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Research Article
Two new species of Nectriaceae (Hypocreales, Sordariomycetes) from Yunnan, China
expand article infoHua Zheng‡§, Xinwen Dai§, Haiyan Li‡, Zefen Yu§
‡ Kunming University of Science and Technology, Kunming, China
§ Yunnan University, Kunming, China

Corresponding author: Haiyan Li ( lhyxrn@163.com )

Corresponding author: Zefen Yu ( zfyu2021@163.com )

Academic editor: Danushka Sandaruwan Tennakoon
© 2024 Hua Zheng, Xinwen Dai, Haiyan Li, Zefen Yu.
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: Zheng H, Dai X, Li H, Yu Z (2024) Two new species of Nectriaceae (Hypocreales, Sordariomycetes) from Yunnan, China. MycoKeys 108: 269-285. https://doi.org/10.3897/mycokeys.108.130098
Open Access

Abstract

Nectriaceae is a highly diverse family, and members have a worldwide distribution, particularly in warm temperate to tropical regions. During the survey of fungal diversity in different habitats in Yunnan province, China, two new species isolated from soil and air respectively, namely Atractium yunnanense sp. nov. and Nalanthamala xishuangbannaensis sp. nov., were proposed based on morphological comparisons and the multi-gene phylogenetic analyses of combined ITS, LSU, rpb2, and tub2 sequence data. Phylogenetically, both species clustered in a monophyletic clade within Nectriaceae with strong support. A. yunnanense is characterized by synnematous conidiophores, pale olivaceous-green, clavate to oblong-ellipsoidal, multi-septate conidia, and pale olivaceous-green chlamydospores. N. xishuangbannaensis has acremonium-like or penicillium-like conidiophores and either obovate or ellipsoidal, cylindrical or fusiform conidia. Full descriptions, illustrations, and a phylogenetic tree showing the phylogenetic position of the two new species were provided.

Key words

Air-borne or soil-borne fungi, fungal diversity, Hypocreales, phylogeny, taxonomy

Introduction

Nectriaceae was originally established based on Nectria (Fr.) Fr. and assigned in Hypocreales (Tulasne and Tulasne 1865). Thereafter, the taxonomy of Nectriaceae has undergone several revisions (Dennis 1960; Rossman et al. 1999; Lombard et al. 2015). This family is characterized by uniloculate and verrucose ascomata by reaction in KOH and lactic acid, and unitunicate, 2–8-spored asci with globose, ellipsoid to long-fusiform ascospores. Additionally, it is associated with phialidic asexual morphs producing amerosporous to phragmosporous conidia (Lombard et al. 2015). In a recent study, Perera et al. (2023) proceeded to update DNA sequence-based phylogeny for the order Hypocreales, and accepted 77 genera into Nectriaceae, including Atractium Link and Nalanthamala Subram. Nectriaceae is a highly diverse group, and members have a cosmopolitan distribution, particularly in the warm temperate and tropical regions (Rossman et al. 1999; Chaverri et al. 2011; Lombard et al. 2015). Particularly, this family has ecological and economic significance. The majority of the species are soil-borne saprobes or weak to virulent plant pathogens, and include important plant pathogens of economically important plants worldwide, such as Ilyonectria species and Fusarium species (Jayawardena et al. 2019a, 2019b). Moreover, several species have also been reported as important opportunistic pathogens of humans (Chang et al. 2006; Guarro 2013; Köhler et al. 2015, 2017; Hoenigl et al. 2021). For instance, a patient with acute myelogenous leukemia experienced a disseminated cerebellar Fusarium infection, which magnetic resonance imaging and a CT scan revealed to be a left cerebellar focus of Fusarium solani (Vincent et al. 2003).

Atractium was introduced by Link (1809) with A. stilbaster Link as the type species. So far, there are 28 epithets for Atractium as listed in the Index Fungorum (https://www.indexfungorum.org; retrieval in June 2024). Of them, five species were accepted in the genus, while the placement of the other 23 species remains uncertain due to disagreement with the emended generic diagnosis or an inability to locate authentic material (Gräfenhan et al. 2011; Bao et al. 2023). Gräfenhan et al. (2011) conducted a comprehensive phylogenetic reassessment of nectriaceous fungi, and accepted three asexual species A. crassum (Wollenw.) Seifert & Gräfenhan, A. holubovae (Seifert, S.J. Stanley & K.D. Hyde) Seifert and A. stilbaster in Atractium. In a recent study, a sexual morph species Varicosporella aquatica Lechat & J. Fourn. was transferred to Atractium, namely A. aquatica (Lechat & J. Fourn) D.F. Bao, K.D. Hyde & Z.L. Luo, and a new species A. fusiformis D.F. Bao, K.D. Hyde & Z.L. Luo was introduced based on morphological and phylogenetic analysis (Bao et al. 2023). The asexual morph of Atractium is characterized by synnematous conidiophores, monophialidic, subulate conidiogenous cells with septate, clavate, obovoid or gently curved conidia, and its sexual morphs fit well with the generic concepts of Nectriaceae (Bao et al. 2023). Currently, single Atractium species (A. fusiformis) was reported as saprobe from China among the five accepted species.

Nalanthamala was proposed for N. madreeya Subram. and characterized by mononematous or aggregated conidiophores, singly or in whorls produced phialides, and elliptical to oval or lenticular conidia arranged in chains (Subramanian 1956). Gams (1975) placed Fusidium squamicola Berk. & Broome in Nalanthamala based on morphological resemblance of conidia, namely N. squamicola (Berk. & Broome) W. Gams (Gams 1975). Schroers et al. (2005) demonstrated that Nalanthamala belongs in Nectriaceae using LSU rDNA sequences, and accepted three species, including N. diospyri (Crandall) Schroers & M.J. Wingf., N. psidii (Sawada & Kurosawa) Schroers & M.J. Wingf., and N. vermoesenii (Biourge) Schroers. Subsequently, Rossman et al. (2013) proposed Nalanthamala over Rubrinectria Rossman & Samuels, and included N. olivacea (Seaver) Rossman in this genus. However, Crous et al. (2021) transferred N. squamicola to the newly established genus Caespitomonium Crous in Bionectriaceae, named after C. squamicola (Berk. & Broome) Crous. Currently, there are five species in this genus Nalanthamala (Wijayawardene et al. 2020; Crous et al. 2021). Interestingly, the known Nalanthamala species are also associated with wilt and blight diseases of several economically important crops (Schroers et al. 2005; Rossman et al. 2013). However, a single species, N. psidii, has been recorded from China (Schroers et al. 2005).

