Research Article |
Corresponding author: Peter E. Mortimer ( peter@mail.kib.ac.cn ) Academic editor: Cecile Gueidan
© 2019 De-Ping Wei, Dhanushka N. Wanasinghe, Kevin D. Hyde, Peter E. Mortimer, Jianchu Xu, Yuan-Pin Xiao, Chitrabhanu S. Bhunjun, Chaiwat To-anun.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Wei D-P, Wanasinghe DN, Hyde KD, Mortimer PE, Xu J, Xiao Y-P, Bhunjun CS, To-anun C (2019) The genus Simplicillium. MycoKeys 60: 69-92. https://doi.org/10.3897/mycokeys.60.38040
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Simplicillium species have a wide host range and an extensive distribution. Some species are associated with rusts, as well as other plant pathogenic fungi and play an important role in biological control. In this study, two specimens of Simplicillium were collected from Chiang Mai Province, Thailand. Simplicillium formicae sp. nov. was isolated from an infected ant and S. lanosoniveum from Ophiocordyceps unilateralis which is a new host record. Species were initially identified using ITS gene sequences and confirmed using morphology coupled with phylogenetic analyses of a combined nrLSU, nrSSU, TEF and RPB1 dataset. Simplicillium formicae differs from other species in the genus by the presence of flask-shaped synnemata and phialides with intercalary nodes. Simplicillium lanosoniveum resembles other collections of the species by its completely solitary, tapering phialides and globose to ellipsoidal conidia which adhere in a slimly head. A key to species of Simplicillium is also provided.
new species, Thailand, ant fungi, taxonomy, phylogeny
Simplicillium species have a wide distribution and are considered as mammal and plant-parasitic, symbiotic, entomopathogenic, fungicolous and nematophagous fungi, as they have a broad spectrum of hosts and substrates, such as insects, plants, rusts, nematodes, human nails, canine tissues and mushrooms, Chroococcus sp., soil, freshwater, marine and terrene environments (
Recent studies have shown that Thailand supports an amazing fungal diversity with numerous new species that have the potential for biotechnological application (
The Mushroom Research Centre (MRC) is a disturbed rainforest located in Chiang Mai Province, Thailand (
For long-term deposit, these two specimens were dried with allochroic silica gel to protect them from contamination of opportunistic fungi and to retain the informative taxonomic characters. The macro-morphological characters were observed with a stereoscope (Olympus SZ61) and the micro-morphological features were examined with a compound microscope (Nikon ECLIPSE Ni). Important characteristics such as mycelium, phialides and conidia were captured with a digital camera (Canon EOS 600D). Measurements of perithecia, synnemata, phialides and conidia were taken using the Tarosoft (R) Image Frame Work programme and the images used were processed with Adobe Photoshop CS3 Extended v. 10.0 (Adobe, San Jose, CA).
DNA was extracted from fresh mycelium of isolates MFLUCC 18–1379 and MFLUCC 18–1385 and from stromal tissue of ant fungus HKAS 102447 (the host of isolate MFLUCC 18–1385) using a DNA extraction kit (Biospin Fungus Genomic DNA Extraction Kit, BioFlux, China), following the instructions of the manufacturer. Extracted DNA was stored at 4 °C for use in regular work and duplicated at –20 °C for long-term storage. The internal transcribed spacer (ITS1-5.8S-ITS2, ITS) was amplified with primer ITS4 and ITS5 (
The raw sequences were verified with Finch TV version 1.4.0 (
Strains and GenBank accession numbers from related references used in multi-gene tree.
