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Research Article
Morphological and phylogenetic analyses reveal two new species in Conidiobolus s.s. (Conidiobolaceae, Entomophthorales) from China
expand article infoYong Nie‡§, Yue Cai|, Heng Zhao, ZhengYu Zhou§, ChangWei Zhao§, XiaoYong Liu#, Bo Huang‡
‡ Anhui Agricultural University, Hefei, China
§ Anhui University of Technology, Ma'anshan, China
| Hefei University, Hefei, China
¶ Beijing Forestry University, Beijing, China
# Shandong Normal University, Jinan, China

Corresponding author: XiaoYong Liu ( 622001@sdnu.edu.cn )

Corresponding author: Bo Huang ( bhuang@ahau.edu.cn )

Academic editor: Kerstin Voigt
© 2023 Yong Nie, Yue Cai, Heng Zhao, ZhengYu Zhou, ChangWei Zhao, XiaoYong Liu, Bo Huang.
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: Nie Y, Cai Y, Zhao H, Zhou Z, Zhao C, Liu X, Huang B (2023) Morphological and phylogenetic analyses reveal two new species in Conidiobolus s.s. (Conidiobolaceae, Entomophthorales) from China. MycoKeys 98: 221-232. https://doi.org/10.3897/mycokeys.98.103603
Open Access

Abstract

The genus Conidiobolus s.s. (Conidiobolaceae, Entomophthorales) has been delimited to accommodate members that produce microspores. Herein, morphological studies, combined with phylogenetic analysis based on the nuclear large subunit of rDNA (nucLSU), the mitochondrial small subunit of rDNA (mtSSU), and the elongation-factor-like gene (EFL) revealed two Conidiobolus s.s. species isolated from plant debris in China. Conidiobolus longiconidiophorus sp. nov. is mainly characterised by its long primary conidiophores, while Conidiobolus polysporus sp. nov. is diagnosed by 2–3 primary conidia arising from branched primary conidiophores. Phylogenetically, the former is grouped into a separate clade, while the latter is closely related to C. incongruus, but is morphologically distinguished by its larger primary conidia and branched conidiophores.

Key words

Conidiobolaceae, microspore, morphology, new taxa, phylogeny

Introduction

The genus Conidiobolus (Ancylistaceae, Entomophthorales) was divided into five genera, i.e. Azygosporus B. Huang & Y. Nie, Capillidium B. Huang & Y. Nie, Conidiobolus s.s. B. Huang & Y. Nie, Microconidiobolus B. Huang & Y. Nie, and Neoconidiobolus B. Huang & Y. Nie based on the molecular and morphological evidences (Nie et al. 2020a; Cai et al. 2021). Subsequently, three families were introduced to accommodate the above five genera based on molecular and genomic data. They were Capillidiaceae Y. Nie, Stajich & K.T. Hodge, Conidiobolaceae B. Huang, Stajich & K.T. Hodge, and Neoconidiobolaceae X.Y. Liu, Stajich & K.T. Hodge (Gryganskyi et al. 2022). The family Conidiobolaceae includes three genera, while Capillidiaceae and Neoconidiobolaceae include one genus each. The genus Conidiobolus s.s. belongs to the family Conidiobolaceae.

Unfortunately, the type species of Conidiobolus, C. utriculosus Brefeld, had been missing for a long time. Therefore, C. coronatus was proposed as the epitype of Conidiobolus s.s. due to its prominence as a pathogenic fungus, its global distribution, and its usage as a model organism for fungal evolution (Spatafora et al. 2016; Möckel et al. 2022). This genus includes 18 species and is the largest among related genera (Goffre et al. 2020; Nie et al. 2020b).

Notably, not all species of Conidiobolus s.s. produce microspores, making it difficult to recognize them without phylogenetic data. These include C. dabieshanensis (Nie et al. 2017), C. iuxtagenitus (Waters and Callaghan 1989), C. margaritatus (Huang et al. 2007), C. taihushanensis (Nie et al. 2020b) and C. lichenicolus (Srinivasan and Thirumalachar 1968). However, their other unique morphological characters and phylogeny could contribute to their suitable identification. Meanwhile, the key to Conidiobolus s.s. was provided to understand the relationship among this fungal group morphologically (Nie et al. 2020b).

This study aims to describe and illustrate two new species of Conidiobolus s.s. based on their morphology and phylogenetic analyses. This study also details the diagnostic characteristics for species that were not observed to produce microspores, and the diversity of Conidiobolus s.s. found in China.