Southwest China is one of the high biodiversity hotspots in the word (Myers et al. 2000; Orme et al. 2005; Mi et al. 2021). Particularly, there is a high fungal diversity in Yunnan province (Feng and Yang 2018; Zheng et al. 2021b). In the last decade or so, our team discovered many new fungal species during the investigation of fungal diversity in different habitats in Yunnan (Zheng et al. 2019, 2020, 2021a, 2022; Qiao et al. 2020, 2021; Yu et al. 2022), which increases knowledge of this important ecological area. In this study, two new taxa in Nectriaceae were discovered from Yunnan, and described as new species of Atractium and Nalanthamala, namely A. yunnanense sp. nov. and N. xishuangbannaensis sp. nov., based on morphological characteristics and multi-gene phylogenetic analysis.

Materials and methods

Sample collection, fungal isolation and morphological characterization

Soil samples were collected from Huize county, Yunnan province. Samples were preserved in sterile plastic bags, labeled and transported to the laboratory at 4 °C. The dilution coating method was used to isolate fungal strains from soils as described by Lv et al. (2022). The fungi in air located at Xishuangbanna, Yunnan province, were collected by means of the MAS-100 ISO MH Microbial Air Sampler (Merck Millipore, Germany). Ten liters of air flowed through the surface of each Rose Bengal agar (Guangdong Huankai Microbial Sci and Tech, China) plate which placed in the air sampling equipment. After incubation at 25 °C for 5 days, representative colonies were picked up with a sterilized needle and maintained on potato dextrose agar (PDA; 200 g potato, 20 g dextrose, 18 g agar, 1000 ml distilled water) plates. The pure strains were incubated on PDA and cornmeal agar (CMA; 20 g cornmeal, 18 g agar, 1000 ml distilled water) for observing morphological characteristics of colonies. Microscopic characteristics growing on CMA were examined and captured by an Olympus BX51 microscope connected to a DP controller digital camera, and sterile water was used as a mounting medium for microscopy.

The pure cultures were deposited in the Herbarium of the Laboratory for Conservation and Utilization of Bio-Resources, Yunnan University, Kunming, Yunnan, P.R. China (YMF), China General Microbiological Culture Collection Center (CGMCC), the Guangdong Microbial Culture Collection Center (GDMCC) and Japan Collection of Microorganisms (JCM). MycoBank numbers were obtained in MycoBank database (https://www.mycobank.org/) for the newly-described taxa.

DNA extraction, PCR amplification and sequencing

Total DNA was extracted from fresh mycelia grown on PDA for 7 days, as described by Turner et al. (1997). The primer pairs ITS5/ITS4 (White et al. 1990), LR0R/LR7 (Vilgalys and Hester 1990), rpb2-5F2/rpb2-7CR (Liu et al. 1999), and TUB-2Fd/TUB4RD (Woudenberg et al. 2009) were used for amplification of the internal transcribed spacers (ITS), the large subunit nuclear ribosomal RNA gene (LSU), the RNA polymerase II second largest subunit gene (rpb2), and the beta-tubulin gene (tub2), respectively. The PCR amplifications were performed in 25 µl reaction volumes containing 1.0 µl DNA template, 1.0 µl of each forward and reverse primers, 12.5 µl 2 × Master Mix, and 9.5 µl ddH2O. The PCR products were confirmed on 1% agarose electrophoresis gels, and the positive products were sent to Tsingke Biotechnology Company (Kunming, China) for purification and sequencing. Newly obtained sequences were deposited in the GenBank database at the National Center for Biotechnology Information (NCBI) and the accession numbers are listed in Table 1.

Table 1.
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Species, strains and their corresponding GenBank accession numbers of sequences used for phylogenetic analyses.

Species Strains GenBank accession no.
ITS LSU rpb2 tub2
Atractium aquatica CBS126103T KP192669 KP192671 – –
CBS138883 KP192668 KP192670 – –
Atractium crassum CBS 180.31T KM231790 U88110 – KM232049
Atractium fusiformis KUNCC22-12521 OP876729 OP875082 – OQ025196
KUNCC22-12523T OP876725 OP875078 – OQ025192
KUNCC22-12452 OP876727 OP875080 – OQ025195
Atractium stilbaster CBS 410.67T KM231791 KM231654 – KM232050
CBS 783.85 KM231792 KM231655 – KM232051
Atractium yunnanense YMF 1.06524T OM985710 PP915817 PP928787 PP928793
H77 PP915812 PP915818 PP928788 PP928794
H102 PP915813 PP915819 PP928789 PP928795
Cosmospora aquatica MFLUCC13-0884T NR_168211 MK828238 MN194021 –
Cosmospora butyri DAOM216335 JN942831 JN938895 –
Cosmospora cymosa CBS 762.69T NR_111605 NG_058891 HQ897778 –
Fusarium coffeatum CBS 635.76 MH861016 AY213706 KU604328 –
Fusarium equiseti NL19-25004 MZ890491 MZ890346 MZ921701 –
Fusarium incarnatum CBS 161.25 MH854830 MH866331 MN170381 –
Fusarium proliferatum CBS 263.54 KM231815 KM231684 KM232383 –
CBS 153.27 MH854910 MH866404 – –
Ilyonectria capensis CBS 132815T NR_152887 MH878251 KM232336 –
Ilyonectria coprosmae CBS119606 JF735260 KM515910 KM232338 –
Nalanthamala diospyri CBS 430.89 AY554209 AY554248 – AY554228
Nalanthamala eleanorwilliamsiae BRIP 66236a OQ917077 – – –
Nalanthamala olivacea CBS 102268 AY554219 AY554244 – AY554238
Nalanthamala psidii CBS 687.97 AY554208 – – AY554227
CBS 110184 AY554207 – – AY554226
CBS 110188 AY554206 – – AY554225
Nalanthamala vermoesenii CBS 110893T AY554214 AY554246 – AY554233
CBS 137.24 AY554217 AY554260 – AY554236
Nalanthamala xishuangbannaensis YMF 1.05062T PP915809 PP915814 PP928784 PP928790
B413 PP915810 PP915815 PP928785 PP928791
B425 PP915811 PP915816 PP928786 PP928792
Nectria balansae CBS 124070 JF832652 JF832710 – JF832907
AR4635 JN995622 JN939838 – JF832908
Neonalanthamala graminearum CGMCC3.25240T OQ733285 – OQ716735 OQ716739
S2 OQ733286 – OQ716736 OQ716740
S4 OQ733287 – OQ716737 OQ716741
Neonectria aquatica KUNCC22-12462T OP876733 OP875087 – OQ025197
Neonectria lugdunensis CBS 250.58 KM515893 KM515938 – –
Neonectria ramulariae CBS 151.29 JF735313 AY677333 DQ789792 –
CBS 182.36 JF735314 HM042435 DQ789793 JF735439
Sarocladium summerbellii CBS 430.70 MH859780 MH871543 – –