Taxon | Voucher no. | Host/substrate | SSU rRNA | LSU rRNA | tef1 | rpb1 | Reference |
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Akanthomyces tuberculata | OSC 111002 | Lepidoptera | DQ522553 | DQ518767 | DQ522338 | DQ522384 |
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Aschersonia badia | BCC 8105 | Scale insect | DQ522537 | DQ518752 | DQ522317 | DQ522363 |
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Aschersonia placenta | BCC 7957 | Scale insect | DQ522538 | DQ518753 | DQ522318 | DQ522364 |
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Balansia henningsiana | GAM 16112=AEG96-27a | Panicum sp. | AY545723 | AY545727 | AY489610 | AY489643 |
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Balansia pilulaeformis | AEG 94-2 | Poaceae | AF543764 | AF543788 | DQ522319 | DQ522365 |
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Claviceps fusiformis | ATCC 26019 | Poaceae | DQ522539 | U17402 | DQ522320 | DQ522366 |
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Claviceps paspali | ATCC 13892 | Poaceae | U32401 | U47826 | DQ522321 | DQ522367 |
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Claviceps purpurea | GAM 12885 | Dactylis glomerata | AF543765 | AF543789 | AF543778 | AY489648 |
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Cordyceps farinosa | OSC 111005 | Lepidoptera pupa | DQ522558 | DQ518773 | DQ522348 | DQ522394 |
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Cordyceps heteropoda | OSC 106404 | Cicada | AY489690 | AY489722 | AY489617 | AY489651 |
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Cordyceps militaris | OSC 93623 | Lepidoptera | AY184977 | AY184966 | DQ522332 | DQ522377 |
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Cordyceps ophioglossoides | OSC 106405 | Elaphomyces sp. | AY489691 | AY489723 | AY489618 | AY489652 |
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Cordyceps pruinosa | ARSEF 5413 | Iragoides fasciata | AY184979 | AY184968 | DQ522351 | DQ522397 |
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Cordyceps scarabaeicola | ARSEF 5689 | Scarabaeidae pupa | AF339574 | AF339524 | DQ522335 | DQ522380 |
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Cordyceps tenuipes | OSC 111007 | Lepidoptera pupa | DQ522559 | DQ518774 | DQ522349 | DQ522395 |
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Drechmeria balanoides | CBS 250.82 | Nematoda | AF339588 | AF339539 | DQ522342 | DQ522388 |
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Drechmeria gunnii | OSC 76404 | Lepidoptera larva | AF339572 | AF339522 | AY489616 | AY489650 |
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Drechmeria sinensis | CBS 567.95 | Nematoda | AF339594 | AF339545 | DQ522343 | DQ522389 |
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Engyodontium aranearum | CBS 309.85 | Spider | AF339576 | AF339526 | DQ522341 | DQ522387 |
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Epichloë typhina | ATCC 56429 | Festuca rubra | U32405 | U17396 | AF543777 | AY489653 |
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Hypocrella nectrioides | GJS 89-104 | Scale insect | U32409 | U47832 | DQ522347 | DQ522393 |
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Hypocrella schizostachyi | BCC 14123 | Scale insect | DQ522557 | DQ518771 | DQ522346 | DQ522392 |
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Lecanicillium antillanum | CBS 350.85 | Agaric | AF339585 | AF339536 | DQ522350 | DQ522396 |
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Lecanicillium lecanii | CBS 101247=IMI 304807 | Coccus viridis | AF339604 | AF339555 | DQ522359 | DQ522407 |
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Lecanicillium wallacei | CBS 101237=IMI 331549 | Lepidoptera | AY184967 | EF469073 | EF469102 |
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Metacordyceps chlamydosporia | CBS 101244 | Mollusca | DQ522544 | DQ518758 | DQ522327 | DQ522372 |
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Metacordyceps taii | ARSEF 5714 | Lepidoptera | AF543763 | AF543787 | AF543775 | DQ522383 |
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Metapochonia goniodes | CBS 891.72 | Nematoda | AF339599 | AF339550 | DQ522354 | DQ522401 |
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Metarhizium album | ARSEF 2082 | Cofana spectra | DQ522560 | DQ518775 | DQ522352 | DQ522398 |
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Metarhizium flavoviride | ARSEF 2037 | Nilaparvata lugens | AF339580 | AF339531 | DQ522353 | DQ522400 |
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Metarhizium majus | ARSEF 3145 | Oryctes rhinoceros | AF339579 | AF339530 | AF543774 | DQ522399 |
