Research Article |
Corresponding author: Yan-Feng Han ( swallow1128@126.com ) Academic editor: Andrew Miller
© 2022 Xin Li, Zhi-Yuan Zhang, Yu-Lian Ren, Wan-Hao Chen, Jian-Dong Liang, Ji-Mei Pan, Jian-Zhong Huang, Zong-Qi Liang, Yan-Feng Han.
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:
Li X, Zhang Z-Y, Ren Y-L, Chen W-H, Liang J-D, Pan J-M, Huang J-Z, Liang Z-Q, Han Y-F (2022) Morphological characteristics and phylogenetic evidence reveal two new species of Acremonium (Hypocreales, Sordariomycetes). MycoKeys 91: 85-96. https://doi.org/10.3897/mycokeys.91.86257
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Using chicken feathers as bait, Acremonium globosisporum sp. nov. and Acremonium curvum sp. nov. were collected from the soil of Yuncheng East Garden Wildlife Zoo and Zhengzhou Zoo in China. They were identified by combining the morphological characteristics and the two-locus DNA sequence (LSU and ITS) analyses. In the phylogenetic tree, both new species clustered into separate subclades, respectively. They were different from their allied species in their morphology. The description, illustrations, and phylogenetic tree of the two new species were provided.
Acremonium, filamentous fungi, phylogeny, taxonomy
The genus Acremonium Link, established in 1929, with A. alternatum Link as the type species, is one of the largest and most complex genera of asexually typified. The morphological characteristics consist of hyphae septate, mostly tapered and lateral phialides, produced singly or in small groups, and unicellular conidia produced in mucoid heads or unconnected chains (
Due to the poor differentiation of asexual forms of the genus Acremonium, it is difficult to identify species only by morphological differences. To address this issue, there are many unidentified and suspicious species that require further phylogenetic analysis. To date, many isolates of Acremonium spp. lack the gene loci such as SSU, TEF 1-α and RPB2 (Table
In the present study, two new species of Acremonium were identified in a survey of keratinolytic fungi from China, which were enriched by the baiting technique. We provided a description, illustrations, and phylogenetic tree for the two new species.
Soil samples were collected from Yuncheng East Garden Wildlife Zoo (35°6'26"N, 111°4'24"E) (three isolates), Yuncheng City, Shanxi Province and Zhengzhou Zoo (34°47'20"N, 113°40'41"E) (one isolate), Zhengzhou City, Henan Province, China by Yu-Lian Ren on July 2021. We collected 3–10 cm below the soil surface, placed the samples in sterile Ziploc plastic bags (Kaixin Biotechnology, Guizhou, China), and transported them to the laboratory (
Then, the 2 g samples were weighed in a conical flask with glass beads containing 20 mL sterile water and mixed evenly by eddy shock for 10 min. Next, 1 mL samples were mixed evenly in 9 mL sterile water in a sterile environment and diluted to 10-3. Then, 1 mL 10-3 samples were put into a sterile petri dish, and SDA medium containing 50 mg/L penicillin and 50 mg/L streptomycin was added and mixed. The target strains were isolated. The purified strains were transferred to PDA, OA, and MEA plates for dark culture at 25 °C for 7 days. Microscopic features were examined by making direct wet mounts with 25% lactic acid on PDA, with a light microscope.
