Research Article
Print
Research Article
Phylogenetic evidence reveal a close relationship between Amphichorda and Ovicillium in Bionectriaceae (Hypocreales)
expand article infoYao Wang‡, De-Xiang Tang‡, Hui Chen‡, Qi-Rui Li‡, Chanhom Loinheuang§, Xiang-Chun Shen|
‡ Guizhou Medical University, Guizhou, China
§ National University of Laos, Vientiane, Laos
| Guizhou Medical University, Guiyang, China

Corresponding author: Yao Wang ( wangyao1@aliyun.com )

Corresponding author: Xiang-Chun Shen ( shenxiangchun@126.com )

Academic editor: Danushka Sandaruwan Tennakoon
© 2025 Yao Wang, De-Xiang Tang, Hui Chen, Qi-Rui Li, Chanhom Loinheuang, Xiang-Chun Shen.
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: Wang Y, Tang D-X, Chen H, Li Q-R, Loinheuang C, Shen X-C (2025) Phylogenetic evidence reveal a close relationship between Amphichorda and Ovicillium in Bionectriaceae (Hypocreales). MycoKeys 117: 337-352. https://doi.org/10.3897/mycokeys.117.151366
Open Access

Abstract

Animal excrement serves as the primary substrate for Amphichorda, which is found in a wide range of habitats. Based on evolutionary relationships, the genus is currently classified within the Bionectriaceae. However, the phylogenetic position of Amphichorda and its associated taxa remains unresolved due to limited sampling in previous studies. Here, we discovered and identified five Amphichorda species, significantly advancing our understanding of this genus. Using six genomic loci (ITS, nrSSU, nrLSU, tef1α, rpb1, and rpb2) to expand taxonomic sampling, we reconstructed a phylogenetic framework for the Bionectriaceae, with a focus on Amphichorda and related taxa. Phylogenetic analyses revealed a close genetic connection between Amphichorda and related genera, yet they formed distinct clades within the Bionectriaceae and were clearly differentiated. The extensive sampling demonstrated stable phylogenetic relationships among Amphichorda, Hapsidospora, Ovicillium, Proxiovicillium, and Bulbithecium. Furthermore, we described two new species, A. guizhouensis sp. nov. and O. pseudoattenuatum sp. nov., supported by DNA data and morphological characteristics. A comprehensive comparison of morphological traits across all members of Amphichorda and Ovicillium was conducted. This study clarifies taxonomic boundaries and evolutionary relationships within the two genera and contributes to the overall understanding of the biodiversity and systematics of the Bionectriaceae.

Key words:

Coprophilous fungi, morphology, multi-locus phylogeny, new taxa, soil fungi, taxonomy

Introduction

Renowned researcher E.M. Fries created the genus Amphichorda in 1825, designating Amphichorda felina (DC.) Fr. as the type species. The fungus, isolated from cat feces, was characterized by filiform conidiogenous cells and colonies with a white farinaceous hue (Fries 1825). Subsequently, Fries (1832) transferred A. felina to the genus Isaria Pers., and de Hoog (1972) formally designated Isaria felina (DC.) Fr. as the lectotype of the genus, following von Arx’s criteria derived from synnemata production. Based on the morphological similarity of the holoblastic conidiogenous cells, I. felina was accommodated in the genus Beauveria Vuill. as B. felina (DC.) J.W. Carmich. (Carmichael et al. 1980). Aside from its normal conidiogenous cells without an elongated denticulate rachis, Amphichorda shares a morphological resemblance with Beauveria (Rehner et al. 2011; Chen et al. 2013). However, phylogenetic analyses in previous studies revealed significant differences between B. felina and other Beauveria species (Hodge et al. 2005). Therefore, the genus Amphichorda has been reestablished with B. felina based on the phylogenetic relationships of internal transcribed spacer (ITS) sequences and morphology (Zhang et al. 2017). A phylogenetic analysis revealed that Amphichorda species were grouped in a separate clade from Beauveria sensu stricto, indicating the significant phylogenetic separation between the two taxa (Zhang et al. 2017). Further studies by Zhang et al. (2017, 2020) and Liu et al. (2023) placed Amphichorda within Cordycipitaceae and described three new species in the genus. Through a phylogenetic analysis based on a concatenated alignment of the ITS and nuclear ribosomal large subunit (nr LSU) sequences of Bionectriaceae and Cordycipitaceae, Guerra-Mateo et al. (2023) identified Amphichorda as a member of the Bionectriaceae family and established its close phylogenetic link to Hapsidospora Malloch & Cain and Nigrosabulum Malloch & Cain. Hou et al. (2023) determined that Nigrosabulum and Hapsidospora are congeneric, leading to their synonymization within Bionectriaceae based on both phylogeny and morphology. Leão et al. (2024) and Wang et al. (2024) also supported the placement of Amphichorda in the Bionectriaceae family. Despite these numerous studies, the evolutionary connections between Amphichorda and other genera remain unclear. It is imperative to reconstruct the phylogenetic framework for the Bionectriaceae focusing on Amphichorda through more sampling.

Animal excrement serves as the primary substrate for Amphichorda, which are found in a wide range of habitats. Currently, nine Amphichorda species viz. A. cavernicola, A. coprophila, A. excrementa, A. felina, A. guana, A. kunmingensis, A. littoralis, A. monjolensis, and A. yunnanensis have been published in reputable mycological journals (Zhang et al. 2017, 2020; Liu et al. 2023; Guerra-Mateo et al. 2023; Leão et al. 2024; Wang et al. 2024). The majority of the species are coprophilous (such as A. coprophila, A. excrementa, A. guana, and A. kunmingensis); some species are both coprophilous and entomopathogenic (such as A. cavernicola and A. felina); and one species was isolated from marine sediments (A. littoralis), other species from the wing surfaces of Rhinolophus affinis or Rhinolophus siamensis (A. yunnanensis), or a potato dextrose agar plate partially consumed by an insect (A. monjolensis) (Zhang et al. 2017, 2020; Liu et al. 2023; Guerra-Mateo et al. 2023; Leão et al. 2024; Wang et al. 2024).