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Myriogenospora atramentosa | AEG 96-32 | Andropogon virginicus | AY489701 | AY489733 | AY489628 | AY489665 |
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Ophiocordyceps acicularis | OSC 128580 | Coleoptera | DQ522543 | DQ518757 | DQ522326 | DQ522371 |
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Ophiocordyceps aphodii | ARSEF 5498 | Coleoptera | DQ522541 | DQ518755 | DQ522323 |
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Ophiocordyceps brunneipunctata | OSC 128576 | Coleoptera | DQ522542 | DQ518756 | DQ522324 | DQ522369 |
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Ophiocordyceps irangiensis | OSC 128577 | Ant | DQ522546 | DQ518760 | DQ522329 | DQ522374 |
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Ophiocordyceps irangiensis | OSC 128578 | Ant | DQ522556 | DQ518770 | DQ522345 | DQ522391 |
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Ophiocordyceps melolonthae | OSC 110993 | Scarabaeidae larva | DQ522548 | DQ518762 | DQ522331 | DQ522376 |
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Ophiocordyceps nutans | OSC 110994 | Stink bug | DQ522549 | DQ518763 | DQ522333 | DQ522378 |
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Ophiocordyceps ravenelii | OSC 110995 | Phyllophaga sp. | DQ522550 | DQ518764 | DQ522334 | DQ522379 |
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Ophiocordyceps sphecocephala | OSC 110998 | Wasp | DQ522551 | DQ518765 | DQ522336 | DQ522381 |
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Ophiocordyceps stylophora | OSC 111000 | Elateridae grub | DQ522552 | DQ518766 | DQ522337 | DQ522382 |
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Ophiocordyceps unilateralis | OSC 128574 | Ant | DQ522554 | DQ518768 | DQ522339 | DQ522385 |
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Ophiocordyceps variabilis | ARSEF 5365 | Diptera larva | DQ522555 | DQ518769 | DQ522340 | DQ522386 |
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Rotiferophthora angustispora | CBS 101437 | Rotifera | AF339584 | AF339535 | AF543776 | DQ522402 |
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Simplicillium calcicola | LC5586 = CGMCC3.17943 | Calcaire | KY883301 | KU746752 | KX855252 |
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Simplicillium lamellicola | CBS 116.25 | Agaricus bisporus | AF339601 | AF339552 | DQ522356 | DQ522404 |
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Simplicillium lanosoniveum | CBS 704.86 | Hemileia vastatrix | AF339602 | AF339553 | DQ522358 | DQ522406 |
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Simplicillium obclavatum | CBS 311.74 | Air above sugarcane filed | AF339567 | AF339517 | EF468798 |
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Tolypocladium fractum | OSC 110990 | Elaphomyces sp. | DQ522545 | DQ518759 | DQ522328 | DQ522373 |
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Tolypocladium japonicum | OSC 110991 | Elaphomyces sp. | DQ522547 | DQ518761 | DQ522330 | DQ522375 |
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Torrubiella ratticaudata | ARSEF 1915 | Euophrys sp. | DQ522562 | DQ518777 | DQ522360 | DQ522408 |
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Verticillium epiphytum | CBS 384.81 | Hemileia vastatrix | AF339596 | AF339547 | DQ522361 | DQ522409 |
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The generated sequences of each gene region were aligned separately with representative sequences using MAFFT v. 7 web server (http://mafft.cbrc.jp/alignment/server) (
Phylogenetic trees were visualised with FigTree v1.4.0 (
The combined four gene dataset comprised 60 taxa from three families (Cordycipitaceae, Ophiocordycipitaceae and Clavicipitaceae) (Table
Phylogram generated from maximum likelihood analysis based on combined SSU, LSU, TEF and RPB1 sequence data. Bootstrap values for maximum likelihood (ML, left) and maximum parsimony (MP, right) equal to or greater than 50% and Bayesian posterior probabilities (BYPP, middle) equal to or greater than 0.90 are placed nearby the note. The newly generated sequences are indicated in red bold.
The ITS dataset comprised 49 taxa from all Simplicillium species that are currently available in GenBank (Figure
Phylogram generated from maximum likelihood analysis based on ITS sequence data. Bootstrap values for maximum likelihood (ML, left) and maximum parsimony (MP, right) equal to or greater than 50% and Bayesian posterior probabilities (BYPP, middle) equal to or greater than 0.90 are placed nearby the branches, respectively. The newly generated sequences are indicated in red bold and the type species are highlighted in black bold.