The cultures were placed to slowly dry at 50 °C to produce the dried holotype. The dried holotype was deposited in the Mycological Herbarium of the Institute of Microbiology, Chinese Academy of Sciences, Beijing, China (
We used a 5% chelex-100 solution for total genomic DNA extraction. ITS1/ITS4 (
Species | Strains | LSU | ITS | SSU | TEF 1-α | RPB2 |
---|---|---|---|---|---|---|
Acremonium alcalophilum | CBS 114.92T | JX158443 | DQ825967 | JX158486 | JX158399 | JX158465 |
Acremonium alternatum | CBS 407.66T | HQ231988 | HE798150 | |||
Acremonium alternatum | CBS 831.97 | HQ231989 | ||||
Acremonium arthrinii | MFLU 18-1225T | MN036334 | MN036335 | MN038169 | ||
Acremonium behniae | CBS 146824T | MW175400 | MW175360 | |||
Acremonium biseptum | CBS 750.69T | HQ231998 | ||||
Acremonium blochii | CBS 993.69 | HQ232002 | HE608636 | |||
Acremonium borodinense | CBS 101148T | HQ232003 | HE608635 | |||
Acremonium brachypenium | CBS 866.73T | HQ232004 | AB540570 | |||
Acremonium camptosporum | CBS 756.69T | HQ232008 | HQ232186 | |||
Acremonium cavaraeanum | CBS 101149T | HF680202 | HF680220 | |||
Acremonium cavaraeanum | CBS 111656 | HF680203 | HF680221 | |||
Acremonium cavaraeanum | CBS 758.69 | HQ232012 | HF680222 | |||
Acremonium cerealis | CBS 207.65 | HQ232013 | ||||
Acremonium cerealis | CBS 215.69 | HQ232014 | ||||
Acremonium chiangraiense | MFLUCC 14-0397T | MN648329 | MN648324 | |||
Acremonium chrysogenum | CBS 144.62T | HQ232017 | HQ232187 | |||
Acremonium chrysogenum | CBS 401.65 | MH870276 | MH858636 | |||
Acremonium citrinum | CBS 384.96T | HF680217 | HF680236 | |||
Acremonium dimorphosporum | CBS 139050T | LN810506 | LN810515 | |||
Acremonium exiguum | CBS 587.73T | HQ232035 | ||||
Acremonium exuviarum | UAMH 9995T | HQ232036 | AY882946 | |||
Acremonium felinum | CBS 147.81T | AB540488 | AB540562 | |||
Acremonium flavum | CBS 596.70T | HQ232037 | HQ232191 | |||
Acremonium flavum | CBS 316.72 | MH872204 | MH860487 | |||
Acremonium fuci | CBS 112868T | AY632653 | ||||
Acremonium fuci | CBS 113889 | AY632652 | ||||
Acremonium fusidioides | CBS 109069 | HF680204 | HF680223 | |||
Acremonium fusidioides | CBS 991.69 | HF680211 | HF680230 | |||
Acremonium fusidioides | CBS 840.68T | HQ232039 | FN706542 | |||
Acremonium hansfordii | CBS 390.73 | HQ232043 | AB540578 | |||
Acremonium hennebertii | CBS 768.69T | HQ232044 | HF680238 | |||
Acremonium inflatum | CBS 212.69T | HQ232050 | ||||
Acremonium mali | ACCC 39305T | MF993114 | MF987658 | |||
Acremonium moniliforme | CBS 139051T | LN810507 | LN810516 | |||
Acremonium moniliforme | FMR 10363 | LN810508 | LN810517 | |||
Acremonium parvum | CBS 381.70A | HQ231986 | HF680219 | |||
Acremonium persicinum | CBS 310.59T | HQ232077 | ||||
Acremonium persicinum | CBS 101694 | HQ232085 | ||||
Acremonium pinkertoniae | CBS 157.70T | HQ232089 | HQ232202 | |||
Acremonium polychroma | CBS 181.27T | HQ232091 | AB540567 | |||
Acremonium potronii | CBS 189.70 | HQ232094 | ||||
Acremonium pseudozeylanicum | CBS 560.73T | HQ232101 | ||||
Acremonium pteridii | CBS 782.69T | HQ232102 | ||||
Acremonium pteridii | CBS 784.69 | HQ232103 | ||||
Acremonium sclerotigenum | CBS 124.42T | HQ232126 | FN706552 | HQ232209 | ||
Acremonium sclerotigenum | A101 | KC987215 | KC987139 | KC987177 | KC998961 | |
Acremonium sclerotigenum | A130 | KC987242 | KC987166 | KC987204 | KC998988 | |
Acremonium sp. | E102 | KC987248 | KC987172 | KC987210 | KC998994 | KC999030 |
Acremonium spinosum | CBS 136.33T | HQ232137 | HE608637 | HQ232210 | ||
Acremonium stroudii | CBS 138820T | KM225291 | ||||
Acremonium tumulicola | CBS 127532T | AB540478 | AB540552 | |||