During an extensive mycological survey conducted in two distinct biogeographical regions spanning China and Laos, seven fungal species were isolated from diverse ecological niches, including soil substrates and animal fecal matter. To complement field-collected specimens, reference strains from the Westerdijk Fungal Biodiversity Institute’s CBS culture collection were incorporated into the study. A multilocus phylogenetic reconstruction integrating ITS, the nuclear ribosomal small subunit (nrSSU), nrLSU, the translation elongation factor 1α (tef1α), the largest subunit of RNA polymerase II (rpb1), and the second subunit of RNA polymerase II (rpb2) sequences confirmed their taxonomic placement within Bionectriaceae. Among these taxa, two novel species, one belonging to the genus Amphichorda and the other to Ovicillium Zare & W. Gams, were proposed and described based on morphological traits and multi-locus molecular phylogenetic data. The morphological features of every component of Amphichorda and Ovicillium were also compared in detail. In addition to introducing and characterizing these two new species, the study aimed to: (1) re-evaluate the taxonomic stability of Amphichorda among related genera within Bionectriaceae, (2) delineate the taxonomic boundaries and evolutionary relationships within Amphichorda and Ovicillium through comparative morphological analysis, and (3) enhance the phylogenetic resolution within Bionectriaceae using six genomic loci.

Materials and methods

Isolates

The samples were collected from three locations: Anshun City, China; Vientiane City, Laos; and Muang Xay District, Oudomxay Province, Laos. The techniques outlined in Wang et al. (2023) were used to isolate the strains from the soil and animal feces. Briefly, 2 g of soil was added to a flask containing 20 mL sterilized water and glass beads. The soil suspension was shaken for about 10 min and then diluted 100 times. Subsequently, 200 µL of the diluted soil suspension was spread on Petri dishes with solidified onion garlic agar (OGA: 20 g of grated garlic and 20 g of onion were boiled in 1 L of distilled water for 1 h; the boiled residue was then filtered off, and 2% (w/v) agar was added to the filtrate). Czapek yeast extract agar (CYA, Advanced Technology and Industrial Co., Ltd., China) and potato dextrose agar (PDA, Difco, USA) were used, and all media had 50 mg/L rose Bengal and 100 mg/L kanamycin added. Conidia developing on animal feces were transplanted onto plates of PDA and cultured at 25 °C. Colonies of the isolated filamentous fungi appearing in the culture were transferred onto fresh PDA media. The purified fungal strain was transferred to PDA slants and cultured at 25 °C until its hyphae spread across the entire slope. The emerging fungal spores were washed with sterile physiological saline and made into a spore suspension of 1 × 103 cells/mL. To obtain monospore cultures, a part of the spore suspension was placed on PDA using a sterile micropipette, and then a Petri dish was incubated at 25 °C. Specimens and type material were deposited in the Guizhou Medical University Herbarium (GMB), China. Living cultures were deposited at the Guizhou Medical University Culture Collection (GMBC). This investigation comprised a total of nine isolates, including species that were tentatively classified as belonging to allied genera, Amphichorda, and Ovicillium (Table 1). Cultures of four Amphichorda species were acquired from the CBS-KNAW Fungal Biodiversity Centre (CBS, Westerdijk Fungal Biodiversity Institute, WI, Utrecht, the Netherlands). Following transfer to PDA medium, the CBS strains were re-cultured.

Table 1.
Download as
CSV
XLSX

Species information and corresponding GenBank accession numbers of Amphichorda and close relative genera used in this study.