Notes: T = Ex-type; New isolates are in bold; The line “–” represents the absence of GenBank record.

Sequence alignment and phylogenetic analysis

Initially, the newly-generated sequences were subjected to the GenBank nucleotide database (https://blast.ncbi.nlm.nih.gov/) using BLAST searches to determine closely related taxa, including Atractium species and Nalanthamala species. To further determine the phylogenetic position of these strains, a multi-locus phylogenetic analysis was performed based on ITS, LSU, rpb2 and tub2. Alignments of different loci, including the sequences obtained from this study and sequences downloaded from GenBank, were initially aligned with ClustalX v1.83 (Thompson et al. 1997). The resulting alignments were subsequently checked and refined using BioEdit v7.0.4.1 (Hall 1999). The concatenation of the four loci was processed with BioEdit, and converted to a NEXUS file using MEGA v10 (Kumar et al. 2018). The concatenated sequence matrix contained 3,064 nucleotide positions from the four loci (633 from ITS, 886 from LSU, 932 from rpb2, and 613 from tub2), and was uploaded to TreeBASE (https://www.treebase.org; submission number: S31491).

Maximum Likelihood (ML) and the Bayesian Inference (BI) methods were used for the phylogenetic construction in this study. The ML analysis was performed by RAxML v8.0.9 (Stamatakis 2006) using the GTR-GAMMA model with rapid bootstrap analysis, followed by 1,000 ultrafast bootstrap replicates to estimate bootstrap support values (BS). The BI analysis was conducted with MrBayes v3.2.2 (Ronquist et al. 2012) with NEXUS file. The jModelTest v2.0 (Posada 2008) was used to carry out statistical selection of best-fit models of nucleotide substitution using the corrected Akaike information criterion (AIC), and the estimated best-fit model was GTR+F+I+G4. Markov Chain Monte Carlo (MCMC) simulations were used for 1,000,000 generations with a sampling frequency of every 500th generations. The initial 25% of sample trees were treated as burn-in and discarded. The remaining trees were used to calculate the Bayesian posterior probabilities (BP). The phylogenetic trees were visualized using FigTree v1.4.3 and subsequently edited in Adobe Photoshop.

Results

Phylogenetic analysis

The concatenated dataset comprised 41 taxa (including our strains) representing eight genera in Nectriaceae (Hypocreales), with Sarocladium summerbellii (CBS 430.70) as the outgroup. The phylogenetic trees using ML and BI analyses were consistent and strongly supported in most branches. The topology of the phylogenetic tree is shown in Fig. 1, with maximum likelihood bootstrap support values (BS) ≥ 75% and Bayesian posterior probabilities (BP) ≥ 0.90 were shown at the nodes. In this tree, three isolates (YMF 1.06524, H77 and H102) clustered in a well-separated clade with a high support value (BP/BS = 1.00/100) and sister to Atractium crassum (CBS 180.31) with strong support (BP/BS = 1.00/100). The other three isolates (YMF 1.05062, B413 and B425) formed a distinct clade with a high support value (BP/BS = 1.00/100), and the clade grouped together with Nalanthamala species, including N. vermoesenii, N. olivacea and N. psidii. Therefore, two new taxa, Atractium yunnanense sp. nov. and Nalanthamala xishuangbannaensis sp. nov., are proposed according to the phylogenetic analysis.

Figure 1. 

Phylogenetic tree inferred from a Maximum likelihood analysis based on a concatenated alignment of ITS, LSU, rpb2 and tub2 sequences of some representing species in Nectriaceae. The Bayesian posterior probabilities (BP) ≥ 0.9 and RAxML bootstrap support values (BS) ≥ 75% were shown at the nodes (BP/BS). Strains obtained in this study are shown in blue font. Ex-type strains are marked by a T after the strain number. The tree was rooted to Sarocladium summerbellii (CBS 430.70).

Taxonomy

Sordariomycetes O.E. Erikss. & Winka

Hypocreales Lindau

Nectriaceae Tul. & C. Tul.

Atractium Link, Mag. Gesell. naturf. Freunde, Berlin 3(1–2): 10 (1809)

≡ Varicosporella Lechat & J. Fourn.

Atractium yunnanense H. Zheng & Z.F. Yu, sp. nov.

MycoBank No: MB854385
Fig. 2

Etymology

Refers to the Yunnan province where the holotype was collected.

Figure 2. 

Atractium yunnanense (YMF 1.06524, holotype) A colony on PDA after 14 days of inoculation at 28 °C B colony on CMA after 3 days of inoculation at 28 °C C conidiophores D conidia E chlamydospores. Scale bars: 1.9 cm (A, B); 10 µm (C–E).

Type

China • Yunnan province, Huize county; isolated from soil in karst rocky desertification area; Oct 2020; Z.F.Yu, preserved by lyophilization (a metabolically inactive state) in State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan (YMF 1.06524, holotype); ex-type living culture: CGMCC 3.20977, other living cultures: GDMCC 3.734; JCM 39337.

