Diversity and Distribution of Colletotrichum Species Causing Anthracnose in China

Abstract

This study conducted a systematic investigation and identification of pathogenic fungi associated with anthracnose symptoms on economically important plants across multiple provinces in China (Beijing, Fujian, Guangdong, Guizhou, and Shaanxi). Through multi-locus phylogenetic analysis (ITS, gapdh, chs-1, act, tub2, his3, and cal) and morphological characterization of 67 strains, a total of 16 Colletotrichum species were identified, belonging to six species complexes (C. acutatum, C. boninense, C. destructivum, C. gloeosporioides, C. orchidearum, and C. spaethianum). Among these, four novel species were described: Colletotrichum aquilariae, C. crataegi, C. dongguanense, and C. flavosporum. The study also confirmed 12 known species: C. boninense, C. fioriniae, C. fructicola, C. godetiae (with C. americanum proposed as its synonym), C. gloeosporioides (with C. juglandicola, C. juglandium, and C. peakense proposed as its synonyms), C. karsti, C. nymphaeae, C. orchidearum (with C. subplurivorum proposed as its synonym), C. plurivorum, C. siamense, C. sojae, and C. spaethianum. The research revealed significant pathogen species diversity, distinct geographical distribution patterns (greatest diversity in Beijing, novel species primarily from Guangdong), and host preferences (e.g., C. gloeosporioides was the most widely distributed and dominant species on walnut). Furthermore, ten new host records were reported. The study explored correlations between pathogens and their hosts, particularly walnut, providing a crucial foundation for understanding the pathogen composition and ecology of anthracnose diseases affecting plants in China.

1. Introduction

Colletotrichum (Glomerellaceae) is distributed as one of the top ten global phytopathogenic genera worldwide [1]. Colletotrichum fungus grows and reproduces more rapidly in tropical and subtropical regions [2]. They can infect the aerial tissues of plants [3]. The typical symptoms are forming light to dark brown spots, which are usually round or irregular in shape [4,5]. In severe cases, they can merge into large areas, leading to extensive necrosis and wilting of plant leaves, stems, and other parts [6,7]. In addition, Colletotrichum is also capable of inducing post-harvest rot, which causes fruits and vegetables to emit a foul odor [8].

The morphological characteristics of Colletotrichum species exhibit variation and overlap. The conidia of the C. acutatum species complex are mostly long-elliptical or long-ovoid and relatively large [9]. The length-to-width ratio and the distinct scar at the base of the conidia of C. boninense show subtle differences compared to C. gloeosporioides [10,11]. The conidia of the C. gloeosporioides species complex are mostly long-elliptical or cylindrical and relatively small [12]. Therefore, accurate species identification is difficult based solely on morphological characteristics. Such morphological ambiguities highlight the need for complementary molecular or biochemical approaches to ensure reliable species delimitation.

With the development and refinement of molecular biological techniques, the classification and species concepts of Colletotrichum have become increasingly well-defined [13,14,15,16]. Jayawardena et al. [14] have provided molecular data for 248 species, divided into 14 species complexes and 13 singleton species. Subsequently, Liu et al. [16] have provided molecular data for 280 species, divided into 16 species complexes and 15 singleton species of Colletotrichum. Colletotrichum karsti, C. siamense, C. fructicola, C. fioriniae, and C. gloeosporioides are species with high isolation rates in China [16], primarily infecting woody plants such as walnut, Camellia oleifera, and Cinnamomum camphora [2].

Multiple distinct Colletotrichum species can infect the same host. Existing research indicates that walnut anthracnose can be caused by several groups, including the Colletotrichum acutatum species complex, the C. boninense species complex, and the C. gloeosporioides species complex [17,18,19,20,21]. De Silva et al. [22] report different Colletotrichum species causing anthracnose in chili peppers. These studies indicate that walnuts, Camellia oleifera, and chili peppers are more susceptible to infection by Colletotrichum fungi compared to other plants.

The aims of this study are to (1) systematically identify the species composition of anthracnose fungi on walnuts and determine whether these species differ in their geographical distribution and host adaptation and (2) expand the investigation to non-walnut hosts, exploring whether ecological isolation or coevolution exists between these hosts and the anthracnose fungi predominantly occurring on walnuts and reveal the pathogen diversity of the hosts.

2. Materials and Methods

2.1. Disease Survey and Sample Isolations

From 2023 to 2024, a total of 110 symptomatic samples of branch cankers and leaf spots were collected from five provincial regions in China (Beijing, Fujian, Guangdong, Guizhou, and Shaanxi), among which 67 samples (Figure 1) exhibited typical anthracnose symptoms. The disease samples were placed in specimen bags, and detailed sample information was recorded. They were then brought back to the laboratory for isolation. The tissue isolation method was used to isolate the leaf spot samples [23]. The fresh fruiting bodies on the branch cankers were cut open with a sterilized knife to expose the spore masses. The spore masses were transferred to PDA plates with a sterilized needle to obtain pure cultures at 25 °C. Obtain the sexual and asexual morphs of fungi on PDA at 25 °C. Capture microscopic images of fungi using a DIC microscope. All samples have been deposited in the Museum of the Beijing Forestry University (BJFC) and the Institute of Microbiology, Chinese Academy of Sciences (IMCAS). The pure cultures obtained have been deposited in the China Forestry Culture Collection Center (CFCC, https://cfcc.caf.ac.cn/) and the Laboratory of Forest Pathology Resources (LFPR), Beijing Forestry University.