Species Strain Genbank accession number Reference
ITS nr SSU nr LSU tef1α rpb1 rpb2
Acremonium acutatum CBS 682.71T OQ429438 N/A OQ055349 OQ470735 N/A OQ453833 Hou et al. 2023
Acremonium alternatum CBS 407.66T OQ429442 N/A OQ055353 OQ470739 N/A OQ560696 Hou et al. 2023
Acremonium chlamydosporium CBS 414.76T OQ429450 N/A OQ055361 OQ470748 N/A OQ453844 Hou et al. 2023
Acremonium cf. egyptiacum CBS 270.86 OQ429463 N/A OQ055374 OQ470760 N/A OQ453857 Hou et al. 2023
Acremonium egyptiacum CBS 114785T OQ429456 N/A OQ055362 OQ470749 N/A OQ453845 Hou et al. 2023
Alloacremonium ferrugineum CBS 102877T OQ429495 N/A OQ055406 OQ470785 N/A OQ453887 Hou et al. 2023
Alloacremonium humicola CBS 613.82T OQ429496 N/A OQ055407 OQ470786 N/A OQ453888 Hou et al. 2023
Amphichorda cavernicola CGMCC 3.19571T MK329056 N/A MK328961 MK335997 N/A N/A Zhang et al. 2020
Amphichorda cavernicola LC12481 MK329057 N/A MK328962 MK335998 N/A N/A Zhang et al. 2020
Amphichorda cavernicola LC12560 MK329061 N/A MK328966 MK336002 N/A N/A Zhang et al. 2020
Amphichorda coprophila CBS 173.71 PQ726811 PQ726824 PQ726836 PQ758601 N/A PQ779067 This study
Amphichorda coprophila CBS 247.82T MH861494 N/A MH873238 OQ954487 N/A N/A Guerra-Mateo et al. 2023
Amphichorda coprophila CBS 424.88 OQ942929 N/A OQ943166 OQ954488 N/A N/A Guerra-Mateo et al. 2023
Amphichorda excrementa YFCC AECCS848T N/A OR913433 OR913439 OR917446 OR917451 OR917443 Wang et al. 2024
Amphichorda excrementa CBS 110.08 PQ726812 PQ726825 PQ726837 PQ758602 PQ758614 PQ779068 This study
Amphichorda feline CBS 250.34 PQ726813 PQ726826 PQ726838 PQ758603 PQ758615 PQ779069 This study
Amphichorda feline CBS 648.66 OQ942930 N/A MH870575 OQ954491 N/A N/A Guerra-Mateo et al. 2023
Amphichorda guana CGMCC3.17908T KU746665 KY883262 KU746711 KX855211 KY883202 KY883228 Zhang et al. 2017
Amphichorda guana CGMCC3.17909 KU746666 KY883263 KU746712 KX855212 KY883203 N/A Zhang et al. 2017
Amphichorda guizhouensis GMBC 3005T PQ726815 PQ726828 PQ726840 PQ758605 PQ758617 PQ779071 This study
Amphichorda guizhouensis GMBC 3006 PQ726816 PQ726829 PQ726841 PQ758606 PQ758618 PQ779072 This study
Amphichorda kunmingensis YFCC AKYYH8414T N/A OR913435 OR913438 OR917448 OR917452 N/A Wang et al. 2024
Amphichorda kunmingensis CBS 312.50 PQ726814 PQ726827 PQ726839 PQ758604 PQ758616 PQ779070 This study
Amphichorda littoralis FMR 17952 OQ942925 N/A OQ943162 OQ954483 N/A N/A Guerra-Mateo et al. 2023
Amphichorda littoralis FMR 19404T OQ942924 N/A OQ943161 OQ954482 N/A N/A Guerra-Mateo et al. 2023
Amphichorda littoralis FMR 19611 OQ942926 N/A OQ943163 OQ954484 N/A N/A Guerra-Mateo et al. 2023
Amphichorda monjolensis COAD 3124T OQ288256 N/A OQ288260 OR454090 N/A OQ405040 Leão et al. 2024
Amphichorda monjolensis COAD 3125 OQ288257 N/A N/A N/A N/A OQ405041 Leão et al. 2024
Amphichorda monjolensis COAD 3120 OQ288258 N/A N/A N/A N/A OQ405042 Leão et al. 2024
Amphichorda yunnanensis KUMCC 21-0414 ON426823 N/A N/A OR025977 OR022016 OR022041 Liu et al. 2023
Amphichorda yunnanensis KUMCC 21-0415 ON426824 N/A N/A OR025976 OR022015 OR022040 Liu et al. 2023
Amphichorda yunnanensis KUMCC 21-0416T ON426825 N/A N/A OR025975 OR022014 OR022039 Liu et al. 2023
Bulbithecium ammophilae CBS 178.78T OQ429504 N/A OQ055415 OQ470793 N/A OQ453895 Hou et al. 2023
Bulbithecium arxii CBS 737.84T OQ429505 N/A OQ055416 OQ470794 N/A OQ451834 Hou et al. 2023
Bulbithecium borodinense CBS 101148T OQ429506 N/A OQ055417 OQ470795 N/A Hou et al. 2023
Bulbithecium ellipsoideum CBS 993.69T OQ429507 N/A OQ055418 OQ470796 N/A OQ453896 Hou et al. 2023
Bulbithecium hyalosporum CBS 318.91T OQ429508 AF096172 OQ055419 OQ470797 N/A OQ453897 Hou et al. 2023
Bulbithecium pinkertoniae CBS 157.70T OQ429509 HQ232202 OQ055420 OQ470799 N/A OQ453898 Hou et al. 2023
Bulbithecium spinosum CBS 136.33T OQ429512 HQ232210 OQ055423 OQ470802 N/A OQ453899 Hou et al. 2023
Bulbithecium truncatum CBS 113718T OQ429513 N/A OQ055424 OQ470803 N/A OQ453900 Hou et al. 2023
Claviceps paspali ATCC 13892 JN049818 U32401 U47826 DQ522321 DQ522367 DQ522416 Spatafora et al. 2007
Claviceps purpurea SA cp11 N/A EF469122 EF469075 EF469058 EF469087 EF469105 Sung et al. 2007
Clonostachys kunmingensis GMBC 3002 PQ726821 PQ726833 PQ726846 PQ758611 N/A PQ758622 This study
Clonostachys rosea GMBC 3003 PQ726822 PQ726834 PQ726847 PQ758612 PQ779065 PQ779076 This study
Clonostachys solani GMBC 3004 PQ726823 PQ726835 PQ726848 PQ758613 PQ779066 PQ779077 This study
Geosmithia lavendula CBS 344.48T OQ429598 N/A OQ055508 OQ470908 N/A OQ453997 Hou et al. 2023
Geosmithia pallidum CBS 260.33T OQ429599 N/A OQ055509 OQ470909 N/A OQ453998 Hou et al. 2023
Hapsidospora chrysogena CBS 144.62T OQ429645 HQ232187 OQ055551 OQ470953 N/A OQ454043 Hou et al. 2023
Hapsidospora flava CBS 596.70T OQ429649 HQ232191 OQ055555 OQ470957 N/A OQ454047 Hou et al. 2023
Hapsidospora globosa CBS 512.70T OQ429655 N/A OQ055561 OQ470963 N/A OQ454053 Hou et al. 2023
Hapsidospora inversa CBS 517.70T OQ429659 N/A OQ055565 OQ470967 N/A OQ454057 Hou et al. 2023
Hapsidospora irregularis CBS 510.70T OQ429660 N/A OQ055566 OQ470968 N/A OQ454058 Hou et al. 2023
Hapsidospora stercoraria CBS 516.70T OQ429662 N/A OQ055568 OQ470970 N/A OQ454060 Hou et al. 2023
Hapsidospora variabilis CBS 100549T OQ429663 N/A OQ055569 OQ470971 N/A OQ454061 Hou et al. 2023
Ovicillium asperulatum CBS 426.95 KU382192 N/A KU382233 OQ471081 N/A OQ454166 Hou et al. 2023
Ovicillium asperulatum CBS 130362T OQ429756 N/A OQ055655 OQ471082 N/A OQ454167 Hou et al. 2023
Ovicillium attenuatum CBS 399.86T OQ429757 N/A OQ055656 OQ471083 N/A OQ454168 Hou et al. 2023
Ovicillium oosporum CBS 110151T OQ429758 N/A OQ055657 OQ471084 N/A OQ454169 Hou et al. 2023
Ovicillium pseudoattenuatum GMBC 3007T PQ726817 PQ726830 PQ726842 PQ758607 PQ779063 PQ779073 This study
Ovicillium pseudoattenuatum GMBC 3008 PQ726818 PQ726831 PQ726843 PQ758608 PQ779064 PQ779074 This study
Ovicillium sinense SD09701T PP836762 N/A PP836764 PP852887 N/A N/A Chen et al. 2024
Ovicillium sinense SD09702 PP836763 N/A PP836765 PP852888 N/A N/A Chen et al. 2024
Ovicillium subglobosum CBS 101963T OQ429759 N/A OQ055658 OQ471085 N/A OQ454170 Hou et al. 2023
Ovicillium variecolor CBS 130360T OQ429760 N/A OQ055659 OQ471086 N/A OQ454171 Hou et al. 2023
Proxiovicillium blochii CBS 324.33 OQ429815 N/A OQ430078 OQ471143 N/A OQ454212 Hou et al. 2023
Proxiovicillium blochii CBS 427.93T OQ429816 HQ232182 OQ430079 OQ471144 N/A OQ454213 Hou et al. 2023
Proxiovicillium lepidopterorum CBS 101239T OQ429817 N/A OQ430080 OQ471145 N/A OQ454214 Hou et al. 2023
Proliferophialis apiculata CBS 303.64T OQ429796 N/A OQ055692 OQ471122 N/A OQ454207 Hou et al. 2023
Proliferophialis apiculata CBS 397.78 OQ429798 N/A OQ055694 OQ471124 N/A OQ454209 Hou et al. 2023
Proliferophialis apiculata CBS 542.79 OQ429799 N/A OQ055695 OQ471125 N/A OQ454210 Hou et al. 2023
Sesquicillium buxi GMBC 3000 PQ726819 N/A PQ726844 PQ758609 PQ758619 PQ758621 This study
Sesquicillium candelabrum GMBC 3001 PQ726820 PQ726832 PQ726845 PQ758610 PQ758620 PQ779075 This study
Stilbocrea colubrensis CBS 141857T MN497406 N/A MN497409 N/A N/A N/A Lechat and Fournier 2019
Stilbocrea macrostoma CBS 141849 OQ429874 N/A OQ430123 OQ471206 N/A OQ454273 Hou et al. 2023
Stilbocrea walteri CBS 144627T OR050519 N/A OQ430124 MH562714 N/A MH577042 Voglmayr and Jaklitsch 2019
Waltergamsia moroccensis CBS 512.82T OQ429943 N/A OQ430193 OQ471276 N/A OQ454343 Hou et al. 2023
Waltergamsia parva CBS 381.70AT OQ429946 N/A OQ430196 OQ471279 N/A OQ454346 Hou et al. 2023
Waltergamsia pilosa CBS 124.70T OQ429949 N/A OQ430199 OQ471282 N/A OQ454349 Hou et al. 2023