The multi-gene phylogenetic analyses showed that our isolates MFLUCC 18-1379 and MFLUCC 18–1385 grouped with the remaining Simplicillium species with strong support (100% ML, 1.00 BYPP, 100% MP, Figure
Hyperparasitic on rusts or parasitic on nematodes or occurring in soil. Asexual morph: Mycelium thin, hyaline, septate, branched, smooth-walled. Phialides arising from prostrate aerial hyphae or rope-like and flask-shaped synnemata, typically solitary, rarely in whorls of 2–3, gradually tapering towards the apex, elongate, slender, smooth-walled, phialidic. Conidia hyaline, oval, spindle-shaped, cylindrical, subglobose to ellipsoidal, fusoid to filiform, straight to curved, smooth-walled. Conidia commonly form in small globose heads, sometimes in branched, unbranched, zigzag or imbricate chains, occasionally in sympodial proliferation with cylindrical conidium-bearing denticles. Colonies of species in this genus are usually fast growing, reaching 10–38 mm within 10 days on PDA, white, reverse brownish-cream to pale yellow, margin entire, cottony, fluffy or floccose. Some species produce yellow or orange pigment. Crystals are commonly present in the agar. Sexual morph: Torrubiella (
In this study, we introduce a new species and a new host species as described below.
the epithet refers to its host–ant.
HKAS 102459; living culture: MFLUCC 18–1379.
Parasitic on ant (Formicidae). Asexual morph: Hyphomycetous. Mycelium rarely septate, hyaline, smooth-walled, covering the whole body of the ant host. Synnemata 250–350 × 65–100 (xˉ = 300 × 86, n = 10) µm, forming at the head region of ant host in circular arrangement, flask-shaped, hyaline to yellowish, composed of dense hyphae, somehow curved. Phialides 25–100 × 0.5–1.5 (xˉ = 49 × 1.1, n = 20) µm, arising from procumbent hyphae or synnemata, blastic, enteroblastic, phialidic, monophialidic, discrete, terminal, unbranched, solitary, aseptate, hyaline, smooth-walled, slender, occasionally a swollen node present. Conidia 2–3.5 × 1.5–2.5 (xˉ = 2.6 × 2, n = 30) µm, globose to ellipsoidal, hyaline, one-celled, smooth-walled, round at both ends, adhering in slimy head on the tip of phialides. Sexual morph: Undetermined.
The colonies were rapid-growing on PDA medium, reaching a diameter of 2.5–3 (xˉ = 2.6, n = 9) cm, in 13 days at 22 °C, entire margin, circular, velvety and white from above, with radial crack and primrose-yellow on reverse. In vitro, Synnemata absent. Phialides 25–75 × 0.4–0.6 (xˉ = 50 × 0.55, n = 10) µm, arising from procumbent hyphae, blastic, enteroblastic, phialidic, discrete, terminal, unbranched, solitary, aseptate, hyaline, smooth-walled, relatively slender and long. Conidia 1.5–3 × 1.5–2.5 (xˉ = 2.3 × 1.7, n = 100) µm, hyaline, globose to ellipsoidal, aseptate, smooth-walled, slightly guttulate, adhering in slimy head on the tip of phialides.
THAILAND, Chiang Mai Province, Mushroom Research Centre, on an adult ant, 1 April 2018, Deping Wei, MRC18040102 (holotype: HKAS 102459; ex-type living culture: MFLUCC 18–1379). Sequences generated from this strain have been deposited in GenBank with accession numbers: SSU = MK765046, LSU = MK766512, ITS = MK766511, TEF = MK926451, RPB1 = MK882623.
