and C. echinata which were first described by Maciá-Vicente et al. (2020) and the authors supposed these structures as holoblastic conidia or may just be interpreted as inflated hyphal segments with dormancy functions. The newly described species failed to produce conidia at first, after being induced by slide culture technique, the isolate produced globose or ellipsoidal conidia attaching directly to the hyphae with very short conidiophores that resembled Cadophora orchidicola and thus we suppose that the inflated hyphal cells were just chlamydospores.
Cadophora magna Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837893.
Figure 8
Etymology
Referring to the comparatively huge conidia.
Type: China: Yunnan Province, Mingyong Glacier, N28°27'24" E98°45'51", 2976 m, from soil, 9 May 2017, M-M Wang, holotype HBU20011, culture ex-type MY902 = CGMCC3.20188.
Mycelium hyaline to dark brown, septate, smooth-walled, 1–3 µm, hyphal cells often strongly inflated, variable in shape. Conidiophores brown, smooth-walled, often reduced to conidiogenous cells. Conidiogenous cells phialidic, mostly single, arranged terminally or laterally on the hyphae, cylindrical to navicular, apex wedge, base truncate, smooth-walled, straight or slightly curved, 12.69–20.28 × 2.81–3.84 µm, collarettes funnel-shaped, 1.93–2.98 µm long, opening 2.79–2.88 µm wide. Conidia hyaline, aseptate, smooth-walled, ovoidal or dacryoid to ellipsoidal, upper wedge-shaped, base round, single, straight, 5.24–9.36 × 3.02–4.71 µm (mean = 7.34 ± 0.93 × 3.73 ± 0. 39 µm, n = 30), L/W ratio = 1.97.
Culture characteristics — Colonies on MEA reaching 30 mm diam after 14 d at 25°C in the dark, on OA and PDA reaching 41 mm and 29 mm diam, respectively. Colonies on MEA white, margin covered with ivory and velvety aerial mycelium, reverse white. Colonies on OA with a smooth margin, flat, whitish, pale olive in the centre, reverse same colours. Colonies on PDA creamy to white, reverse same colours.
Notes — The newly described species was isolated from soil samples of Mingyong Glacier and characterized by the huge conidia and strongly inflated hyphal cells.
Cadophora malorum (Kidd & Beaumont) W. Gams
Figure 9
Mycelium brown-black, septate, smooth-walled, branched, 2–3 µm. Conidiophores brown-black, septate, smooth. Conidiogenous cells phialidic, often forming clusters, terminally or laterally on the hyphae, smooth-walled, straight, ampulliform, often 9.47–15.97 × 2.87–3.47 µm, collarettes distinct, collarettes short tubular to funnel-shaped, 1.09–1.98 µm long, opening 1.62–1.94 µm wide. Conidia fuscous, aseptate, smooth-walled, ellipsoidal to elongate-ellipsoidal or subglobose, single, straight, 2.74–4.72 × 1.86–3.40 µm (mean = 3.70 ± 0.51 × 2.46 ± 0.35 µm, n = 30), L/W ratio = 1.50.
Culture characteristics — Colonies on MEA reaching 41 mm diam, after 14 d at 25°C in the dark, on OA and PDA reaching 60 mm and 48 mm diam, respectively. Colonies on MEA with a weakly undulate margin, brown-grey to yellow-brown, reverse same colours. Colonies on OA with a distinct and white margin, olivaceous to dull green, reverse same colours. Colonies on PDA with a distinct margin, felty, gray-brown to gray, reverse isabelline.
Specimen examined: China: Yunnan Province, Yulong Snow Mountain, N27°11'17" E100°22'43", 3362 m, from soil, 7 May 2017, M-M Wang, culture YL412 = CGMCC3.20184.
Notes — Cadophora malorum was the commonest Cadophora species and often isolated as saprobes or pathogens worldwide. Strain YL412 was isolated from soil samples collected from Yulong Snow Mountain and the morphological characteristics were similar with the type.
Cadophora novi-eboraci Travadon, D.P.Lawr., Roon.-Lath., Gubler, W.F. Wilcox, Rolsh. & K. Baumgartner
Figure 10
Mycelium hyaline to brown, septate, smooth-walled, branched, 1–3 µm. Conidiophores hyaline, aseptate, smooth, often solitary. Conidiogenous cells phialidic, terminally or laterally on the hyphae, discrete conidiogenous cells hyaline, smooth-walled, curved or straight, cylindrical to navicular, 6.22–19.90 × 2.38–3.04 µm, collarettes short, tubular, 0.97–1.93 µm long, opening 1.42–1.83 µm wide. Conidia hyaline, aseptate, smooth-walled, elongate-ellipsoidal to cylindrical, straight, 3.90–8.29 × 1.75–2.69 µm (mean = 5.81 ± 1.04 × 2.27 ± 0.26 µm, n = 30), L/W ratio = 2.56.