Morphological observations

After seven days (for Ovicillium species) or thirty days (for Amphichorda species) in an incubator set at 25 °C, colonies on potato dextrose agar (PDA) were macroscopically described. Characteristics and colony diameters were measured after 7 or 30 days. To take pictures of the colony characters (upper surface and reverse), a Canon 750 D camera (Canon Inc., Tokyo, Japan) was used. Colonies with micromorphological characteristics were seen at 25 °C on PDA. A light microscope (Olympus BX53) was used to examine the micro-morphological features (Conidiophores, Phialides, and Conidia) using sterile water or clear lactophenol cotton blue solution as the mounting medium.

DNA extraction, amplification and sequencing

Axenic cultures grown on PDA plates for 14 days were used for DNA extraction. The CTAB approach, as outlined by Liu et al. (2001), was used to extract genomic DNA. The ITS, nrSSU, nrLSU, tef1α, rpb1, and rpb2 were amplified. For the PCR amplification of six genes, the following primer pairs were employed: The ITS region was amplified using the primer combination ITS5/ITS4 (White et al. 1990). The primer pairs NS1/NS4 and LR0R/LR5 were amplified for the nrSSU region and the nrLSU region (Vilgalys and Hester 1990; White et al. 1990; Hopple 1994); the primer pairs 2218R/983F, RPB1‐5′F/RPB1‐5′R and RPB2-5′F/RPB2-5′R were amplified for the tef1α region, the rpb1 region and the rpb2 region (Rehner and Buckley 2005; Bischoff et al. 2006; Sung et al. 2007). The polymerase chain reaction (PCR) matrix was conducted in a final volume of 50 µl and all detailed information (volume, procedures, etc.) was described by Wang et al. (2023). The consensus sequences were generated by aligning forward and reverse sequencing reads with Geneious Prime 2022 (Biomatters Inc., New Zealand).

Phylogenetic analyses

GenBank provided newly generated sequencing data (http://blast.ncbi.nlm.nih.gov/ (accessed on 20 February 2025)). The sequences mainly referred to recent articles, such as Zhang et al. (2020), Guerra-Mateo et al. (2023) and Hou et al. (2023). MAFFT v. 7 (https://mafft.cbrc.jp/alignment/server/ (accessed on 20 February 2025)) was used to create the sequence alignments for the six distinct loci (ITS, nrSSU, nrLSU, tef1α, rpb1, and rpb2). Where required, the aligned sequences were subsequently manually adjusted. The concatenated alignments (nrSSU + ITS + nrLSU + tef1α + rpb1 + rpb2) were subjected to phylogenetic inferences using the maximum-likelihood (ML) and the Bayesian inference (BI) methods. IQ-tree v.2.1.3 and RAxML7.0.3 were used to conduct ML analyses using 1000 bootstrap replicates and the default general time reversible (GTR) substitution matrix (Stamatakis et al. 2008; Minh et al. 2020). ModelFinder was used to estimate the best evolutionary model for machine learning analyses (Kalyaanamoorthy et al. 2017). The TN+F+I+G4 model was selected as the optimal model for the ML analyses, with 5000 ultrafast bootstraps in a single run (Hoang et al. 2017). jModeltest v. 2.1.4 was used to estimate the optimal substitution model for Bayesian analysis and then performed using MrBayes v. 3.2.6 to assess posterior probabilities (PP) by Markov Chain Monte Carlo sampling (MCMC). For all loci in Bayesian analysis, the GTR+I+G model was suggested based on the results of jModeltest (Darriba et al. 2012; Ronquist et al. 2012). For 3,000,000 generations, four Markov chains ran simultaneously, sampling trees every 1000th generations or until the average standard deviation of split frequencies fell below 0.01 to trigger an automated stop. Following the first 25% of trees being removed as part of the burn-in phase, the posterior probabilities (PP) were computed from the remaining trees. The generated trees were annotated in Microsoft PowerPoint and displayed in FigTree v. 1.4.2 (http://tree.bio.ed.ac.uk/software/figtree (accessed on 20 February 2025)). Each node of the tree displays the ML bootstrap support values (BS) greater than or equal to 80% and the relevant Bayesian posterior probability (PP) more than or equal to 0.90.

Results

Molecular phylogeny

Relevant sequences of 65 strains from GenBank were used in the phylogenetic analyses. The aim was to estimate the phylogeny of Amphichorda and its closely related taxa within the Bionectriaceae family. Claviceps paspali (ATCC 13892) and Claviceps purpurea (SA cp11) were designated as outgroup taxa for the analyses. Six concatenated loci (nrSSU + ITS + nrLSU + tef1α + rpb1 + rpb2) were utilized to analyze the aligned DNA sequence data of 78 strains, as presented in Table 1. The final dataset consisted of 6,509 bp of sequence data, including gaps (nrSSU, 2,012 bp; ITS, 705 bp; nrLSU, 906 bp; tef1α, 995 bp; rpb1, 756 bp; and rpb2, 1,135 bp). Overall, the maximum likelihood (ML) trees generated by IQ-TREE and RAxML exhibited the same clades and tree structure as those obtained from the Bayesian phylogenetic analysis. With the bootstrap support values of the ML analysis (IQ-TREE-BS/RAxML-BS) and pertinent Bayesian posterior probabilities (PP) displayed at the nodes (Fig. 1), the optimal IQ-TREE tree based on the combined dataset was displayed here.

Figure 1. 