Acremonium variecolor | CBS 130360T | HE608651 | HE608647 | |||
Acremonium variecolor | CBS 130361 | HE608652 | HE608648 | |||
Acremonium verruculosum | CBS 989.69T | HQ232150 | ||||
Acrophialophora hechuanensis | GZUIFR-H08-1T | MK926789 | DQ185070 | EU053286 | ||
Brunneomyces brunnescens | CBS 559.73T | HQ231966 | LN810520 | HQ232184 | LN810534 | |
Brunneomyces hominis | UTHSC 06-415T | LN810509 | KP131517 | LN810535 | ||
Bryocentria brongniartii | M139 | EU940105 | EU940052 | |||
Bryocentria brongniartii | M190 | EU940125 | EU940052 | |||
Bryocentria metzgeriae | M140 | EU940106 | ||||
Bulbithecium hyalosporum | CBS 318.91T | AF096187 | HE608634 | |||
Cephalosporium purpurascens | CBS 149.62T | HQ232071 | ||||
Cosmospora lavitskiae | CBS 530.68T | HQ231997 | ||||
Emericellopsis alkalina | CBS 127350T | KC987247 | KC987171 | KC987209 | KC998993 | KC999029 |
Emericellopsis terricola | CBS 120.40T | U57082 | U57676 | U44112 | ||
Gliomastix roseogrisea | CBS 134.56T | HQ232121 | ||||
Hapsidospora irregularis | ATCC 22087T | AF096192 | AF096177 | |||
Kiflimonium curvulum | CBS 430.66T | HQ232026 | HE608638 | HQ232188 | ||
Lanatonectria flavolanata | CBS 230.31 | HQ232157 | ||||
Leucosphaerina arxii | CBS 737.84T | HE608662 | HE608640 | |||
Nigrosabulum globosum | ATCC 22102T | AF096195 | ||||
Paracremonium contagium | CBS 110348T | HQ232118 | KM231831 | KM231966 | ||
Parasarocladium breve | CBS 150.62T | HQ232005 | ||||
Parasarocladium radiatum | CBS 142.62T | HQ232104 | HQ232205 | |||
Pestalotiopsis hawaiiensis | CBS 114491T | KM116239 | KM199339 | KM199514 | ||
Pestalotiopsis spathulata | CBS 356.86T | KM116236 | KM199338 | KM199513 | ||
Phialemonium atrogriseum | CBS 604.67T | HQ231981 | HE610367 | FJ176825 | ||
Pseudoacremonium sacchari | CBS 137990T | KJ869201 | KJ869144 | |||
Sarcopodium vanillae | CBS 100582 | HQ232174 | KM231780 | KM231911 | ||
Sarocladium bacillisporum | CBS 425.67T | HQ231992 | HE608639 | HQ232179 | ||
Sarocladium bactrocephalum | CBS 749.69T | HQ231994 | HG965006 | HQ232180 | ||
Sarocladium strictum | CBS 346.70T | HQ232141 | AY214439 | HQ232211 | ||
Sarocladium terricola | CBS 243.59T | HQ232046 | HQ232196 | |||
Selinia pulchra | AR 2812 | GQ505992 | HM484859 | HM484841 | ||
Trichothecium crotocinigenum | CBS 129.64T | HQ232018 | AJ621773 | |||
Trichothecium indicum | CBS 123.78T | AF096194 | AF096179 | |||
Trichothecium roseum | DAOM 208997 | U69891 | U69892 | |||
Trichothecium sympodiale | ATCC 36477 | U69889 | U69890 | |||
Acremonium curvum |
|
ON041050 | ON041034 | ON876754 | ON494579 | ON494583 |
Acremonium globosisporum |
|
ON041051 | ON041035 | ON876755 | ON494580 | ON494584 |
Acremonium globosisporum | GZUIFR 22.037 | ON041052 | ON041036 | ON876756 | ON494581 | ON494585 |
Acremonium globosisporum | GZUIFR 22.038 | ON041053 | ON041037 | ON876757 | ON494582 | ON494586 |
The ITS and LSU sequences of Acremonium were downloaded from GenBank (Table
Bayesian inference (BI) and maximum likelihood (ML) methods were used in the analysis. For BI analysis was conducted with MrBayes v3.2 (
Based on a BLAST search (https://blast.ncbi.nlm.nih.gov/Blast.cgi) using the LSU sequences, our isolates were identified as belonging to the genus Acremonium. To further determine the phylogenetic position of these strains, we performed a multi-locus phylogenetic analysis. The dataset was composed of LSU (1–430 bp) and ITS (431–1005 bp) gene, comprising a total of 1005 characters (including gaps). The best-fit partition model for ML analysis and BI analysis is shown in Table
The best-fit substitution models are used in multi-locus phylogenetic construction.
