Research Article |
Corresponding author: Yan-Feng Han ( swallow1128@126.com ) Academic editor: Danny Haelewaters
© 2019 Zhi-Yuan Zhang, Wan-Hao Chen, Xiao Zou, Yan-Feng Han, Jian-Zhong Huang, Zong-Qi Liang, Sunil K. Deshmukh.
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:
Zhang Z-Y, Chen W-H, Zou X, Han Y-F, Huang J-Z, Liang Z-Q, Deshmukh SK (2019) Phylogeny and taxonomy of two new Plectosphaerella (Plectosphaerellaceae, Glomerellales) species from China. MycoKeys 57: 47-60. https://doi.org/10.3897/mycokeys.57.36628
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The genus Plectosphaerella is the largest genus in the family Plectosphaerellaceae. Some species are plant pathogens, whereas others are soil-borne. Seven Plectosphaerella isolates were collected from various locations in the southwest of China. Using multi-locus phylogenetic (LSU, ITS, EF1α, RPB2) analyses combined with morphological characteristics, two new species, Plectosphaerella guizhouensis sp. nov. and Plectosphaerella nauculaspora sp. nov. are described, illustrated and compared with related species.
Filamentous fungi, Plectosphaerellaceae, Multi-locus, Morphology, Taxonomy
The genus Plectosphaerella Kleb., established in 1929, is the largest genus in the family Plectosphaerellaceae (Sordariomycetes, Glomerellales) (
Members of the genus Plectosphaerella are isolated from different habitats throughout the world, including plants, animals and soil. For example, P. tabacinum (J.F.H. Beyma) M.E. Palm, W. Gams & Nirenberg (the anamorph of P. cucumerina) has a cosmopolitan distribution with reports in Canada and the USA (North America), Belgium, England, Italy, The Netherlands and Switzerland (Europe), Egypt (Africa) etc. (
During the investigation of keratinolytic fungi from different soils in China, seven isolates in the genus Plectosphaerella were obtained in Guizhou Province, China. The aim of our project was to identify these isolates, based on combined molecular phylogeny and morphological characteristics.
Soil samples were collected from Qianlingshan Park (26°60'N, 106°69'E), Guiyang city and the affiliated hospital of Zunyi Medical University (27°70'N, 106°94'E), Zunyi city, Guizhou Province, China by Zhi-Yuan Zhang on 10 Sept. 2016. Samples were collected 3–10 cm below the soil surface and placed in Ziploc plastic bags. Isolation and purification of strains were undertaken according to methods described by
Type collections of the novel species are deposited in the Mycological Herbarium of the Institute of Microbiology, Chinese Academy of Sciences, Beijing, China (
Total genomic DNA was extracted from fresh fungal mycelia using the BioTeke Fungus Genomic DNA Extraction Kit (DP2032, BioTeke, China), following the manufacturer’s instructions. The internal transcribed spacer (ITS) regions and the 5’ end of the 28S nrRNA locus (LSU) were amplified and sequenced with the primer pairs ITS1/ITS4 (
The DNA sequences, generated in this study, were assembled using Lasergene software (version 6.0, DNASTAR). Sequence data, mostly from
Maximum likelihood (ML) analyses were constructed with IQ-TREE v. 1.6.11 (
For Bayesian Inference (BI), a Markov Chain Monte Carlo (MCMC) algorithm was used to generate phylogenetic trees with Bayesian probabilities using MrBayes v.3.2 (
Fifty-five strains (including the seven with new sequence data) were included in our multi-locus dataset (Table
The analyses of concatenated dataset (Figure
Phylogenetic tree of Plectosphaerella species derived from Bayesian analyses and Maximum Likelihood analyses, based on the combined sequences dataset of LSU+ITS+EF1α+RPB2. Bayesian posterior probabilities (BI pp) greater than 0.7 and Maximum Likelihood bootstrap support values (ML BS) greater than 95% are shown above branches. New isolates are in bold and blue. The tree used Brunneochlamydosporium nepalense (
Referring to Guizhou, the province where the isolate was collected.
Sexual morph not observed. Asexual morph on CA. Mycelium hyaline, smooth, septate, branched and thin-walled, 1–2 μm (x̄=1.5 μm) wide. Conidiophores solitary, unbranched or rarely branched, hyaline, smooth, thin-walled, sometimes radiating out from hyphal coils. Conidiogenous cells growing from a short branch or directly from mycelia, phialides, discrete, polymorphic, cylindrical, sub-cylindrical or ampulliform; terminal or lateral, hyaline, smooth, solitary, straight at the apex, sometimes bent or helicoid, gradually tapering to the apex, 3.5–17 × 0.5–2 μm (x̄ = 9.5 × 1.5 μm, n = 20), collarette cylindrical, 0.5–1 μm deep. Conidia aggregating in slimy heads, non-septate or 1-septate, fusiform or cylindrical, sometimes rounded at both ends, hyaline, smooth, thin-walled; 2–6.5 × 1.5–5 μm (x̄ = 5.5 × 2 μm, n = 10) (1-septate), 3–5 × 1–1.5 μm (x̄ = 4 × 1.5 μm, n = 10) (non-septate). Chlamydospores absent.
Colonies on PDA reaching 74–75 mm diam. in 14 d at 25 °C, milk white, flat, aerial hyphae sparse, floccose at periphery, sub-rounded, margin regular, reverse milk white. Colonies on CA reaching 65–67 mm diam. in 14 d at 25 °C, white to milk white, flat, floccose, margin weakly undulate to faintly fimbriate, reverse milk white.