Phylogenetic relationships of Amphichorda and related genera in the Bionectriaceae based on combined partial nrSSU + ITS + nrLSU + tef1α + rpb1 + rpb2 sequences. Numbers at the nodes are presented here with ML bootstrap support values (BS) (IQ-TREE-BSIQ > 80%/RAxML-BSRAx > 80%) and relevant Bayesian posterior probabilities (PP) (PP > 0.90). Strains in bold type are those analyzed in this study.

Fourteen well-supported clades were recognized based on both ML and BI analyses of the 78 taxa from Bionectriaceae and Claviceps (Clavicipitaceae, Hypocreales) that accommodate species of the genera Acremonium, Alloacremonium, Amphichorda, Bulbithecium, Clonostachys, Geosmithia, Hapsidospora, Ovicillium, Proxiovicillium, Proliferophialis, Sesquicillium, Stilbocrea, Waltergamsia, and Claviceps (Fig. 1). The phylogenetic analyses clearly indicated that Amphichorda was a monophyletic group, suggesting a common evolutionary origin. The genus Amphichorda had a close genetic relationship with Hapsidospora and Ovicillium, but they were clearly distinguished from their allied genera by forming three separate clades within the Bionectriaceae family. In the phylogenetic tree, the genera Proxiovicillium and Bulbithecium were also closely related to Amphichorda.

In this study, 13 fungal isolates were examined, including four CBS strains. The results showed that these isolates represented nine known species and two new species. The phylogenetic positions of the nine known species were evaluated according to phylogenetic inferences based on the six loci, including Amphichorda coprophila, A. excrementa, A. felina, A. kunmingensis, Clonostachys kunmingensis, C. rosea, C. solani, Sesquicillium buxi, and S. candelabrum (see Table 1, Fig. 1). The phylogenetic analyses also resolved most Amphichorda and Ovicillium lineages in separate terminal branches. It was proposed that two strains, GMBC 3005 and GMBC 3006, which formed a distinct lineage and had a close relationship with A. felina and A. yunnanensis, might be a new species in the genus Amphichorda, named A. guizhouensis. Our analyses further revealed that the newly discovered species, O. pseudoattenuatum (GMBC 3007 and GMBC 3008), were phylogenetically clustered with O. attenuatum and O. sinense, but it was clearly distinguished from the latter two species by forming a well-supported clade in the genus Ovicillium (BSIQ/BSRAx/PP = 82%/-/0.93; Fig. 1).

Taxonomy

Amphichorda guizhouensis Y. Wang & D.X. Tang, sp. nov.

MycoBank No: 857725
Fig. 2

Etymology.

Named after the location Guizhou Province where the species was collected.

Type.

China • Guizhou Province, Anshun city, Xixiu District, Liuguan Village (26.25°N, 106.22°E, 1273 m above sea level), on bird feces, 12 July 2023, Yao Wang (holotype, GMB 3005); ex-type culture, GMBC 3005.

Description.

Sexual morph: Undetermined. Asexual morph: Synnemata arising from bird feces, 1.6–2.0 mm long. Colonies on PDA attaining a diameter of 40–42 mm after a month at 25 °C, white to pinkish, flat, margin entire, reverse yellowish. Hyphae branched, smooth-walled, septate, hyaline, 0.8–2.2 μm wide. Conidiophores arising laterally from hyphae, cylindrical, straight or slightly curved, occasionally branched, hyaline. Conidiogenous cells arising laterally from aerial hyphae, basal portion cylindrical or flask-shaped, erect or irregularly curved, tapering abruptly towards the apex, 6.0–20.8 × 1.8–3.7 (X̄ = 15.2 × 2.6, n = 30) μm. Conidia 2.6–4.0 × 1.8–2.6 (X̄ = 3.1 × 2.2, n = 50) μm, one-celled, smooth-walled, hyaline, subglobose to ellipsoidal, single, often remaining attached to the apex of conidiogenous cells. Chlamydospores not observed.

Figure 2. 

Morphology of Amphichorda guizhouensis A A. guizhouensis on bird feces B Synnemata C, D Colony obverse and reverse on PDA medium E–H Conidiophores, conidiogenous cells and conidia. Scale bars: 5 mm (A); 400 μm (B); 20 mm (C–D); 10 μm (E, G, H); 5 μm (F).

Other material examined.

China • Guizhou Province, Anshun City, Xixiu District, Liuguan Village (26.25°N, 106.22°E, 1269 m above sea level), on bird feces, 12 July 2023, Yao Wang (paratype: GMB 3006); ex-paratype culture, GMBC 3006).

Substrate.

Animal feces.

Distribution.

At present, known only in Anshun City, Guizhou Province, China.

Notes.

Phylogenetic analyses placed A. guizhouensis within the Amphichorda clade, forming a sister lineage to A. felina and A. yunnanensis with strong statistical support (BS/BS/PP = 97%/100%/1; Fig. 1). The species formed a distinct monophyletic group comprising two sampled strains, demonstrating significant genetic divergence from A. felina and A. yunnanensis. Morphologically, our observations unearthed distinct disparities among the three species. Specifically, A. felina exhibited phialides that were consistently flask-shaped, while A. guizhouensis featured phialides that were either cylindrical or flask-shaped. In contrast, A. yunnanensis possessed phialides ranging from ampulliform to flask-shaped. A particularly notable characteristic of A. guizhouensis was its relatively elongated phialides (6.0–20.8 × 1.8–3.7 µm). This unique morphological trait served as a crucial diagnostic feature, enabling clear differentiation of A. guizhouensis from other species within the Amphichorda genus (see Table 2).

Table 2.
Download as
CSV
XLSX

Hosts/substrates and asexual morphology of Amphichorda.