Culture characteristics — Colonies on MEA reaching 29 mm diam, after 14 d at 25°C in the dark, on OA and PDA reaching 26 mm and 28 mm diam, respectively. Colonies on MEA with an undulate margin, surface beige to ivory, reverse same colours. Colonies on OA with a distinct margin, flat, citrine to pure yellow, white at edge, reverse luteus. Colonies on PDA with a distinct margin, raised, beige to whitish, sometimes covered by floccose aerial mycelium, reverse same colours.
Specimens examined: China: Sichuan Province, Yanzigou Glacier, N29°41'58" E102°0'7", 2620 m, from soil, 29 Apr. 2017, M-M Wang, culture YZ1026 = CGMCC3.20434; YZ1034 = CGMCC3.20190.
Notes — Cadophora novi-eboraci was originally described from decaying wood of Grapevine in North America mainly based on phylogenetic analyses of three nuclear loci (ITS, BT and TEF1-α) (Travadon et al. 2015). Then it was also isolated from Prunus wood or freshwater (Bien & Dam 2020, Lim et al. 2021) and our strains were isolated from soil samples of Yanzigou Glacier in China.
Cadophora psychrophila Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837890.
Figure 11
Etymology
Referring to cold loving.
Type: China: Sichuan Province, Dagu Glacier, N32°14'21" E102°47'5", 3630 m, from soil, 1 May 2017, M-M Wang, holotype HBU20040, culture ex-type DG21 = CGMCC3.20846.
Mycelium black brown or hyaline, septate, smooth-walled, branched, 1–3 µm. Mycelial cell occasionally inflated in the middle, up to 5–8 µm wide, constricted at the septate. Conidiophores black brown or hyaline, septate, mesotonously branched or unbranched. Conidiogenous cells phialidic, hyaline, smooth-walled, tapering toward the tip and slightly constricted at the base, 13.43–23.50 × 2.18–3.84 µm, collarettes distinct and funnel-shaped, 2.82–4.78 µm long, opening 2.61–3.80 µm wide. Conidia hyaline, aseptate, smooth-walled, with subulate tip and round base, single, straight, 4.47–7.77 × 2.09–3.18 µm (mean = 5.53 ± 0.69 × 2.67 ± 0.34 µm, n = 30), L/W ratio = 2.07.
Culture characteristics — Colonies on MEA reaching 19 mm diam, after 14 d at 15°C and 13mm at 25°C in the dark, on OA reaching 28 mm at 15°C and 19 mm at 25°C, and on PDA reaching 25 mm at 15°C and attaining 17 mm at 25°C, respectively. Colonies on MEA raised, glabrous, citrine to primrose, reverse same colours. Colonies on OA with a smooth margin, flat, olive brown in the centre, whitish white at the margin, reverse same colours. Colonies on PDA with a whitish margin, slight raised, pure yellow, reverse same colours.
Additional specimens examined: China: Sichuan Province, Dagu Glacier, N32°14'21" E102°47'5", 3630 m, from soil, 1 May 2017, M-M Wang, culture DG5 = CGMCC3.20845.
Notes — Cadophora psychrophila was characterized by sparse aerial mycelium and slowly growing colony and both isolates in this study showed psychrophilic characteristics that their optimum growth temperature was 15°C.
Cadophora qinghai-tibetana Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837896.
Figure 12
Etymology
Referring to geographical location from which the isolates collected.
Type: China: Sichuan Province, Dagu Glacier, N32°8'19" E102°56'13", 2380 m, from soil, 1 May 2017, M-M Wang, holotype HBU20019, culture ex-type DG1156 = CGMCC3.20193.
Mycelium hyaline or brown-black, septate, smooth-walled, branched, 2–4 µm, often forming coils up to 34.88 µm diam. Conidiophores hyaline, smooth, frequently reduced to conidiogenous cells. Conidiogenous cell phialidic, laterally on the hyphae or hyphae coils, single or in groups of two or three, the mesotonously branched ones often reduced to mere openings with collarettes formed directly on conidiophores, cylindrical or navicular, inflated in the middle and attenuated at the base, hyaline or fuscous, smooth-walled, straight or curved, 6.82–19.94 × 1.97–3.85 µm, collarettes funnel-shaped or absence, 1.61–2.46 µm long, opening 1.57–2.72 µm wide. Sporulation abundant, conidia hyaline, aseptate, smooth-walled, cylindrical to elongate-ellipsoidal, 5.03–7.34 × 1.74–2.72 µm (mean = 6.00 ± 0.66 × 2.13 ± 0.21 µm, n = 30), L/W ratio = 2.82.
Culture characteristics — Colonies on MEA reaching 36 mm diam, after 14 d at 15°C and 19 mm at 25°C in the dark, on OA reaching 40 mm at 15°C and 31 mm at 25°C, and on PDA reaching 35 mm at 15°C and attaining 18 mm at 25°C, respectively. Colonies on MEA with a distinct margin, flat, colony surface creamy to beige, reverse same colours. Colonies on OA with a smooth margin, flat, surface olive black, whitish at the margin, reverse same colours. Colonies on PDA with a distinct and regular margin, aerial mycelium sparse, grey-brown to light brown in the centre, buff to whitish at the margin, reverse same colours.