Materials and methods

Isolation and morphology

Plant debris was collected from Guniujiang National Nature Reserve, Qimen County and Shitai County, and Huoli Mountain, Ma,anshan City, Anhui Province, and Yangtianshan National Forest Park, Shandong Province. The strains of Conidiobolus s.s. were isolated from plant debris following the previous described methods (Drechsler 1952; King 1976) and improved by Nie et al. 2012. Plant debris samples were placed into sterilized plastic bags. When they were transferred into the laboratory, the isolation was conducted immediately. Plant debris samples were cut into small pieces with scissors and tiled evenly on the Petri dishes cover, and incubated on inverted Petri dishes containing PDA media (potato 200 g, dextrose 20 g, agar 20 g, H2O 1 L) at 21 °C for 7 days.

The inverted Petri dishes were examined daily by a stereomicroscope (SMZ1500, Nikon Corporation, Japan). When a Conidiobolus-like fungus appeared, it was transferred to a new PDA plate to obtain a pure culture for morphological studies. The micro-morphological structure was observed using a light microscope (BX51, Olympus Corporation, Tokyo, Japan) and imaged using a microscope-camera system (DP25, Olympus Corporation, Tokyo, Japan). The morphological traits of the primary conidia and conidiophores, microconidia, resting spores etc. were described using the method by King (1976). All isolates were deposited at the Engineering Research Center of Biofilm Water Purification and Utilization Technology of Ministry of Education at Anhui University of Technology, Anhui Province, China (BWPU), and duplicated at the Research Center for Entomogenous Fungi at Anhui Agricultural University, Anhui Province, China (RCEF). A total of 14 ex-types of Conidiobolus s.l. were obtained from the American Type Culture Collection, Manassas, VA, USA (ATCC).

DNA extraction, PCR amplification and sequencing

Genomic DNA was extracted from fresh fungal mycelia which were scraped from PDA, using a modified cetyltrimethylammonium bromide (CTAB) protocol as described in Watanabe et al. (2010). Three different loci were amplified using the following primer pairs: LR0R (5’-ACC CGC TGA ACT TAA GC-3’) / LR5 (5’-TCC TGA GGG AAA CTT CG-3’) for nucLSU (Vilgalys and Hester 1990), mtSSU1 (5’-GCW GCA GTG RGG AAT NTT GGR CAA T-3’) / mtSSU2R (5’-GTR GAC TAM TSR GGT ATC TAA TC-3’) for mtSSU (Zoller et al. 1999), and EF983 (5’-GCY CCY GGH CAY CGT GAY TTY AT-3’) / EF1aZ-1R (5’-ACA TCW CCG ACA CCC TTG ATC TTG -3’) for EFL (Nie et al. 2012).

Polymerase chain reaction (PCR) amplification reactions contained 1 μL dNTPs (200 μM), 1 μL MgCl2 (2.5 mM), 10 µL Phusion HF buffer (5x), 1 μL primers each (0.5 μM), 100 ng genomic DNA, and 0.5 μL Taq polymerase (0.04 Unit/L, Super Pfx DNA Polymerase, Cowinbioscience Co. Ltd., Shanghai, China). PCR amplificated program followed Nie et al. (2020b). Bi-directional sequencing was generated by Shanghai Genecore Biotechnologies Company (Shanghai, China). Sequences were processed with Geneious 9.0.2 (http://www.geneious.com, Kearse et al. 2012) to obtain consensus sequences. All sequences were deposited in GenBank (Table 1).

Table 1.
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The species used in phylogenetic analyses.