Description

Sexual morph not observed. Asexual morph on CMA. Hyphae 1.5–3 μm wide, pale olivaceous-green, occasionally branched, septate, smooth-walled. Conidiophores sometimes aggregated into synnemata, nonstromatic, macronematous, mononematous, cylindrical or subulate, straight or flexuous, stipes branched once or twice, monochasial, monoverticillate or irregularly biverticillate, with 1-septate at base. Conidiogenous cells monophialidic, hyaline, with conspicuous periclinal thickening. Conidia pale olivaceous-green, solitary, smooth-walled, clavate to oblong-ellipsoidal, slightly curved, with a rounded apical cell, and somewhat conical basal cell, lacking a differentiated foot, 0–3(–4)-septate: 0–1-septate conidia accounting for 8%, 43–65 × 4.5–5.5 μm; 2–3-septate conidia mostly abundant, accounting for 90%, 40.5–67.5 × 4–5.5 μm; 4-septate conidia rare, accounting for 2%, 50.5–57 × 5–5.5 μm. Chlamydospores 5.5–7.5 × 7–8.5 μm, ellipsoidal, pale olivaceous-green, terminal or intercalary, solitary or in chain.

Culture characteristics

Colonies growing on PDA and CMA after 20 days of incubation at 28 °C. Colony on PDA slow-growing, surface thick, rosy buff to white, reverse white, raised, aerial hyphae abundance, reaching 30–33 mm diam., entire margin. Colonies on CMA flat, surface white, reverse translucent to pale white, aerial hyphae sparsely developed, reaching 40–42 mm diam.

Additional materials examined

China • Yunnan province, Huize county; isolated from soils in karst rocky desertification area; Oct 2020; Z.F.Yu; living cultures H77, H102.

Notes

Phylogenetically, the three strains of Atractium yunnanense (YMF 1.06524, H77 and H102) clustered together in a single clade with a high statistical support (BP/BS = 1.00/100) (Fig. 1). The clade containing A. yunnanense and A. crassum makes a sister clade to A. stilbaster. Morphologically, A. yunnanense is similar to A. crassum in having clavate to oblong-ellipsoidal and slightly curved conidia. However, the 2–3-septate conidia are most common in A. yunnanense, whereas A. crassum has most 3–5-septate conidia and no aseptate conidia (Gräfenhan et al. 2011). In addition, the chlamydospores of A. crassum are larger and rounder than A. yunnanense (7–12 μm diam. vs. 5.5–7.5 × 7–8.5 μm diam.). In a comparison of ITS, LSU and tub2 sequences, A. yunnanense (type strain YMF 1.06524) has 97% (546/564 bp, 3 gaps), 98% (857/876 bp, 4 gaps) and 95% (403/419 bp, 3 gaps) similarity to A. crassum (CBS 180.31), respectively. Moreover, the type species A. stilbaster can be morphologically distinguished from A. yunnanense in having smaller conidia, 20–40 × 1.5–2.5 μm vs. 40.5–67.5 × 4–5.5 μm (Seifert 1985).

Nalanthamala Subram., J. Indian bot. Soc. 35: 478 (1956)

Nalanthamala xishuangbannaensis H. Zheng & Z.F. Yu, sp. nov.

MycoBank No: MB854386
Fig. 3

Etymology

Named after the location Xishuangbanna, where the holotype was collected.

Figure 3. 

Nalanthamala xishuangbannaensis (YMF 1.05062, holotype) A colony on PDA after 3 days of inoculation at 28 °C B colony on CMA after 3 days of inoculation at 28 °C C conidia arranged in long chains under low power microscopy D, E acremonium-like conidiophores F–H penicillate conidiophores I conidia. Scale bars: 1.0 cm (A, B); 50 µm (C); 10 µm (D–I).

Type

China • Yunnan province, Xishuangbanna Dai Autonomous Prefecture; isolated from air in forest; Apr 2017; Z.F.Yu, preserved by lyophilization (a metabolically inactive state) in State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan (YMF 1.05062, holotype); ex-type living culture: CGMCC 3.27596.

Description

Sexual morph not observed. Asexual morph on CMA. Hyphae 1.5–3.5 μm wide, hyaline, septate, smooth-walled, branched. Conidiophores hyaline, dimorphic, acremonium-like or penicillium-like. Acremonium-like conidiophores unbranched; phialides cylindrical or slightly tapering toward the tip, straight to slightly bent, 18–37 μm long or longer, 2.3–3.2 μm wide at base, and 1.2–2 μm wide at tip. Penicillium-like conidiophores solitary to aggregated; stipe hyaline, smooth, subcylindrical, septate, 20–80 μm long or longer, up to 3 μm wide at the base; primary and secondary branches subcylindrical, hyaline, smooth, aseptate, 8–16.8 × 2.3–3 µm; phialides cylindrical, flask-shaped, 12.3–22.2 × 2.7–3.2 μm. Conidia either obovate, with an obtuse tip and a truncated base, typically held in long and dry chains, 4–4.8 × 3–3.8 μm or ellipsoidal, cylindrical, or fusiform, with obtuse ends, or with an obtuse tip and a visible, slightly laterally displaced hilum, 4.4–6 × 2.7–3.5 μm. Chlamydospores not observed.

Culture characteristics

Colonies growing on PDA and CMA after 3 days of incubation at 28 °C. Colony on PDA rapid-growing, surface dusty to fine powdery, white to iron gray due to occurrence of conidial masses, reverse white, aerial hyphae flocculent, reaching 53–57 mm diam., indistinct margin. Colony on CMA thin, surface white, reverse translucent, aerial hyphae sparsely developed, reaching 35–37 mm diam.

Additional materials examined

China • Yunnan province, Xishuangbanna Dai Autonomous Prefecture; isolated from air in forest; Apr 2017; Z.F.Yu; living cultures B413, B425.