Isolate MFLUCC 18–1379 has a close phylogenetic relationship with Simplicillium obclavatum, based on ITS sequence analysis. The new isolate is similar to Simplicillium obclavatum in terms of shape and dimensions of the conidia with slender phialides tapering towards the apex. However, they have a different conidial arrangement, by Simplicillium obclavatum having short-imbricate chains, whereas the new fungus has subglobose to globose head. There are numerous synnemata in a circular arrangement which can be observed in our isolate and those are absent in Simplicillium obclavatum. The comparisons of ITS sequences between our isolate MFLUCC 18–1379 and ex-type strain of Simplicillium obclavatum (CBS 311.74) show 23 bp differences within 550 bp (4.2%). Thereby, we identify our isolates as a new species according to
Cephalosporium lanosoniveum J.F.H. Beyma, Antonie van Leeuwenhoek 8: 121 (1942) (Basionym)
Netherlands, on hair of Cibotium schiedei in greenhouse, 1942, F.H. van Beyma, CBS123.42.
Saprophytic on Ophiocordyceps unilateralis. Asexual morph: Hyphomycetous. Mycelium aseptate, hyaline, smooth-walled. Phialides 20–40 × 1.1–2 (xˉ = 30 × 1.6, n = 20) µm, arising from the prostrate mycelium, blastic, enteroblastic, phialidic, monophialidic, discrete, terminal, aseptate, hyaline, smooth-walled, solitary, tapering toward the apex. Conidia 2–4.5 × 1–3 (xˉ = 3 × 1.8, n = 60) µm, hyaline, amerospores, globose to ellipsoidal, smooth-walled, adhering in globose to ellipsoidal head at the apex of phialides. Sexual morph: Undetermined.
The colonies on PDA medium were rapid-growing, reaching a diam. of 5.5 cm in 30 days at 22 °C, white, entire margin, velvety, with radial cracks and primrose-yellow on the reverse.
Host and distribution: Saprophytic on fungi, endophytic or symbiotic or pathogenic on plant, parasitic on rust, nematode and insect, occurring on soil, animal hair or human bronchoalveolar lavage fluid, with a cosmopolitan distribution (see Table
Distribution, host and available sequence data of Simplicillium lanosoniveum strains.
Species | Strain no. | Host and habitat | Origin | Available gene region | Morphological description | Reference |
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S. lanosoniveum | CBS123.42 | Hair of Cibotium schiedei (Plant) | Netherland | ITS, LSU | GenBank; |
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Cs0701 | Salvinia molesta (Plant) | Taiwan | ITS | √ |
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PSU-ES104 | Enhalus acoroides (Plant) | Trang Province, Thailand | ITS |
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CBS 531.72 | Salvinia rotundifolia (Plant) | USA | ITS |
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Tr3 | Salvia miltiorrhiza (Plant) | China | ITS | GenBank | ||
YLAC-5 | Endophytic on Inula aconitum (Plant) | China | ITS | GenBank | ||
Endophytic on seaweed (Plant) | India | SSU | GenBank | |||
E1, E3, E5 | Endophytes of Sophora alopecuroides (Plant) | Ningxia, China | SSU, ITS | √ |
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GA-B1 | Grewia asiatica (Plant) | Shivalik region, Jammu, India | SSU | GenBank | ||
IMI 303103b | Hemileia vastatrix (Rust) | Colombia | ITS, SSU |
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AMH 9654 | Rust pustules on leaves of Elaeagnus sp. | India | LSU, ITS | √ |
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D082307-2A | Soybean rust | Louisiana | ITS | √ |
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vecl-02 | Rust of Eleagnus latifolia | India | ITS | GenBank | ||
vecl-01 | Rust of Eleagnus latifolia | India | ITS | GenBank | ||
CBS 704.86 | Hemileia vastatrix (Rust) | Venezuela | ITS, SSU, LSU, TEF, RPB1, RPB2, ATP | √ | GenBank; |
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S-599 | Coleosporium plumeriae (Rust) | Campos dos Goytacazes, GJ, Brazil | ITS |
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D082307-2A-GFP15 | Phakopsora pachyrhizi (Rust) | Florida, USA | √ |