Species Strains* GenBank accession numbers
nucLSU EFL mtSSU
Azygosporus macropapillatus CGMCC 3.16068 (T) MZ542006 MZ555650 MZ542279
parvus ATCC 14634 (T) KX752051 KY402207 MK301192
Conidiobolus bifurcatus CGMCC 3.15889 (T) MN061285 MN061482 MN061288
C. brefeldianus ARSEF 452 (T) EF392382 – EF392495
C. chlamydosporus ATCC 12242 (T) JF816212 JF816234 MK301178
C. coronatus NRRL 28638 AY546691 DQ275337 –
RCEF 4518 JN131537 JN131543 –
C. dabieshanensis CGMCC 3.15763 (T) KY398125 KY402206 MK301180
C. firmipilleus ARSEF 6384 JX242592 – JX242632
C. gonimodes ATCC 14445 (T) JF816221 JF816226 MK301182
C. humicolus ATCC 28849 (T) JF816220 JF816231 MK301184
C. incongruus NRRL 28636 AF113457 – –
C. iuxtagenitus ARSEF 6378 (T) KC788410 – –
RCEF 4445 JX946695 JX946700 MK333391
C. khandalensis ATCC 15162 (T) KX686994 KY402204 MK301185
C. lichenicolus ATCC 16200 (T) JF816216 JF816232 MK301186
C. longiconidiophorus sp. nov. RCEF 6563 (T) OQ540746 OQ550509 OQ540744
RCEF 6568 (T) OR100884 OR113355 OR100881
C. macrosporus ATCC 16578 (T) KY398124 KY402209 MK301188
C. megalotocus ATCC 28854 (T) MF616383 MF616385 MK301189
C. mycophagus ATCC 16201 (T) JX946694 JX946698 MK301190
C. mycophilus ATCC 16199 (T) KX686995 KY402205 MK301191
C. polyspermus ATCC 14444 (T) MF616382 MF616384 MK301193
C. polysporus sp. nov. RCEF 4500 MG272478 MG272476 OR100882
RCEF 7058 (T) OQ540747 OQ550510 OQ540745
C. polytocus ATCC 12244 (T) JF816213 JF816227 MK301194
C. taihushanensis CGMCC 3.15900 (T) MT250086 MT274290 MT250088
C. variabilis CGMCC 3.15901 (T) MT250085 MT274289 MT250087
Microconidiobolus nodosus ATCC 16577 (T) JF816217 JF816235 MK333388
M. paulus ARSEF 450 (T) KC788409 – –
M. terrestris ATCC 16198 (T) KX752050 KY402208 MK301199

*ARSEF, ARS Entomopathogenic Fungus Collection (Ithaca, U.S.A.). ATCC, American Type Culture Collection (Manassas, U.S.A). CGMCC, China General Microbiological Culture Collection Center (Beijing, China). NRRL, ARS Culture Collection (Peoria, U.S.A). RCEF, Research Center for Entomogenous Fungi (Hefei, China). T = ex-type. The new species reported in this study are indicated in bold.

Phylogenetic analyses

According to our previous studies (Nie et al. 2020a, b), the sequences of three loci (nucLSU, mtSSU, and EFL) of Conidiobolus s.s. species were retrieved from GenBank. Two Azygosporus and two Microconidiobolus species were chosen as out groups. Newly generated sequences from the three strains were aligned with all reference sequences by MAFFT program (Katoh and Standley 2013) and manually corrected with BioEdit (Hall 1999). The final alignments of three loci were concatenated using SequenceMatrix (Vaidya et al. 2011). The output sequence matrix was deposited in TreeBase (https://treebase.org) with the submission ID 30475. Maximum Likelihood (ML) and Bayesian Inference (BI) phylogenetic analyses were conducted. The best-fit substitution model of each partition was evaluated by MrModeltest 2.3 (Nylander 2004). The ML phylogenetic analysis was statistically tested in RAxML 8.1.17 with 1000 bootstrap replicates (Stamatakis 2014). The BI phylogenetic analyses included four MCMC chains and ran for 0.50 million generations until the average standard deviation of split frequencies was below 0.01. The trees were saved once every 100 generations. The burn-in fraction was set to 0.25 and posterior probabilities (PP) were determined from the remaining trees. Phylogenetic trees were checked with FigTree 1.4 (Rambaut 2012) and modified with Adobe Illustrator CS6.0 and Adobe Photoshop CS3.0.

Results

Phylogenetic analyses

The concatenated alignment utilized in this study comprised 1899 characters of nucLSU (1–984), EFL (985–1485), and mtSSU (1486–1899), out of which 986 characters are constant, 289 characters were found to be parsimony-uninformative and 624 characters were parsimony-informative. The best substitution model GTR+I+G was chosen for all the partitions during the ML and BI phylogenetic analyses. The final average standard deviation of the split frequencies was 0.00841, and the BI tree topology was found to be similar to that of ML. Therefore, the best scoring RAxML tree was used to represent the phylogenetic relationships among the studied taxa, with a final likelihood value of -13552.35 (Fig. 1). The phylogeny demonstrated that the three strains RCEF 4500 / 6563 / 6568 / 7058 in the present study were grouped with were members of the genus Conidiobolus s.s, revealing that the three strains belong to the family Conidiobolaceae, the genus Conidiobolus s.s. Furthermore, the strain RCEF 6563 and RCEF 6568 were grouped in an independent clade with a sound support (-/ 0.99), while the strains of RCEF 4500 and RCEF 7058 were claded with C. incongruus with a higher support (83 / 0.98).