Species Host/Substrate Conidiophores Phialides (μm) Conidia (μm) References
Amphichorda cavernicola Bird feces; soil; plant debris; animal feces; bat guano Cylindrical, straight or slightly curved, occasionally branched Fusiform or ellipsoidal, straight or irregularly bent, 4.5–8.0 × 2.0–3.0 Broadly ellipsoidal to subglobose, 2.5–4.0 × 2.0–3.5 Zhang et al. 2020
Amphichorda coprophila Chipmunk, rabbit and porcupine dung Straight or flexuous, unbranched, bearing lateral or terminal conidiogenous cells, arranged singly or in whorls Flask-shaped, usually with a strongly bent neck, 6–10 × 2–2.5 Subglobose to somewhat ellipsoidal, 3.5–5.5 × 2–3 Guerra-Mateo et al. 2023
Amphichorda excrementa Animal feces Cylindrical, straight or slightly curved, occasionally branched Occasionally solitary, mostly in whorls of 2–3, basal portion cylindrical or flask-shaped, usually curved, 4.1–13.9 × 1.3–2.1 Globose to elliptical 1.7–3.0 × 1.2–2.5 Wang et al. 2024
Amphichorda felina Pupae of Anaitis efformata; rabbit dung; moudy leaves; porcupine dung; cat dung Straight Solitarily or in small groups, consisting of a swollen, flask-shaped or curved, occasionally elongate basal part, 1.5–8.5 × 1.8–2.9 Subglobose, ellipsoidal or ovoidal, sometimes with a pointed base, 2.5–4.7 × 2–3.5 Wang et al. 2024
Amphichorda guana Bat guano Straight or slightly curved Fusiform or ellipsoidal, straight or irregularly bent, 7–10 × 2–3 Broadly ellipsoid to subglobose, 4.5–5.5 × 3.5–5 Zhang et al. 2017
Amphichorda guizhouensis Animal feces Cylindrical, straight or slightly curved, occasionally branched Basal portion cylindrical or flask-shaped, erect or irregularly curved, tapering abruptly towards the apex, 6.0–20.8 × 1.8–3.7 Subglobose to ellipsoidal, 2.6–4.0 × 1.8–2.6 In this study
Amphichorda kunmingensis Animal feces - Solitary, occasionally in simple whorls, basal portion cylindrical or fusiform, straight or irregularly bent, 6.1–17.5 × 1.4–2.9 Globose to elliptical 2.3–4.2 × 1.6–3.0 Wang et al. 2024
Amphichorda littoralis Sediments; fragment of floating rubber tire Straight or flexuous, commonly unbranched, bearing lateral or terminal conidiogenous cells, arranged singly or in whorls of 2–4 Flask-shaped, usually with a strongly bent neck, 6–10 (–11.5) × 1.5–2 Subglobose, 3–4 × 2.5–3 Guerra-Mateo et al. 2023
Amphichorda monjolensis On PDA plate consumed by an insect Cylindrical, bearing one or more conidiogenous cells, straight or slightly bent, solitary or synnematous, sometimes branched Flask-shaped, straight or irregularly bent, 3.1–6.1 × 2.7–5.1 Holoblastic, 2.8–3.7 × 1.8–2.9 Leão et al. 2024
Amphichorda yunnanensis Wing surfaces
of Rhinolophus 		Cylindrical, straight or slightly curved, branched Monoblastic to polyblastic, ampulliform to flask-shaped, 4–12 × 1–4 Globose to oval, slightly ellipsoid, 2–5 × 2–4 Liu et al. 2023

Ovicillium pseudoattenuatum Y. Wang & D.X. Tang, sp. nov.

MycoBank No: 857726
Fig. 3

Etymology:

“Pseudoattenuatum” refers to morphologically resembling Ovicillium attenuatum, but phylogenetically distinct.

Type.

Laos • Vientiane City, Mekong Riverside Park (17.96°N, 102.60°E, 674 m above sea level), from soil on the forest floor, 11 August 2024, Yao Wang (holotype as dried culture GMB 3007); ex-type culture GMBC 3007.

Description.

Sexual morph: Undetermined. Asexual morph: Colonies on PDA reaching 23–25 mm in diameter in 7 days at 25 °C, white to pinkish; reverse yellowish. Hyphae branched, smooth-walled, septate, hyaline, 1.2–2.8 μm wide. Conidiophores hyaline, smooth-walled, with single phialide or whorls of 2–5 phialides or verticillium-like directly from hyphae, up to 500 μm long. Phialides terminal or lateral, straight, somewhat inflated base, attenuated from the middle, sometimes undulated near the tip, 16.0–37.5 × 1.5–2.4 (X̄ = 26.8 × 2.0, n = 50) μm. Conidia smooth-walled, hyaline, ellipsoidal to cylindrical, 3.2–4.0 × 1.7–3.2 (X̄ = 3.7 × 2.3, n = 50) μm, aggregated in large globose to subglobose heads. Crystals absent. Chlamydospores absent.

Figure 3. 

Morphology of Ovicillium pseudoattenuatum A, B Colony obverse and reverse on PDA medium C–I Conidiophores, phialides and conidia J Conidia. Scale bars: 10 mm (A–B); 100 μm (C); 50 μm (D–E); 25 μm (F–I); 10 μm (J).

Other material examined.

Laos • Oudomxay Province, Muang Xay District, Nam Kat Yorla Pa Resort (20.71°N, 102.11°E, 708 m above sea level), from soil on the forest floor, 14 August 2024, Yao Wang (living culture GMBC 3008).

Substrate.

Soil.

Distribution.

Laos.

Notes.

Ovicillium pseudoattenuatum, isolated from forest floor soil, forms a distinct phylogenetic lineage within the Ovicillium genus. Multilocus phylogenetic analyses reveal its close relationship with O. attenuatum and O. sinense, supported by strong statistical values (BSIQ/BSRAx/PP = 82%/79%/0.93). Morphologically, while sharing the characteristic undulated phialide tips with O. attenuatum, O. pseudoattenuatum differs significantly in microscopic dimensions: it possesses smaller phialides (16.0–37.5 × 1.5–2.4 μm vs 25–50 × 1.7–3.3 μm) and more compact conidia (3.2–4.0 × 1.7–3.2 μm vs 3.5–5 × 2.5–3.8 μm). Distinct from O. sinense, which exhibits even smaller reproductive structures (phialides 16.2–25.8 × 1.7–2.4 μm; conidia 2.1–2.9 × 1.1–1.7 μm), O. pseudoattenuatum is further characterized by its unique ellipsoidal to cylindrical conidial morphology, a diagnostic feature distinguishing it from all known Ovicillium species (Table 3).

Table 3.
Download as
CSV
XLSX

Hosts/substrates and asexual morphology of Ovicillium.