Notes

Morphologically, Nalanthamala xishuangbannaensis fits well with the generic concepts of Nalanthamala in having acremonium-like or penicillium-like conidiophores and oval or lenticular conidia arranged in chains (Schroers et al. 2005). In the phylogenetic analysis, the three newly obtained strains (YMF 1.05062, B413 and B425) clustered together in a single clade with a strong statistical support (BP/BS = 1.00/100), and the clade was closely related to N. diospyri and N. psidii (Fig. 1). N. xishuangbannaensis can be distinguished from N. diospyri by its wider obovate conidia (3–3.8 μm vs. 2.5 μm in width) and shorter ellipsoidal, cylindrical or fusiform conidia (4.4–6 μm vs. 8–12 μm in length) (Schroers et al. 2005). In a comparison of ITS, LSU and tub2 sequences, N. xishuangbannaensis (type strain YMF 1.05062) has 95% (469/496 bp, 5 gaps), 97% (691/709 bp, 2 gaps) and 91% (294/323 bp, 3 gaps) similarity to N. diospyri (CBS 430.89), respectively. In addition, N. xishuangbannaensis differs from N. psidii in smaller and wider obovate conidia (4–4.8 × 3–3.8 μm vs. 4.5–5.1 × 2.4–2.7 μm) and shorter and wider ellipsoidal, cylindrical or fusiform conidia (4.4–6 × 2.7–3.5 μm vs. 6–11 × 1.7–2.8 μm) (Schroers et al. 2005). Sequences of N. xishuangbannaensis (type strain YMF 1.05062) have 96% similarity in ITS (471/493 bp, 5 gaps) and tub2 (329/351 bp, 4 gaps) to N. psidii (CBS 687.97).

Discussion

Yunnan is uniquely situated at the confluence of three climatic zones: the eastern Asian monsoon zone, the Tibetan Plateau zone, and the tropical monsoon zones of southern Asia and Indochina (Yang et al. 2004). This diverse climatic environment and unique geographical position make the province one of the richest sources of fungi, covering over 40% of the known species in China (Feng and Yang 2018). Nectriaceae species are widespread worldwide, but only one Atractium species (A. fusiformis) and one Nalanthamala species (N. psidii) have been recorded from China (Schroers et al. 2005; Gräfenhan et al. 2011; Perera et al. 2023). Therefore, the proposed two new species further showed Yunnan’s high fungi diversity.

Most of Nectriaceae species are soil-borne saprobes or plant pathogens (Lombard et al. 2015; Perera et al. 2023; Chen et al. 2024). Nonetheless, Atractium species are commonly associated with water (Gräfenhan et al. 2011). For instance, A. crassum was isolated from drinking water in Germany (Gräfenhan et al. 2011), and the three species A. holubovae, A. aquatica and A. fusiformis have been found on submerged decaying wood in the Philippines, France and China respectively (Seifert et al. 1995; Lechat and Fournier 2015; Bao et al. 2023). Only the type species A. stilbaster was discovered on bark or stump in Canada and Germany (Gräfenhan et al. 2011). Significantly, our proposed species A. yunnanense was discovered from soils in a karst rocky desertification area of Yunnan. This finding extends the habitat of Atractium species.

Currently, five species are accepted in Nalanthamala (Wijayawardene et al. 2020; Crous et al. 2021). The type species N. madreeya was isolated from dead stems in India; unfortunately, the type specimen could not be found in the herbaria and the species has not been recollected. Another four Nalanthamala species were reported as associated with wilt and blight diseases from several economically important crops and were more widespread (Schroers et al. 2005). N. psidii causes a destructive wilt disease to Psidium guajava trees and is distributed in Malaysia, South Africa, and Taiwan, possibly restricted to subtropical or tropical regions. N. vermoesenii could cause necrosis and blight to various Arecaceae species and is distributed in Australia, the Czech Republic, Spain and the Unites States, particularly known from warm temperate, Mediterranean, or (sub)tropical climates. N. diospyri could cause a destructive wilt to Diospyros virginiana and is distributed in the United States. Additionally, N. olivacea was isolated from palm in Mexico, Costa Rica, and the Philippines, possibly restricted to tropical regions. Overall, Nalanthamala species are mainly distributed in the warm temperate to tropical regions (Schroers et al. 2005). Our proposed new species N. xishuangbannaensis was isolated from the air in Xishuangbanna of Yunnan, which is a tropical region, and is also the first Nalanthamala species obtained from air.

Despite having cosmopolitan distribution of Nectriaceae species, Atractium and Nalanthamala species have fewer records in China. Significantly, A. fusiformis and A. yunnanense were first discovered from Yunnan (Bao et al. 2023). Two Nalanthamala species, N. psidii and N. xishuangbannaensis, were found in China. N. psidii, as a widespread species, was found in Taiwan (Rossman et al. 2013), and N. xishuangbannaensis was first reported from Yunnan. The discovery of these Atractium and Nalanthamala species in Yunnan proves once again that it is rich in fungal species. Further studies are needed to understand the pathogenicity of the newly discovered Atractium and Nalanthamala species.

Acknowledgments

This work acknowledges the Microbial Library of the Germplasm Bank of wild species from Southwest China for the preservation of strains.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This work was financed by the National Natural Science Foundation Program of PR China (32170017, 32370017).

Author contributions

YZ designed research; ZH and DX performed experiments; ZH wrote the original draft; YZ and LH reviewed and edited the manuscript. All authors have read and approved the final manuscript.