Figure 1. 

The phylogenetic tree of Conidiobolus s.s. constructed based on combined nucLSU, EFL and mtSSU sequences. Azygosporus and Microconidiobolus species were used as outgroups. New species are shown in red. Maximum Likelihood bootstrap values (≥70%) / Bayesian posterior probabilities (≥0.95) of clades are provided alongside the branches. The scale bar at the bottom left indicates substitutions per site.

Taxonomy

Conidiobolus longiconidiophorus B. Huang & Y. Nie, sp. nov.

MycoBank No: MycoBank No: MB84768
Fig. 2

Etymology

Longiconidiophorus (Lat.), referring to the long size of its conidiophores.

Known distribution

Anhui Province, China.

Typification

China, Anhui Province, Huangshan City, Qimen County, Guniujiang National Nature Reserve, 30°2′84′′N, 117°53′31′′E, from plant debris, 12 Dec. 2019, Y. Nie and W. Wang, holotype BWPU 191212. Ex-type culture RCEF 6563. GenBank: nucLSU = OQ540746; EFL = OQ550509; mtSSU = OQ540744.

Additional specimens examined

China, Anhui Province, Chizhou City, Shitai County, Guniujiang National Nature Reserve, 30°10’66"N, 117°50’4"E, from plant debris, 15 Dec. 2019, Y. Nie and W. Wang, culture RCEF 6568. GenBank: nucLSU = OR100884; EFL = OR113355; mtSSU = OR100881.

Description

Colonies on PDA at 21 °C after 3 d white, reaching ca 15 mm in diameter. Aerial hyphae flourishing after 6 d. Mycelia white, 5–10 μm wide, often unbranched at the edge of colony. Primary conidiophores often evolving from aerial hyphae, long, 150–340 × 6–9 μm, unbranched and producing a single primary conidium, without widening upward near the tip. Primary conidia forcibly discharged, globose, obovoid to ellipsoidal, 31–49 × 24–42 μm, papilla tapering and pointed, 7–13 μm wide, 3–7 μm long. Secondary conidiophores short or long, arising from primary conidia, bearing a single similar replicative conidium to primary conidia. Microspores not observed on the 2% water agar, but the structure similar to sterigmatas bearing microspores observed. Resting spores absent after 1 month.

Notes

Conidiobolus longiconidiophorus forms a distinct phylogenetic clade from other Conidiobolus s.s. species. Morphologically, its primary condia are similar in size to those in C. coronatus (Cost.) Batko (14.5–38.5 × 17–48.5 μm), C. dabieshanensis Y. Nie & B. Huang (29–38 × 32.5–45), C. macrosporus Srin. & Thirum. (38–45 × 48–54 μm), C. megalotocus Srin. & Thirum. (30–50 μm), and C. utriculosus Brefeld (25–35 × 37.5–51 μm). However, it can be distinguished from C. coronatus and C. macrosporus by its longer primary conidiophores and the absence of resting spores (Batko 1964; Srinivasan and Thirumalachar 1967). Additionally, it is differentiated from C. dabieshanensis and C. utriculosus by its obovoid and ellipsoidal primary condia, as well as the absence of resting spores (Brefeld 1884; Nie et al. 2017). While it is closely related to C. megalotocus, it can be differentiated by the shape of its primary condia (Srinivasan and Thirumalachar 1962). Furthermore, in the phylogenetic tree (Fig. 1), C. longiconidiophorus is found to be distantly related to C. megalotocus.

Figure 2. 

Conidiobolus longiconidiophorus RCEF 6563 a colony on PDA after 3 d at 21 °C b colony on PDA after 6 d at 21 °C c mycelia unbranched at the edge of the colony d–g primary conidiophores bearing primary conidia h, i globose primary conidia j, k obovoid to ellipsoidal primary conidia l, m primary conidia bearing a single secondary conidium n, o structure similar to sterigmatas arsing from conidia. Scale bars: 100 μm (c); 20 μm (d–o).