Species Host/Substrate Conidiophores (μm) Phialides (μm) Conidia (μm) Crystals Chlamydospores (μm) References
Ovicillium asperulatum (Synonym: O. napiforme) Soil; wood of Sorbus aria Simple or mostly branched, bearing whorls of 2–4 phialides, up to 105 long, with cell walls usually thicker than those of the vegetative hyphae Straight or slightly bent, acicular, 28–68 long, 1–2 wide at the base, with minute collarette and distinct periclinal thickening at the apex Globose, 3–4(–5) diameter, chromophilic Absent Abundant, subglobose or oval, 5–10 × 5–9 Giraldo et al. 2012; Zare and Gams 2016
Ovicillium attenuatum Auricularia sp. Erect, 150–500 tall, with cyanophilic encrustation, usually with whorls, but sometimes also solitary phialides Aculeate, attenuated from the middle, mostly undulated near the tip, rather cyanophilic, measuring 25–50 × 1.7–3.3 Oval to subglobose, strongly cyanophilic, measuring 3.5–5 × 2.5–3.8, aggregated in large globose to subglobose heads Absent Absent Zare and Gams 2016
Ovicillium oosporum Theobroma gileri Erect, solitary and verticillate, with slightly pigmented base producing solitary or verticillate phialides of up to five per node Verticillate, measuring 20–50 × 1.2–2.2 Subglobose, oval to broadly oval, in some strains with a basal protrusion, cyanophilic, measuring 4–6 × 2.5–4 Absent Present or absent Zare and Gams 2016
Ovicillium pseudoattenuatum Soil Single phialide or whorls of 2–5 phialides or verticillium-like directly from hyphae, up to 500 long Straight, somewhat inflated base, attenuated from the middle, sometimes undulated near the tip, 16.0–37.5 × 1.5–2.4 Ellipsoidal to cylindrical, 3.2–4.0 × 1.7–3.2, aggregated in large globose to subglobose heads Absent Absent In this study
Ovicillium sinense Pupa (Lepidoptera) Single phialide or whorls of 2–5 phialides or verticillium-like, 17.0–21.7 × 2.3–3.0 Cylindrical, somewhat inflated base, 16.2–25.8 × 1.7–2.4, tapering to a thin neck Globose to ovoid, 2.1–2.9 × 1.1–1.7, aggregated in large globose to subglobose heads – – Chen et al. 2024
Ovicillium subglobosum Soil Erect, solitary and verticillate with up to four phialides per node Measuring 25–55 × 1.5–2.2, producing conidia in large globose heads. Subglobose with an inconspicuous protrusion at the base, rather cyanophilic, measuring 3.5–5.5 × 3.5–4.5 Absent Absent Zare and Gams 2016
Ovicillium variecolor Soil Erect, mostly branched, bearing whorls of 2–5 phialides, up to 290 long, with walls usually thicker than those of the vegetative hyphae Straight, acicular, 18–95 long, 1–2 wide at the base, with periclinal thickening at the apex, collarette inconspicuous; some phialidic conidiogenous cells without a basal septum (adelophialides) Subglobose or ovoid, 3–4(–5) × 2–4, slightly apiculate base, chromophilic, arranged in slimy heads – Absent Giraldo et al. 2012

Discussion

Amphichorda species exhibit exceptional ecological plasticity, colonizing diverse substrates including caves, marine sediments, soil, animal waste, and even biotic surfaces such as the wings of bats (Rhinolophus spp.). Although their primary niche is coprophilous (dung-associated)—evidenced by the recent discovery of A. guizhouensis and most congeners in animal excrement—their adaptability is further exemplified by colonization of specialized microhabitats. For instance, A. felina has been isolated from decomposing leaves and Anaitis efformata pupae (de Hoog 1972; Wang et al. 2024), while A. yunnanensis inhabits bat wing surfaces (Liu et al. 2023). Notably, three Hapsidospora species (H. globosa, H. inversa, and H. stercoraria) also exhibit obligate coprophily. This parallel ecological specialization between Amphichorda and Hapsidospora underscores their phylogenetic affinity, suggesting either a shared ancestral adaptation to fecal substrates or convergent niche evolution within the Bionectriaceae family.

The broad ecological range of Amphichorda is accompanied by subtle morphological distinctions, complicating taxonomic differentiation among cryptic species. Consequently, molecular data play a pivotal role in delineating species boundaries. Recent studies have employed multi-locus sequence data (ITS, nrSSU, nrLSU, tef1α, rpb1, and rpb2) to resolve phylogenetic relationships within the genus (Zhang et al. 2017, 2020; Guerra-Mateo et al. 2023; Liu et al. 2023; Leão et al. 2024; Wang et al. 2024). However, GenBank records (https://www.ncbi.nlm.nih.gov, accessed 20 February 2025) remain incomplete for nrSSU, rpb1, and rpb2 in this group. To address this gap, we generated comprehensive molecular datasets from five distinct Amphichorda species. These data enrich existing genomic resources and enhance the resolution of phylogenetic reconstructions, providing deeper insights into the evolutionary history of the genus.

Our multi-locus phylogeny (nrSSU-ITS-nrLSU-tef1α-rpb1-rpb2) revealed a close genetic relationship among Amphichorda, Hapsidospora, Ovicillium, Proxiovicillium, and Bulbithecium. Consistent with Leão et al. (2024), Amphichorda and Hapsidospora form a well-supported sister clade within Bionectriaceae (BSIQ/BSRAx/PP = 93%/99%/1; Fig. 1). Additionally, Ovicillium clustered with Proxiovicillium + Bulbithecium as a distinct lineage, while maintaining a close genetic affinity with Amphichorda. Although earlier studies positioned Ovicillium as sister to Proxiovicillium (Hou et al. 2023; Leão et al. 2024), our expanded sampling of seven Ovicillium species [compared to two in Hou et al. (2023) and three in Leão et al. (2024)] yielded more stable topological relationships among these genera. Morphological traits further corroborate these genetic linkages: Ovicillium species produce large globose conidial heads, Proxiovicillium forms conidia in elongated chains, and Bulbithecium exhibits slimy conidial heads arranged in chains.

The ongoing discovery of new species in biodiversity studies is essential to our comprehension of the complexity of ecosystems and the development of life. The discovery of novel species remains pivotal for elucidating ecosystem complexity and evolutionary trajectories. Here, we propose two new species—A. guizhouensis (isolated from animal feces) and O. pseudoattenuatum (from soil)—based on integrated morphological and phylogenetic evidence. A comparative analysis of morphological traits across all Amphichorda and Ovicillium members was conducted to refine taxonomic boundaries. This study not only clarifies evolutionary relationships within the two genera but also advances the systematic understanding of biodiversity in Bionectriaceae.

Acknowledgments

In this section, you can acknowledge any support given which is not covered by the author contribution or funding sections. This may include administrative and technical support, or donations in kind (e.g., materials used for experiments).

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This research was jointly supported by the National Natural Science Foundation of China under grants [32460004] and [32200013], the High-level Innovation Talents (No. GCC[2023]048), and the Guizhou Provincial Scientific and Technologic Innovation Base (No. [2023]003).

Author contributions

All authors have contributed equally.