LSU | ITS | |
---|---|---|
ML analysis | TN+F+R5 | GTR+F+R4 |
BI analysis | GTR+F+I+G4 | GTR+F+I+G4 |
Yuncheng East Garden Wildlife Zoo, Yuncheng City, Shanxi Province, China N35°6'26", E111°4'24", isolated from green belt soil, July 2021, Yu-Lian Ren (dried holotype culture
Colonies on PDA and OA at 25 °C attaining 11–13 mm and 9–11 mm diam respectively after 7 d, white, flat or raised, velvety to slightly cottony. On MEA at 25 °C, reaching 8–10 mm after 7 d, white to yellowish white, raised, slimy. Hyphae hyaline, septate, sometimes winding and inflate, 1.5–11.0 µm wide. Sporulation abundant. Phialides are mostly borne singly, hyaline, erect to slightly curved, sometimes forming a collarette, 9.0–22.0 µm long, tapering from 1.5–3.5 µm near the base to 0.5–1.5 µm. Conidia cohering in long chains, with minutely truncate ends, up to 27.5 µm long, globose or subglobose, 2.5–4.5 × 2.5–4.5 µm (x– ± SD = 3.4 ± 0.77 × 3.6 ± 0.52, n = 50) diam. Chlamydospores and teleomorph stage were not observed.
globosisporum. A reference to the global conidia.
Yuncheng East Garden Wildlife Zoo, Yuncheng City, Shanxi Province, China N35°6'26", E111°4'24", isolated from green belt soil, July 2021, Yu-Lian Ren, GZUIFR 22.037, ITS, LSU, SSU, TEF 1-α, RPB2 sequences GenBank ON041036, ON041052, ON876756, ON494581, ON494585; GZUIFR 22.038, ITS, LSU, SSU, TEF 1-α, RPB2 sequences GenBank ON041037, ON041053, ON876757, ON494582, ON494586.
Yuncheng City, Shanxi Province, China.
The phylogeny results showed that the
Zhengzhou Zoo, Zhengzhou City, Henan Province, China N34°47'20", E113°40'41", isolated from green belt soil, July 2021, Yu-Lian Ren (dried holotype culture
Colonies on PDA and OA at 25 °C attaining 11–14 mm and 7–9 mm diam respectively after 7 d, white, flat, radially folded or rugose. On MEA at 25 °C, reaching 6–8 mm after 7 d, white to yellowish-white, slimy. Hyphae hyaline, septate, sometimes winding, 1.5–2.5 µm wide. Sporulation abundant. Phialides are mostly borne singly, curved, slightly inflated at the base, tapered at the tip, up to 38.0 µm long. tapering from 1.5–3.5 µm near the base to 0.5–1.5 µm. Conidia cohering together on the top of phialides, one-celled, solitary, or several fascicled, ovoid or subglobose, 3.0–7.0 × 2.5–3.5 µm (x– ± SD = 4.1 ± 1.18 ×3.2 ± 0.77, n = 50) diam. Chlamydospores and teleomorph stage were not observed.
curvum. Referring to the curved Phialides.
Henan Province, China.
Based on the multi-locus analysis we found that Acremonium curvum had close phylogenetic affinities to other taxa of the Chrysogenum-clade. Morphologically, A. curvum was similar to other taxa of the Chrysogenum-clade in having simple or rarely branched conidiophores, slightly inflated at the base and tapered at tip phialides, and ovoid to subglobose conidia (
In the present study, four strains of Acremonium fungi were isolated from soil in the Shanxi and Henan Province, China. Two-locus (LSU and ITS) phylogenetic analyses in combination with morphological characteristics were used for identification. As a result, two new species of A. curvum (one isolate) and A. globosisporum (three isolates) were proposed.
With the development of biotechnology, a growing number of studies have combined morphological and phylogenetic features to distinguish between species. This provides the basis for more precise species naming. Generally, the fungal ITS marker includes considerably more sequence variability, and consequently provides high interspecific resolution, and also some degree of intraspecific variability (
Although,
In recent years, Acremonium spp. has been reported to cause immunocompetent and immunocompromised individual diseases, such as brain abscess (
This work was financially supported by the National Natural Science Foundation of China (no. 32060011, 32160007, 31860002), “Hundred” Talent Projects of Guizhou Province (Qian Ke He [2020] 6005), and the Key Areas of Research and Development Program of Guangdong Province (no. 2018B020205003), and Construction Program of Biology First-class Discipline in Guizhou (GNYL [2017] 009). We appreciate MDPI for the English-language editing of the whole manuscript.