Conidiobolus polysporus B. Huang & Y. Nie, sp. nov.

MycoBank No: MycoBank No: MB84769
Fig. 3

Etymology

Polysporus (Lat.), referring to several primary conidia arising from branched primary conidiophores.

Known distribution

Anhui and Shandong Provinces, China.

Typification

China, Anhui Province, Ma,anshan City, Huoli Mountain, 31°67′5′′N, 118°55′37′′E, from plant debris, 3 Nov. 2021, Z.Y. Zhou and C.W Zhao, holotype BWPU 211103. Ex-type culture RCEF 7058. GenBank: nucLSU = OQ540747; EFL = OQ550510; mtSSU = OQ540745.

Additional specimens examined

China, Shandong Province, Qingzhou City, Yangtianshan National Forest Park, 36°46’31"N, 118°32’56"E, from plant debris, 18 Mar 2009, C.F. Wang, culture RCEF 4500. GenBank: nucLSU = MG272478; EFL = MG272476; mtSSU = OR100881.

Description

Colonies on PDA at 21 °C after 3 d white, reaching ca 20–23 mm in diameter. Mycelia colorless, rarely branched at the edge of colony, 8.8–13 μm wide, vegetative hyphae filamentous, frequently appearing pronouncedly vacuolated, 15–22 μm wide. Primary conidiophores often unbranched, producing a single primary conidium, without widening upward near the tip, but in some instances bifurcate thus bearing two primary conidia, or forming three conidiophores at the tip thus bearing three primary conidia, 68–270 × 11–19 μm. Primary conidia forcibly discharged, mostly globose, 42–55 × 33–45 μm, Papilla 7.5–14 μm wide, 4–12 μm long. Sometimes obovoid, up to 65 μm long. Secondary conidia arising from primary conidia, similar and smaller to the primary conidia. Microconidia rarely observed on the 2% water agar, globose to elongate ellipsoidal, 7.5–8.8×7.5–12.5 μm. Zygospores formed between adjacent segments after 15 days, smooth, mostly globose, less often ellipsoidal, 17.5–37 μm in diameter, with a 1–3 μm thick wall.

Notes

Conidiobolus polysporus is characterized by several primary conidia (2–3) arising from conidiophores, which are similar to those in C. polytocus Drechsler and C. taihushanensis B. Huang & Y. Nie. However, C. polysporus has larger primary conidia (42–55 × 33–45 μm in C. polysporus vs. 14–29 × 12–25 μm in C. polytocus), and forms zygospores while resting spores are absent in C. polytocus (Drechsler 1955). In addition, C. polysporus differs from C. taihushanensis due to its larger primary conidia (42–55 × 33–45 μm in C. polysporus vs. 27–42 × 19–32 μm in C. taihushanensis) and smaller zygospores (17.5–37 μm in C. polysporus vs. 34–48 × 23–40 μm in C. taihushanensis) (Nie et al. 2020b). Moreover, it is distantly related to C. polytocus and C. taihushanensis in the phylogenetic tree (Fig. 1). Although C. polysporus is grouped with C. incongruus, it can be distinguished by its larger primary conidia (42–55 × 33–45 μm in C. polysporus vs. 18–42 × 13–37 μm in C. incongruus) and branched conidiophore (Drechsler 1960).

Figure 3. 

Conidiobolus polysporus RCEF 7058 a colony on PDA after 3 d at 21 °C b mycelia unbranched at the edge of the colony c, d hyphae appearing pronouncedly vacuolated e, f unbranched primary conidiophores g, h branched primary conidiophpores i–k globose primary conidia l obovoid primary conidia m, n primary conidia bearing a single secondary conidium o–q microconidia arising from a conidium r, s zygospores formed between adjacent segments of the same hypha t, u zygospores. Scale bars: 100 μm (b); 20 μm (c–u).