Author ORCIDs

Hua Zheng https://orcid.org/0000-0002-0794-6742

Xinwen Dai https://orcid.org/0009-0000-1175-8456

Haiyan Li https://orcid.org/0000-0002-7041-3199

Zefen Yu https://orcid.org/0000-0002-6246-5861

Data availability

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

References

  • Bao DF, Hyde KD, Maharachchikumbura SS, Perera RH, Thiyagaraja V, Hongsanan S, Wanasinghe DN, Shen H-W, Tian X-G, Yang L-Q, Nalumpang S, Luo Z-L (2023) Taxonomy, phylogeny and evolution of freshwater Hypocreomycetidae (Sordariomycetes). Fungal Diversity 121(1): 1–94. https://doi.org/10.1007/s13225-023-00521-8
  • Chang DC, Grant GB, O’Donnell K, Wannemuehler KA, Noble-Wang J, Rao CY, Jacobson LM, Crowell CS, Sneed RS, Lewis FMT, Schaffzin JK, Kainer MA, Genese CA, Alfonso EC, Jones DB, Srinivasan A, Fridkin SK, Park BJ, Fusarium Keratitis Investigation Team (2006) Multistate outbreak of Fusarium keratitis associated with use of a contact lens solution. Journal of the American Medical Association 296(8): 953–963. https://doi.org/10.1001/jama.296.8.953
  • Chaverri P, Salgado C, Hirooka Y, Rossman A, Samuels G (2011) Delimitation of Neonectria and Cylindrocarpon (Nectriaceae, Hypocreales, Ascomycota) and related genera with Cylindrocarpon-like anamorphs. Studies in Mycology 68(1): 57–78. https://doi.org/10.3114/sim.2011.68.03
  • Chen H, White JF, Malik K, Li C (2024) Molecular assessment of oat head blight fungus, including a new genus and species in a family of Nectriaceae. International Journal of Food Microbiology 417: 110715. https://doi.org/10.1016/j.ijfoodmicro.2024.110715
  • Crous PW, Osieck ER, Jurjević Ž, Boers J, Van Iperen A, Starink-Willemse M, Dima B, Balashov S, Bulgakov TS, Johnston PR, Morozova OV, Pinruan U, Sommai S, Alvarado P, Decock CA, Lebel T, McMullan-Fisher S, Moreno G, Shivas RG, Zhao L, Abdollahzadeh J, Abrinbana M, Ageev DV, Akhmetova G, Alexandrova AV, Altés A, Amaral AGG, Angelini C, Antonín V, Arenas F, Asselman P, Badali F, Baghela A, Bañares A, Barreto RW, Baseia IG, Bellanger J-M, Berraf-Tebbal A, Biketova A Yu, Bukharova NV, Burgess TI, Cabero J, Câmara MPS, Cano-Lira JF, Ceryngier P, Chávez R, Cowan DA, de Lima AF, Oliveira RL, Denman S, Dang QN, Dovana F, Duarte IG, Eichmeier A, Erhard A, Esteve-Raventós F, Fellin A, Ferisin G, Ferreira RJ, Ferrer A, Finy P, Gaya E, Geering ADW, Gil-Durán C, Glässnerová K, Glushakova AM, Gramaje D, Guard FE, Guarnizo AL, Haelewaters D, Halling RE, Hill R, Hirooka Y, Hubka V, Iliushin VA, Ivanova DD, Ivanushkina NE, Jangsantear P, Justo A, Kachalkin AV, Kato S, Khamsuntorn P, Kirtsideli IY, Knapp DG, Kochkina GA, Koukol O, Kovács GM, Kruse J, Kumar TKA, Kušan I, Læssøe T, Larsson E, Lebeuf R, Levicán G, Loizides M, Marinho P, Luangsa-ard JJ, Lukina EG, Magaña-Dueñas V, Maggs-Kölling G, Malysheva EF, Malysheva VF, Martín B, Martín MP, Matočec N, McTaggart AR, Mehrabi-Koushki M, Mešić A, Miller AN, Mironova P, Moreau P-A, Morte A, Müller K, Nagy LG, Nanu S, Navarro-Ródenas A, Nel WJ, Nguyen TH, Nóbrega TF, Noordeloos ME, Olariaga I, Overton BE, Ozerskaya SM, Palani P, Pancorbo F, Papp V, Pawłowska J, Pham TQ, Phosri C, Popov ES, Portugal A, Pošta A, Reschke K, Reul M, Ricci GM, Rodríguez A, Romanowski J, Ruchikachorn N, Saar I, Safi A, Sakolrak B, Salzmann F, Sandoval-Denis M, Sangwichein E, Sanhueza L, Sato T, Sastoque A, Senn-Irlet B, Shibata A, Siepe K, Somrithipol S, Spetik M, Sridhar P, Stchigel AM, Stuskova K, Suwannasai N, Tan YP, Thangavel R, Tiago I, Tiwari S, Tkalčec Z, Tomashevskaya MA, Tonegawa C, Tran HX, Tran NT, Trovão J, Trubitsyn VE, Van Wyk J, Vieira WAS, Vila J, Visagie CM, Vizzini A, Volobuev SV, Vu DT, Wangsawat N, Yaguchi T, Ercole E, Ferreira BW, de Souza AP, Vieira BS, Groenewald JZ (2021) Fungal Planet description sheets: 1284–1382. Persoonia 47(1): 178–374. https://doi.org/10.3767/persoonia.2021.47.06
  • Dennis RWG (1960) British cup fungi and their allies: An introduction to the Ascomycetes. Bernard Quaritch Ltd., London, 280 pp.
  • Gräfenhan T, Schroers HJ, Nirenberg H, Seifert K (2011) An overview of the taxonomy, phylogeny, and typification of nectriaceous fungi in Cosmospora, Acremonium, Fusarium, Stilbella, and Volutella. Studies in Mycology 68(1): 79–113. https://doi.org/10.3114/sim.2011.68.04
  • Guarro J (2013) Fusariosis, a complex infection caused by a high diversity of fungal species refractory to treatment. European Journal of Clinical Microbiology & Infectious Diseases 32(12): 1491–1500. https://doi.org/10.1007/s10096-013-1924-7
  • 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.