Author ORCIDs

Yao Wang https://orcid.org/0000-0002-1262-6700

Hui Chen https://orcid.org/0009-0008-0291-3571

AuthorName https://orcid.org/0000-0002-4333-9106

Data availability

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

References

  • Carmichael JW, Kendrick WB, Conners IL, Sigler L (Eds) (1980) Genera of Hyphomycetes. University of Alberta Press, 396 pp.
  • Chen MJ, Huang B, Li ZZ, Spatafora JW (2013) Morphological and genetic characterisation of Beauveria sinensis sp. nov. from China. Mycotaxon 124(1): 301–308. https://doi.org/10.5248/124.301
  • Darriba D, Taboada GL, Doallo R, Posada DJ (2012) ModelTest 2: More Models, New Heuristics and Parallel Computing. Nature Methods 9(8): 772. https://doi.org/10.1038/nmeth.2109
  • de Hoog GS (1972) The genera Beauveria, Isaria, Tritirachium and Acrodontium gen. nov. Studies in Mycology 1: 1–41.
  • Fries EM (1825) Systema Orbis Vegetabilis: Primas Lineas Novae Constructionis Periclitatur; E Typographia Academica. Lunde Volume 1.
  • Fries EM (1832) Systema Mycologicum: Sistens Fungorum Ordines, Genera et Species, Huc Usque Cognitas, Quas ad Normam Methodi Naturalis Determinavit; Sumtibus Ernesti Mauritii: Gryphiswaldia. Germany Volume 3.
  • Giraldo A, Gene J, Cano J, de Hoog S, Guarro J (2012) Two new species of Acremonium from Spanish soils. Mycologia 104(6): 1456–1465. https://doi.org/10.3852/11-402
  • Guerra-Mateo D, Gené J, Baulin V, Cano-Lira JF (2023) Phylogeny and taxonomy of the genus Amphichorda (Bionectriaceae): An update on Beauveria-like strains and description of a novel species from marine sediments. Diversity 15(7): 795. https://doi.org/10.3390/d15070795
  • Hoang DT, Chernomor O, von Haeseler A, Minh BQ, Vinh LS (2017) UFBoot2: Improving the ultrafast bootstrap approximation. Molecular Biology and Evolution 35(2): 518–522. https://doi.org/10.1093/molbev/msx281
  • Hopple JS (1994) Phylogenetic investigations in the genus Coprinus based on morphological and molecular characters. Ph.D. Dissertation, Duke University, Durham, NC, USA.
  • Hou LW, Giraldo A, Groenewald JZ, Rämä T, Summerbell RC, Huang GZ, Cai L, Crous PW (2023) Redisposition of acremonium-like fungi in Hypocreales. Studies in Mycology 105(1): 23–203. https://doi.org/10.3114/sim.2023.105.02
  • Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A, Jermiin LS (2017) ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589. https://doi.org/10.1038/nmeth.4285
  • Leão AF, Condé TO, Dutra YLG, Rosado AWC, Grazziotti PH, Neves SC, Fraga LMS, Kasuya MCM, Pereira OL (2024) Amphichorda monjolensis sp. nov., a new fungal species isolated from a Brazilian limestone cave, with an update on acremonium-like species in Bionectriaceae. Brazilian Journal of Microbiology 55(2): 1569–1585. https://doi.org/10.1007/s42770-024-01289-y
  • Lechat C, Fournier J (2019) New insights into Stilbocrea (Hypocreales, Bionectriaceae): Recognition of S. colubrensis, a new species from Martinique (French West Indies), and observations on lifestyle and synnematous asexual morphs of S. gracilipes and S. macrostoma. Ascomycete. Org : Revue Internationale pour la Taxinomie des Ascomycota 11(6): 183–190. https://doi.org/10.25664/ART-0272
  • Liu ZY, Liang ZQ, Whalley AJS, Yao YJ, Liu AY (2001) Cordyceps brittlebankisoides, a new pathogen of grubs and its anamorph, Metarhizium anisopliae var. majus. Journal of Invertebrate Pathology 78(3): 178–182. https://doi.org/10.1006/jipa.2001.5039
  • Liu XF, Tibpromma S, Hughes AC, Chethana KWT, Wijayawardene NN, Dai DQ, Du TY, Elgorban AM, Stephenson SL, Suwannarach N, Xu JC, Lu L, Xu RF, Maharachchikumbura SSN, Zhao CL, Bhat DJ, Sun YM, Karunarathna SC, Mortimer PE (2023) Culturable mycota on bats in central and southern Yunnan Province, China. Mycosphere : Journal of Fungal Biology 14(1): 497–662. https://doi.org/10.5943/mycosphere/14/1/7
  • Minh BQ, Schmidt HA, Chernomor O, Schrempf D, Woodhams MD, von Haeseler A, Lanfear R (2020) IQ-TREE 2: New models and efficient methods for phylogenetic inference in the genomic era. Molecular Biology and Evolution 37(5): 1530–1534. https://doi.org/10.1093/molbev/msaa015
  • Rehner SA, Buckley EA (2005) Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: Evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97(1): 84–98. https://doi.org/10.3852/mycologia.97.1.84
  • Rehner SA, Minnis AM, Sung GH, Luangsa-ard JJ, Devotto L, Humber RA (2011) Phylogeny and systematics of the anamorphic, entomopathogenic genus Beauveria. Mycologia 103(5): 1055–1073. https://doi.org/10.3852/10-302
  • Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61(3): 539–542. https://doi.org/10.1093/sysbio/sys029
  • Spatafora JW, Sung GH, Sung JM, Hywel‐Jones NL, White Jr JF (2007) Phylogenetic evidence for an animal pathogen origin of ergot and the grass endophytes. Molecular Ecology 16(8): 1701–1711. https://doi.org/10.1111/j.1365-294X.2007.03225.x
  • Sung GH, Hywel-Jones NL, Sung JM, Luangsa-ard JJ, Shrestha B, Spatafora JW (2007) Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Studies in Mycology 57: 5–59. https://doi.org/10.3114/sim.2007.57.01
  • 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
  • Wang ZQ, Zhao J, Dong QY, Wang Y, Lu YL, Luo R, Yu H (2024) Multi-locus molecular phylogenetic analysis reveals two new species of Amphichorda (Bionectriaceae, Hypocreales). MycoKeys 106: 287–301. https://doi.org/10.3897/mycokeys.106.117205
  • White TJ, Bruns TD, Lee SB, Taylor JW (1990) Amplifcation and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (Еds) PCR protocols: a guide to methods and applications. Academic Пресс, 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
  • Zhang ZF, Zhou SY, Eurwilaichitr L, Ingsriswang S, Raza M, Chen Q, Zhao P, Liu F, Cai L (2020) Culturable mycobiota from Karst caves in China II, with descriptions of 33 new species. Fungal Diversity 106(1): 29–136. https://doi.org/10.1007/s13225-020-00453-7
login to comment