Discussion

Although the family Conidiobolaceae was originally proposed to include three genera, Azygosporus, Conidiobolus s.s., and Microconidiobolus, recent phylogenetic analyses by Gryganskyi et al. (2022) revealed that the genus Microconidiobolus should be placed in a separate clade (Gryganskyi et al. 2022). In addition, we found that this fungal group produces smaller primary conidia, mostly less than 20 μm in size, without microspores, in comparison to most members of Azygosporus and Conidiobolus s.s. (Nie et al. 2020a). Therefore, it may be appropriate to recognize Microconidiobolus as a distinct family rather than a genus in the family Conidiobolaceae. However, additional evidence, including unique morphological characteristics, phylogenetic analyses with more taxa, and more genome data, is necessary to confirm this hypothesis. C. longiconidiophorus produces long primary conidiophores (over 300 μm) because most of them develop from aerial hyphae. We noticed that C. dabieshanensis (Nie et al. 2017) also produces such long primary conidiophores (up to 287 μm), and they are closely grouped together in the phylogenetic tree (Fig. 1). Coincidentally, these two species were not observed to produce microspores. Nevertheless, we made several attempts, such as culturing at low or high temperatures, on different culture media, and even exposing them to ultraviolet radiation to induce microspore formation. However, we were still unable to observe microspores, and we hypothesized that microspores of these species may only arise under the natural environment. This phenomenon was also observed in four other Conidiobolus s.s. species and may require further investigation.

Conidiobolus polysporus is known to produce 2–3 primary conidia arising from branched primary conidiophores. Similar branched primary conidiophores have also been observed in C. gonimodes (Drechsler 1961), C. margaritatus (Huang et al. 2007), C. polytocus (Drechsler 1955) and C. taihushanensis (Nie et al. 2020b). However, the number of primary conidia borne on these branched primary conidiophores varies: C. gonimodes and C. margaritatus produce 2 primary conidia, C. polytocus produces 2–4 primary conidia, and C. taihushanensis produces 2–6 primary conidia. Notably, the two primary conidia of C. gonimodes arise directly from the top of branched primary conidiophores without short handles (Drechsler 1961). Additionally, C. polysporus produces primary conidia that are larger than those produced by the other four Conidiobolus s.s. species mentioned above.

Interestingly, we found that C. iuxtagenitus was located at the bottom of the phylogenetic tree and was distinct from other Conidobolus s.s. members. C. iuxtagenitus is characterized by an absence of microspore and its zygospores formed by a short beak near a lateral conjugation (Waters and Callaghan 1989). Therefore, it is possible that C. iuxtagenitus represents another potential new lineage.

In this study, we introduce two new species of Conidiobolus s.s. species, namely C. longiconidiophorus and C. polysporus, based on morphological and phylogenetic evidence. These findings expand the number of known Conidiobolus s.s. species to 20.

Acknowledgements

This study was supported by the National Natural Science Foundation of China (Nos. 31900008, 30770008 and 31970009). We also thank the editor and anonymous reviewer for their valuable comments.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This study was supported by the National Natural Science Foundation of China (Nos. 31900008, 30770008 and 31970009).

Author contributions

Writing—original draft preparation, Y.N.; investigation and resources, Y.C.; software, H.Z.; methodology, Z.-Y.Z and C.-W.Z; writing—review and editing, conceptualization and supervision, X.-Y.L and B.H. All authors have read and agreed to the published version of the manuscript.

Author ORCIDs

Yong Nie https://orcid.org/0000-0001-8964-1661

Yue Cai https://orcid.org/0000-0002-4970-9673

Heng Zhao https://orcid.org/0000-0003-2938-5613

ZhengYu Zhou https://orcid.org/0000-0001-9312-1626

ChangWei Zhao https://orcid.org/0009-0007-7573-346X

XiaoYong Liu https://orcid.org/0000-0002-8808-010X

Bo Huang https://orcid.org/0000-0001-6032-7396

Data availability

All of the data that support the findings of this study are available in the main text.