  • Hoenigl M, Salmanton-García J, Walsh TJ, Nucci M, Neoh CF, Jenks JD, Lackner M, Sprute R, Al-Hatmi AMS, Bassetti M, Carlesse F, Freiberger T, Koehler P, Lehrnbecher T, Kumar A, Prattes J, Richardson M, Revankar S, Slavin MA, Stemler J, Spiess B, Taj-Aldeen SJ, Warris A, Woo PCY, Young J-AH, Albus K, Arenz D, Arsic-Arsenijevic V, Bouchara J-P, Chinniah TR, Chowdhary A, de Hoog GS, Dimopoulos G, Duarte RF, Hamal P, Meis JF, Mfinanga S, Queiroz-Telles F, Patterson TF, Rahav G, Rogers TR, Rotstein C, Wahyuningsih R, Seidel D, Cornely OA (2021) Global guideline for the diagnosis and management of rare mould infections: An initiative of the European Confederation of Medical Mycology in cooperation with the International Society for Human and Animal Mycology and the American Society for Microbiology. The Lancet, Infectious Diseases 21(8): e246–e257. https://doi.org/10.1016/S1473-3099(20)30784-2
  • Jayawardena RS, Hyde KD, Jeewon R, Ghobad-Nejhad M, Wanasinghe DN, Liu NG, Phillips AJL, Oliveira-Filho JRC, da Silva GA, Gibertoni TB, Abeywikrama P, Carris LM, Chethana KWT, Dissanayake AJ, Hongsanan S, Jayasiri SC, McTaggart AR, Perera RH, Phutthacharoen K, Savchenko KG, Shivas RG, Thongklang N, Dong W, Wei DP, Wijayawardena NN, Kang J-C (2019a) One stop shop II: taxonomic update with molecular phylogeny for important phytopathogenic genera: 26–50 (2019). Fungal Diversity 94: 41–129. https://doi.org/10.1007/s13225-019-00418-5
  • Jayawardena RS, Hyde KD, McKenzie EH, Jeewon R, Phillips AJ, Perera RH, de Silva NI, Maharachchikumburua SSN, Samarakoon MC, Ekanayake AH, Tennakoon DS, Dissanayake AJ, Norphanphoun C, Lin C, Manawasinghe IS, Tian Q, Brahmanage R, Chomnunti P, Hongsanan S, Jayasiri SC, Halleen F, Bhunjun CS, Karunarathna A, Wang Y (2019b) One stop shop III: taxonomic update with molecular phylogeny for important phytopathogenic genera: 51–75 (2019). Fungal Diversity 98: 77–160. https://doi.org/10.1007/s13225-019-00433-6
  • Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35(6): 1547–1549. https://doi.org/10.1093/molbev/msy096
  • Lechat C, Fournier J (2015) Varicosporella, a new aquatic genus in the Nectriaceae from France. Ascomycete. Org: Revue Internationale pour la Taxinomie des Ascomycota 7(1): 1–8. https://doi.org/10.25664/art-0121
  • Link HF (1809) Observationes in ordines plantarum naturales. Dissertatio I Magazin der Gesellschaft Naturforschenden Freunde Berlin 3: 3–42.
  • Lv R, Yang X, Qiao M, Fang L, Li J, Yu Z (2022) Exophiala yunnanensis and Exophiala yuxiensis (Chaetothyriales, Herpotrichiellaceae), two new species of soil-inhabiting Exophiala from Yunnan Province, China. MycoKeys 94: 109–124. https://doi.org/10.3897/mycokeys.94.96782
  • Mi X, Feng G, Hu Y, Zhang J, Chen L, Corlett RT, Hughes AC, Pimm S, Schmid B, Shi S, Svenning J-C, Ma K (2021) The global significance of biodiversity science in China: An overview. National Science Review 8(7): nwab032. https://doi.org/10.1093/nsr/nwab032
  • Myers N, Mittermeier RA, Mittermeier CG, Da Fonseca GA, Kent J (2000) Biodiversity hotspots for conservation priorities. Nature 403(6772): 853–858. https://doi.org/10.1038/35002501
  • Orme CDL, Davies RG, Burgess M, Eigenbrod F, Pickup N, Olson VA, Webster AJ, Ding T-S, Rasmussen PC, Ridgely RS, Stattersfield AJ, Bennett PM, Blackburn TM, Gaston KJ, Owens IPF (2005) Global hotspots of species richness are not congruent with endemism or threat. Nature 436(7053): 1016–1019. https://doi.org/10.1038/nature03850
  • Perera R, Hyde K, Jones E, Maharachchikumbura S, Bundhun D, Camporesi E, Akulov A, Liu JK, Liu ZY (2023) Profile of Bionectriaceae, Calcarisporiaceae, Hypocreaceae, Nectriaceae, Tilachlidiaceae, Ijuhyaceae fam. nov., Stromatonectriaceae fam. nov. and Xanthonectriaceae fam. nov. Fungal Diversity 118(1): 95–271. https://doi.org/10.1007/s13225-022-00512-1
  • Qiao M, Zheng H, Lv R, Yu Z (2020) Neodactylariales, Neodactylariaceae (Dothideomycetes, Ascomycota): New order and family, with a new species from China. MycoKeys 73(6): 69–85. https://doi.org/10.3897/mycokeys.73.54054
  • Qiao M, Zheng H, Guo J-S, F. Castañeda-Ruiz R, Xu J-P, Peng J, Zhang K-Q, Yu Z-F (2021) Two new asexual genera and six new asexual species in the family Microthyriaceae (Dothideomycetes, Ascomycota) from China. MycoKeys 85: 1–30. https://doi.org/10.3897/mycokeys.85.70829
  • 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
  • Rossman AY, Samuels GJ, Rogerson CT, Lowen R (1999) Genera of Bionectriaceae, Hypocreaceae and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42(42): 1–248.
  • Rossman AY, Seifert KA, Samuels GJ, Minnis AM, Schroers HJ, Lombard L, Crous PW, Põldmaa K, Cannon PF, Summerbell RC, Geiser DM, Zhuang W, Hirooka Y, Herrera C, Salgado-Salazar C, Chaverri P (2013) Genera in Bionectriaceae, Hypocreaceae, and Nectriaceae (Hypocreales) proposed for acceptance or rejection. IMA Fungus 4(1): 41–51. https://doi.org/10.5598/imafungus.2013.04.01.05
  • Schroers HJ, Geldenhuis M, Wingfield M, Schoeman M, Yen YF, Shen WC, Wingfield BD (2005) Classification of the guava wilt fungus Myxosporium psidii, the palm pathogen Gliocladium vermoesenii and the persimmon wilt fungus Acremonium diospyri in Nalanthamala. Mycologia 97(2): 375–395. https://doi.org/10.1080/15572536.2006.11832814