Additional specimens examined: China: Sichuan Province, Dagu Glacier, N32°13'14" E102°45'29", 4850 m, from soil, 1 May 2017, M-M Wang, culture DG975 = CGMCC3.20232; N32°8'19" E102°56'13", 2380 m, from soil, 1 May 2017, M-M Wang, culture DG1048 = CGMCC3.20191; DG1073 = CGMCC3.20235; DG1087 = CGMCC3.20236; DG1105 = CGMCC3.20197; Sichuan Province, Hailuogou Glacier, N29°34'8" E101°59'36", 3180 m, from soil, 28 Apr. 2017, M-M Wang, culture HL876 = CGMCC3.20437; Yunnan Province, Baima Snow Mountain, N29°23'1" E99°0'20", 4366.2 m, from soil, 10 May 2017, M-M Wang, culture BM327 = CGMCC3.20181; BM360 = CGMCC3.20183; BM523 = CGMCC3.20230; BM816 = CGMCC3.20436; N28°22'59" E99°0'31", 4343 m, from soil, 10 May 2017, M-M Wang, culture BM857 = CGMCC3.20433; Yunnan Province, Mingyong Glacier, N28°27'27" E98°45'49", 2976 m, from soil, 9 May 2017, M-M Wang, culture MY474 = CGMCC3.20185; N28°27'28" E98°45'43", 3067 m, from soil, 9 May 2017, M-M Wang, culture MY492 = CGMCC3.20847; MY527 = CGMCC3.20848; MY588 = CGMCC3.20849; MY589 = CGMCC3.20850; MY873 = CGMCC3.20231; Yunnan Province, Yulong Snow Mountain, N27°10'55" E100°19'87", 4531 m, from soil, 7 May 2017, M-M Wang, culture YL73 = CGMCC3.20228; N27°11'17" E100°22'43", 3362 m, from water, 7 May 2017, M-M Wang, culture YL305 = CGMCC3.20435; YL319 = CGMCC3.20229; YL357 = CGMCC3.20182; YL414 = CGMCC3.20194.
Notes — More than half of the isolates in this study were identified as Cadophora qinghai-tibetana and they were isolated from soil and water samples of Yulong Glacier, Mingyong Glacier, Baima Snow Mountain in Yunnan Province and Dagu Glacier in Sichuan Province. Strains of YL73 (from Yulong Snow Mountain), DG1048, DG1073, DG1087, DG1105 and DG1156 (from Dagu Glacier), MY527, MY588, MY589 and MY873 (from Mingyong Glacier) showed psychrophilic characteristics that they had optimum growth temperature at 15°C while the others had optimum growth temperature at 25°C. C. qinghai-tibetana had typical phialidic conidiogenesis and produced cylindrical to elongate-ellipsoidal conidia that were common in many Cadophora species, but all strains of this species formed a well-supported clade which was distinct from others in the multigene phylogenetic tree (Fig. 4).
Cadophora yulongensis Q-M Wang, B-Q Zhang & M-M Wang, sp. nov.
MycoBank No.: MB837894.
Figure 13
Etymology
Referring to Yulong Snow Mountain, the geographic origin of this species.
Type: China: Yunnan Provinces, Yulong Snow Mountain, N27°10'52" E100°19'84", 4531 m, from soil, 7 May 2017, M-M Wang, holotype HBU20010, culture ex-type YL814 = CGMCC3.20187.
Mycelium hyaline, septate, smooth-walled, branched, 1–3 µm wide. Conidiophores hyaline, smooth, often reduced to conidiogenous cells. Conidiogenous cells phialidic, located laterally or terminally, cylindrical or navicular, apex wedge, base truncate, hyaline, smooth-walled, straight or bent, 11.43–25.52 × 1.61–3.10 µm, collarettes evident, 2.10–4.54 µm long, opening 1.59–2.45 µm wide. Conidia hyaline, aseptate, smooth-walled, cylindrical, sporulation abundant, single, straight, 4.48–6.91 × 1.36–2.51 µm (mean = 5.50 ± 0.63 × 1.89 ± 0.29 µm, n = 30), L/W ratio = 2.91.
Culture characteristics — Colonies on MEA reaching 36 mm diam, after 14 d at 25°C in the dark, on OA and PDA reaching 38 mm and 28 mm diam, respectively. Colonies on MEA pale pink to whitish, white at the margin, reverse same colours. Colonies on OA black-gray with a grey-white margin, reverse same colours. Colonies on PDA grey to brown, margin wheat, cottony, reverse olive-brown to yellowish from centre to margin.
Notes — At the beginning, Cadophora yulongensis failed to produce conidia on MEA, OA and PDA medium. Other efforts including Pine needle medium culturing and H2O2 treatment (Xu et al, 2009) were also failed to induce sporulation until using slide culture technique. In the multigene phylogenetic tree (Fig. 4), C. yulongensis was closely related to lineages formed by species with holoblastic conidiogenesis, but this species was characterised by long cylindrical phialides and high conidium length/width ratio (2.91).