References

  • Batko A (1964) Notes on entomophthoraceous fungi in Poland Entomophaga. Mémoires hors série 2: 129–131.
  • Brefeld O (1884) Conidiobolus utriculosus und minor. Untersuchungen aus der Gesammtgebiete der Mykologie 6(2): 35–78.
  • Goffre D, Jensen AB, Lopez Lastra CC, Humber RA, Folgarait PJ (2020) Conidiobolus lunulus, a new entomophthoralean species isolated from leafcutter ants. Mycologia 113(1): 56–64. https://doi.org/10.1080/00275514.2020.1816387
  • Gryganskyi AP, Nie Y, Hajek AE, Hodge KT, Liu XY, Aadland K, Voigt K, Anishchenko IM, Kutovenko VB, Kava L, Vuek A, Vilgalys R, Huang B, Stajich JE (2022) The early terrestrial fungal lineage of Conidiobolus—Transition from saprotroph to parasitic lifestyle. Journal of Fungi 8(8): e789. https://doi.org/10.3390/jof8080789
  • Hall TA (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98.
  • Huang B, Humber RA, Hodge KT (2007) A new species of Conidiobolus from Great Smoky Mountains National Park. Mycotaxon 100: 227–233.
  • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30(4): 772–780. https://doi.org/10.1093/molbev/mst010
  • Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics (Oxford, England) 28(12): 1647–1649. https://doi.org/10.1093/bioinformatics/bts199
  • King DS (1976) Systematics of Conidiobolus (Entomophthorales) using numerical taxonomy I. Taxonomic considerations. Canadian Journal of Botany 54: 45–65. https://doi.org/10.1139/b76-008
  • Möckel L, Meusemann K, Misof B, Schwartze VU, De Fine Licht HH, Voigt K, Stielow B, de Hoog S, Beutel RG, Buellesbach J (2022) Phylogenetic revision and patterns of host specificity in the fungal Subphylum Entomophthoromycotina. Microorganisms 10(2): e256. https://doi.org/10.3390/microorganisms10020256
  • Nie Y, Yu CZ, Liu XY, Huang B (2012) A new species of Conidiobolus (Ancylistaceae) from Anhui, China. Mycotaxon 120(1): 427–435. https://doi.org/10.5248/120.427
  • Nie Y, Tang XX, Liu XY, Huang B (2017) A new species of Conidiobolus with chlamydospores from Dabie Mountains, eastern China. Mycosphere: Journal of Fungal Biology 8(7): 809–816. https://doi.org/10.5943/mycosphere/8/7/1
  • Nie Y, Yu DS, Wang CF, Liu XY, Huang B (2020a) A taxonomic revision of the genus Conidiobolus (Ancylistaceae, Entomophthorales): Four clades including three new genera. MycoKeys 66: 55–81. https://doi.org/10.3897/mycokeys.66.46575
  • Nie Y, Cai Y, Gao Y, Yu DS, Wang ZM, Liu XY, Huang B (2020b) Three new species of Conidiobolus sensu stricto from plant debris in eastern China. MycoKeys 73: 133–149. https://doi.org/10.3897/mycokeys.73.56905
  • Nylander JAA (2004) MrModeltest v. 2. Program distributed by the author. Evolutionary Biology Centre, Uppsala University.
  • Spatafora JW, Chang Y, Benny GL, Lazarus K, Smith ME, Berbee ML, Bonito G, Corradi N, Grigoriev I, Gryganskyi A, James TY, O’Donnell K, Roberson RW, Taylor TN, Uehling J, Vilgalys R, White MM, Stajich JE (2016) A phylum-level phylogenetic classification of zygomycete fungi based on genome-scale data. Mycologia 108(5): 1028–1046. https://doi.org/10.3852/16-042
  • Srinivasan MC, Thirumalachar MJ (1962) Studies on species of Conidiobolus from India-II. Sydowia. Annales Mycologici 16: 60–66.
  • Srinivasan MC, Thirumalachar MJ (1968) Studies on species of Conidiobolus from IndiaV. Mycopathologia et Mycologia Applicata 36(3–4): 341–346. https://doi.org/10.1007/BF02050380
  • Vaidya G, Lohman DJ, Meier R (2011) SequenceMatrix: Concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics 27(2): 171–180. https://doi.org/10.1111/j.1096-0031.2010.00329.x
  • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172(8): 4238–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990
  • Watanabe M, Lee K, Goto K, Kumagai S, Sugita-Konishi Y, Hara-Kudo Y (2010) Rapid and effective DNA extraction method with bead grinding for a large amount of fungal DNA. Journal of Food Protection 73(6): 1077–1084. https://doi.org/10.4315/0362-028X-73.6.1077
  • Waters SD, Callaghan AA (1989) Conidiobolus iuxtagenitus, a new species with discharge delongate repetitional conidia and conjugation tubes. Mycological Research 93(2): 223–226. https://doi.org/10.1016/S0953-7562(89)80121-2
  • Zoller S, Scheideggera C, Sperisena C (1999) PCR primers for the amplification of mitochondrial small subunit ribosomal DNA of lichen-forming ascomycetes. Lichenologist (London, England) 31(5): 511–516. https://doi.org/10.1006/lich.1999.0220
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