  • Seifert KA, Stanley SJ, Hyde KD (1995) Stilbella holubovae, a new synnematous hyphomycete species on driftwood from the Philippines and South Africa. Sydowia 47: 258–262.
  • Subramanian CV (1956) Hyphomycetes—II. Journal of the Indian Botanical Society 35: 446–494.
  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin J, Higgins DG (1997) The CLUSTAL X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24): 4876–4882. https://doi.org/10.1093/nar/25.24.4876
  • Tulasne L, Tulasne C (1865) Selecta Fungorum Carpologia: Nectriei-Phacidiei-Pezizei, 3.
  • Turner D, Kovacs W, Kuhls K, Lieckfeldt E, Peter B, Arisan-Atac I, Strauss J, Samuels GJ, Börner T, Kubicek CP (1997) Biogeography and phenotypic variation in Trichoderma sect. Longibrachiatum and associated Hypocrea species. Mycological Research 101(4): 449–459. https://doi.org/10.1017/S0953756296002845
  • 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
  • Vincent AL, Cabrero JE, Greene JN, Sandin RL (2003) Successful voriconazole therapy of disseminated Fusarium solani in the brain of a neutropenic cancer patient. Cancer Control 10(5): 414–419. https://doi.org/10.1177/107327480301000511
  • Wijayawardene NN, Hyde KD, Al-Ani LKT, Tedersoo L, Haelewaters D, Rajeshkumar KC, Zhao RL, Aptroot A, Leontyev DV, Saxena RK, Tokarev YS, Dai DQ, Letcher PM, Stephenson SL, Ertz D, Lumbsch HT, Kukwa M, Issi IV, Madrid H, Phillips AJL, Selbmann L, Pfliegler WP, Horváth E, Bensch K, Kirk PM, Kolaříková K, Raja HA, Radek R, Papp V, Dima B, Ma J, Malosso E, Takamatsu S, Rambold G, Gannibal PB, Triebel D, Gautam AK, Avasthi S, Suetrong S, Timdal E, Fryar SC, Delgado G, Réblová M, Doilom M, Dolatabadi S, Pawłowska J, Humber RA, Kodsueb R, Sánchez-Castro I, Goto BT, Silva DKA, de Souza FA, Oehl F, da Silva GA, Silva IR, Błaszkowski J, Jobim K, Maia LC, Barbosa FR, Fiuza PO, Divakar PK, Shenoy BD, Castañeda-Ruiz RF, Somrithipol S, Lateef AA, Karunarathna SC, Tibpromma S, Mortimer PE, Wanasinghe DN, Phookamsak R, Xu J, Wang Y, Tian F, Alvarado P, Li DW, Kušan I, Matočec N, Maharachchikumbura SSN, Papizadeh M, Heredia G, Wartchow F, Bakhshi M, Boehm E, Youssef N, Hustad VP, Lawrey JD, Santiago ALCMA, Bezerra JDP, Souza-Motta CM, Firmino AL, Tian Q, Houbraken J, Hongsanan S, Tanaka K, Dissanayake AJ, Monteiro JS, Grossart HP, Suija A, Weerakoon G, Etayo J, Tsurykau A, Vázquez V, Mungai P, Damm U, Li QR, Zhang H, Boonmee S, Lu YZ, Becerra AG, Kendrick B, Brearley FQ, Motiejūnaitė J, Sharma B, Khare R, Gaikwad S, Wijesundara DSA, Tang LZ, He MQ, Flakus A, Rodriguez-Flakus P, Zhurbenko MP, McKenzie EHC, Stadler M, Bhat DJ, Liu JK, Raza M, Jeewon R, Nassonova ES, Prieto M, Jayalal RGU, Erdoğdu M, Yurkov A, Schnittler M, Shchepin ON, Novozhilov YK, Silva-Filho AGS, Liu P, Cavender JC, Kang Y, Mohammad S, Zhang LF, Xu RF, Li YM, Dayarathne MC, Ekanayaka AH, Wen TC, Deng CY, Pereira OL, Navathe S, Hawksworth DL, Fan XL, Dissanayake LS, Kuhnert E, Grossart HP, Thines M (2020) Outline of Fungi and fungi-like taxa. Mycosphere: Journal of Fungal Biology 11(1): 1160–1456. https://doi.org/10.5943/mycosphere/11/1/8
  • Woudenberg J, Aveskamp M, De Gruyter J, Spiers A, Crous PW (2009) Multiple Didymella teleomorphs are linked to the Phoma clematidina morphotype. Persoonia 22(1): 56–62. https://doi.org/10.3767/003158509X427808
  • Yu Z, Jiang X, Zheng H, Zhang H, Qiao M (2022) Fourteen new species of foliar Colletotrichum associated with the invasive plant Ageratina adenophora and surrounding crops. Journal of Fungi (Basel, Switzerland) 8(2): 185. https://doi.org/10.3390/jof8020185
  • Zheng H, Zhang Z, Liu DZ, Yu ZF (2019) Memnoniella sinensis sp. nov., a new species from China and a key to species of the genus. International Journal of Systematic and Evolutionary Microbiology 69(10): 3161–3169. https://doi.org/10.1099/ijsem.0.003605
  • Zheng H, Yang XQ, Deng JS, Xu JP, Yu ZF (2020) Beltrania sinensis sp. nov., an endophytic fungus from China and a key to species of the genus. International Journal of Systematic and Evolutionary Microbiology 70(2): 1178–1185. https://doi.org/10.1099/ijsem.0.003897
  • Zheng H, Qiao M, Lv Y, Du X, Zhang KQ, Yu Z (2021a) New species of Trichoderma isolated as endophytes and saprobes from Southwest China. Journal of Fungi (Basel, Switzerland) 7(6): 467. https://doi.org/10.3390/jof7060467
  • Zheng H, Qiao M, Xu J, Yu Z (2021b) Culture-based and culture-independent assessments of endophytic fungal diversity in aquatic plants in Southwest China. Frontiers in Fungal Biology 2: 692549. https://doi.org/10.3389/ffunb.2021.692549
  • Zheng H, Yu Z, Jiang X, Fang L, Qiao M (2022) Endophytic Colletotrichum species from aquatic plants in Southwest China. Journal of Fungi (Basel, Switzerland) 8(1): 87. https://doi.org/10.3390/jof8010087
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