CHINA, Guizhou, Guiyang, Qianlingshan Park, 26°60'N, 106°69'E, 1210 m a.s.l., on soil, 10 Sep. 2016, collected and isolated by Zhi-Yuan Zhang,
Based on multi-locus phylogenetic analyses (Figure
From “naucula”, referring to the navicular conidia.
Sexual morph not observed. Asexual morph on CA. Mycelium hyaline, smooth, septate, branched and thin-walled, 1–1.2 μm (x̄=1.5 μm) wide. Conidiophores solitary, unbranched or rarely branched, hyaline, smooth, thin-walled, hyphal coils not observed. Conidiogenous cells growing from short branch or directly from mycelia, phialides, discrete, polymorphic, cylindrical, sub-cylindrical or ampulliform; terminal or lateral, hyaline, smooth, gradually tapering to the apex, straight at the apex, sometimes bent or helicoid, 3–37 × 0.5–2 μm (x̄ = 11 × 1 μm, n = 10), collarette minute, cylindrical, 0.5–1 μm deep. Conidia aggregating in slimy heads, 1- or 2–celled, mostly navicular, rarely fusiform or cylindrical, sometimes swollen at both ends, hyaline, smooth, thin-walled, 4–7 × 1–2 μm (x̄ = 5 × 1.5 μm, n = 10) (1-septate), 3–5 × 1–1.5 μm (x̄ = 4 × 1.5 μm, n = 6) (non-septate). Chlamydospores not observed.
Colonies on PDA reaching 75–76 mm diam. in 14 d at 25 °C, milk white, flat, sub-rounded, margin regular, reverse milk white. Colonies on CA reaching 63–65 mm diam. in 14 d at 25 °C, milk white, aerial hyphae sparse, flat, margin weakly undulate to faintly fimbriate, reverse milk white.
CHINA, Guizhou, Guiyang, Qianlingshan Park, 26°60'N, 106°69'E, 1220 m a.s.l., on soil, 10 Sep. 2016, collected and isolated by Zhi-Yuan Zhang,
Phylogenetically, our two isolates
In the present study, seven strains of Plectosphaerella fungi were isolated from soil in the Guizhou Province, China. Multi-locus phylogenetic analyses in combination with morphological data were used for identification. Our study resulted in the description of two new species, P. guizhouensis (3 isolates) and P. nauculaspora (2 isolates). In addition, our two isolates
Plectosphaerella spp. have diverse life styles and habitat sources – including pathogens of several plants, endophytes of plants, pathogens of animals (mainly involving Austropotamobius pallipes and Oratosquilla oratoria) and saprophytes on soil (
At present, more and more studies use combined data from morphological characteristics and molecular phylogeny for identifying new species (e.g.
1 | Growing on crustaceans | P. oratosquillae |
– | On other substrates | 2 |
2 | Teleomorph known | 3 |
– | Teleomorph unknown | 5 |
3 | Ascomata globose or subglobose to pyriform | 4 |
– | Ascomata subglobose to ovoid, or obpyriform | P. kunmingensis |
4 | Asci 50–80 × 6–9 μm | P. cucumerina |
– | Asci 31.4–43 × 6.2–8.2 μm | P. plurivora |
5 | Chlamydospores present | 6 |
– | Chlamydospores absent | 8 |
6 | Conidia mostly septate | 7 |
– | Conidia mostly aseptate | P. melonis |
7 | Conidia 13–19.5 × 2.5–3 μm | P. alismatis |
– | Conidia 6–10 × 1.5–4 μm | P. sinensis |
8 | Phialides branched at tip | 9 |
– | Phialides not branched at tip | 11 |
9 | Phialides 0–3-septate | P. ramiseptata |
– | Phialides 0–1-septate | 10 |
10 | Oligotrophic, polyphialides infrequently seen, collarette 1–2.5 μm | P. oligotrophica |
– | Non-oligotrophic, polyphialides frequently seen, collarette minute | P. pauciseptata |
11 | Conidia ellipsoidal | 12 |
– | Conidia cylindrical, ellipsoidal, fusiform, navicular | 14 |
12 | Conidia mostly septate | P. delsorboi |
– | Conidia aseptate | 13 |
13 | Conidia av. 4 × 2 μm | P. populi |
– | Conidia av. 7.9 × 3.5 μm | P. citrullae |
14 | Conidia mostly navicular | P. nauculaspora |
– | Conidia mostly cylindrical or fusiform | 15 |
15 | Septate conidia 2–6.5 × 1.5–5 μm, aseptate conidia 3–5 × 1–1.5 μm | P. guizhouensis |
– | Septate conidia 7.5–11 × 2.5–3.5 μm, aseptate conidia 5–8 × 2.1–3.3 μm | P. humicola |
We are grateful to the editor Danny Haelewaters (Purdue University, Indiana, USA) and the reviewers – Martina Réblová (Czech Academy of Sciences, Průhonice, Czech Republic) and Yong-Chun Niu (Chinese Academy of Agricultural Sciences, Beijing, China) for comments on the manuscript. This work was financially supported by the Ministry of Science and Technology of China (2013FY110400), Guangdong Technological Innovation Strategy of Special Funds (key areas of research and development programme) (2018B020205003), the National Natural Science Foundation of China (31460010, 31860002) and Construction Program of Biology First-class Discipline in Guizhou (GNYL[2017]009).