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HKAS 102447 | Ophiocordyceps unilateralis (Fungi) | Chiang Mai, Thailand | SSU, LSU, ITS, TEF, RPB1 | √ | This study | |
TYL001 | Pseudaulacaspis pentagona (Insect) | Shanxi Province, China | ITS, SSU | √ |
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SSBG2 | Coccus hesperidum (Insect) | The South-Siberian Botanical Garden, Russia | ITS | √ |
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TAMA 173 | Aphidoidea sp. (Insect) | Ibaraki, Japan | ITS |
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CHE-CNRCB 373 | Diaphorina citri (Insect) | Colima, USA | ITS |
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ARSEF 8822 | Culicid (Insect) | Tanzania |
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ARSEF7550 | Coccoidea (Insect) | Brazil | TEF | GenBank | ||
1T9BA | Tick (Insect) | New York, USA | ITS |
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Btab03 | Bemisia tabaci (Insect) | South Korea | ITS | GenBank | ||
113-8 | Mosquitoes (Insect) | Japan | ITS |
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S. lanosoniveum | 7S | Heterodera schachtii (Nematode) | Iran | ITS | GenBank | |
Hair of giant panda (Animal) | China | ITS | GenBank | |||
2502 | Bronchoalveolar lavage fluid (Human) | China | ITS | GenBank | ||
41559-3 | Cave and mine | New York State, USA | ITS, LSU | GenBank | ||
CBS 321.72 | Malaysia | SSU, LSU, ITS | Genbank; |
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CBS 322.72 | Malaysia | ITS | GenBank |
THAILAND, Chiang Mai Province, Mushroom Research Centre, on Ophiocordyceps unilateralis, 19 February 2018, Deping Wei, MRC18021901 (HKAS 102447; living culture: MFLUCC 18–1385). Sequences generated from this strain have been deposited in GenBank with accession numbers: SSU = MK752791, LSU = MK752849, ITS = MK752683, TEF = MK926450, RPB1 = MK882622.
Our isolate MFLUCC 18–1385 colonised on a decayed Ophiocordyceps unilateralis with white hyphae. In a thorough examination of the Ophiocordyceps unilateralis host, we found the phialides and conidia of our isolate grown on the surface of the host (Figure
Simplicillium formicae (from HKAS 102459, holotype) a superficial hyphae associated with the ant host b–e flask-shaped synnemata f–k phialides bearing conidia l–p conidia. Scale bars: 1000 µm (a); 500 µm (b); 100 µm (d); 30 µm (e, f); 15 µm (j, k); 10 µm (l–p) (e stained with cotton blue solution).
Simplicillium formicae (MFLUCC 18–1379, ex-type living culture) a upper and reverse view of cultures on PDA after 30 days e–g phialides indicated with black arrow c, d, h–j conidial mass on the tip of phialides k–m conidia. Scale bars: 10 µm (c, d, f, g); 20 µm (e); 3 µm (h–j); 1 µm (k–m) (e–j stained with cotton blue solution).
Simplicillium lanosoniveum (a–f from HKAS 102447, g–r from MFLUCC 18–1385) a host (Ophiocordyceps unilateralis); b, c hyphae associated with host indicated with black arrows g, h upper and reverse view of cultures on PDA after 40 days incubation i–l conidial mass on the tip of phialides m–o phialides bearing conidia p–r conidia. Scale bars: 15 µm (i–m); 10 µm (d–f, n, o); 3 µm (p–r) (i, l–n stained with cotton blue solution).
Parasitic on ants (Formicidae). Sexual morph: Stromata up to 14 mm in length, 0.5 mm wide in the broadest part, cylindrical, brown, slightly tapering towards the apex, single, piercing through the dorsal neck region of the ant host. Ascomatal cushion hemisphere, up to 1.2 mm in diam., laterally attaching to the erect stroma stalk, dark brown, with ostioles protruding from the cushions. Perithecia 200–400 × 50–120 (xˉ = 294 × 81, n = 10) µm, sub-immersed, flask-shaped. Asci and ascospores were too old to observe their features. Asexual morph: Undetermined.
This collection was already decayed and was colonised by other fungi which we introduced as a new host record of Simplicillium lanosoniveum from Thailand. The outline of this specimen was intact, while its asci and ascospores were too old to analyse. We retrieved DNA through direct sequencing from the stromal tissue.
Sequences generated from this specimen have been deposited in GenBank with accession numbers: SSU = MK752759, LSU = MK752812, ITS = MK752874. The herbarium material is deposited at KUN herbarium, Yunnan Province, China. In the multi-gene phylogenetic tree, this collection groups with Ophiocordyceps unilateralis (OSC 128574) with a strongly supported bootstrap value (100% ML, 1.00 BYPP, 100% MP, Figure
1a | Conidia formed in sympodia | S. sympodiophorum |
1b | Conidia solitary, borne on the tip of phialides | S. calcicole |
1c | Conidia aggregate in chains | 2 |
1d | Conidia aggregate in subglobose to ellipsoidal heads | 3 |
1e | Conidia aggregate in globose heads | 4 |
2a | Conidia 2.5–3.5 × 1–2 µm, obclavate to ellipsoidal, formed in short imbricate chains | S. obclavatum |
2b | Conidia 3.5–5.0 × 1.0–1.5 μm, oval, ellipsoidal or cylindrical, formed in vertical chains | S. chinense |
2c | Conidia 7.2–12.5 × 1 µm, long, fusiform to short filiform, hyaline, straight to curved, formed in vertical chains | S. filiforme |
3a | Phialides 15–50 × 0.7–1.0 µm, colonies light to dark-brown reverse on PDA, usually with yellow pigment diffusing into the agar | S. lamellicola |
3b | Phialides 11–44 (–70) × 1.0–2.4 µm, colonies cream-coloured reverse on PDA, no diffused pigment | S. coffeanum |
4a | Present flask-shaped synnemata | S. formicae |
4b | Synnemata absent | 5 |
5a | Conidia cylindrical | 6 |
5b | Conidia globose to subglobose or ellipsoidal | 7 |
6a | Phialides 23–53 × 1.2–2.0 µm, long | S. cylindrosporum |
6b | Phialides 17–32 × 1.2–2.0(–2.5) µm, short | S. aogashima |
7a | Phialides 35–75 × 1.2–3.0 µm, conidia 4.5–6.0 × 2.5–3.5 µm, colonies light yellow to deep tawny in reverse view on PDA | S. lanosoniveum var. tianjinensis |
7b | Phialides 15–39 × 0.7–1.9 µm, conidia 1.5–3 × 0.7–1.3 µm, colonies brownish-cream to pale yellow reverse on PDA | S. lanosoniveum |
7c | Phialides 11–31(–47) × 1.0–1.7 µm, conidia 2.0–3.5 × 1.8–2.5(–2.8) µm, colonies brown reverse on PDA | S. minatense |
7d | Phialides (15–)20–42(–50) × 1.0–2.3 µm; conidia 2.3–4.0(–4.5) × 1.5–3.3 µm, colonies brownish-orange to brown reverse on PDA | S. subtropicum |
A new species Simplicillium formicae and a new host record species Simplicillium lanosoniveum from Ophiocordyceps unilateralis were introduced, based on phylogenetic analyses and morphological evidence. The host and distribution of S. lanosoniveum was summarised and a key to Simplicillium was provided.
We are grateful to the Thailand Research Fund (TRF) grant no DBG6080013 entitled ‘the future of specialist fungi in a changing climate: baseline data for generalist and specialist fungi associated with ants, Rhododendron species and Dracaena species’’ for its financial support. Dhanushka Wanasinghe would like to thank CAS President’s International Fellowship Initiative (PIFI) for funding his postdoctoral research (number 2019PC0008) and the 64th batch of China Postdoctoral Science Foundation (grant no.: Y913083271). Peter E. Mortimer and D.N. Wanasinghe thank the National Science Foundation of China and the Chinese Academy of Sciences for financial support under the following grants: 41761144055, 41771063 and Y4ZK111B01. We acknowledge the Kunming Institute of Botany for providing the laboratories and instruments for molecular work. We appreciate the Centre of Excellence in Fungal Research (Mae Fah Luang University) for providing the grant support for collecting trips. We thank Milan Samarakoon and Dr. Sajeewa Maharachchikumbura for their assistance with the phylogenetic analyses and Dr. Shaun Pennycook for his help with the nomenclature of the novel species.