2.2. DNA Extraction and PCR Amplification

Genomic DNA was extracted from cultures growing on a PDA medium using a modified CTAB method [24]. To identify the strains, we amplified the following gene loci: the internal transcribed spacer (ITS) regions, glyceraldehyde 3-phosphate dehydrogenase (gapdh), partial sequences of the chitin synthase 1 (chs-1), actin (act), β-tubulin 2 (tub2), Histone3 (his3), and Calmodulin (cal) [2,16]. We used the respective primer pairs: ITS1/ITS4, GDF1/GDR1, CHS-79F/CHS-345R, ACT-512F/ACT-783R, T1/Bt2b, CYLH3F/ CYLH3R, and CL1C/CL2C [12,16,25,26,27,28,29,30]. The PCR conditions for different genes used in the genus Colletotrichum are listed in

Table 1. Gene fragments and the PCR thermal cycle program used in this study.

Locus	PCR Primers	PCR: Thermal Cycles: (Annealing Temp. in Bold)	Reference
ITS	ITS1/ITS4	(95 °C: 30 s, 51 °C: 30 s, 72 °C: 1 min) × 35 cycles	[25]
gapdh	GDF1/GDR1	(95 °C: 30 s, 58 °C: 30 s, 72 °C: 1 min) × 35 cycles	[26]
chs-1	CHS-79F/CHS-345R	(95 °C: 30 s, 58 °C: 30 s, 72 °C: 1 min) × 35 cycles	[27]
act	ACT-512F/ACT-783R	(95 °C: 45 s, 55 °C: 45 s, 72 °C: 1 min) × 35 cycles	[28]
tub2	T1/Bt-2b	(95 °C: 30 s, 55 °C: 30 s, 72 °C: 1 min) × 35 cycles	[29]
his3	CYLH3F/ CYLH3R	(95 °C: 30 s, 58 °C: 30 s, 72 °C: 1 min) × 35 cycles	[30]
cal	CL1C/CL2C	(95 °C: 30 s, 54 °C: 20 s, 72 °C: 1 min) × 35 cycles	[12,16]

. The amplicons were sequenced by the SinoGenoMax Company Limited (Beijing, China). All sequence data obtained in this study have been submitted to GenBank (Supplementary Table S1).

2.3. Phylogenetic Analysis

Colletotrichum species sequences were downloaded from NCBI for phylogenetic analysis, referring to Zhang and Chen et al. [31], Zhang and Nizamani et al. [7], Sui et al. [2], and Li et al. [32]. Sequence assembly was performed using Seqman v. 7.1.0 software. Alignment was conducted using MAFFT v. 7 [33] (http://mafft.cbrc.jp/alignment/server/index.html, accessed on 7 October 2025). Sequence trimming and editing were carried out using MEGA v. 6 [34]. Phylogenetic analysis was performed using maximum likelihood (ML) and Bayesian inference (BI). PhyML v. 3.0 was used for ML analysis, with the GTR nucleotide substitution model and the bootstrap (BS) method with 1000 replicates [35]. Bayesian Inference (BI) analysis was performed using the Markov chain Monte Carlo (MCMC) algorithm [36]. Two MCMC chains were run starting from a random tree for 1,000,000 generations, with trees saved every 1000 generations and the first 25% of trees discarded as burn-in for posterior probability (BPP). The BPP of each analysis was used to evaluate the remaining trees. The phylogenetic tree was visualized in FigTree v.1.3.1 [37].

2.4. Correlation Analysis Between Pathogens and Hosts

Correlation analysis between pathogens and hosts was performed using Excel 2020 software. A structured database was constructed by integrating species identification results of isolated strains, host information, and geographic origins (provincial/municipal sampling sites). Occurrence frequency and relative abundance of different species were calculated using Excel pivot tables to clarify differences in the composition of dominant species across hosts. The relative abundances of different species in walnut were computed using the pie chart tool in Excel 2020 software to evaluate the dominant species in walnut. Combined with geographic and host information, evolutionary and ecological associations were further elucidated.

3. Results

3.1. Phylogenetic Analyses

Based on the single-gene phylogenetic analysis of the ITS region, it was confirmed that the 67 isolates belong to six species complexes of C. acutatum, C. boninense, C. destructivum, C. gloeosporioides, C. orchidearum, and C. spaethianum. Based on multi-gene phylogenetic analysis (ITS, gapdh, chs-1, act, tub2, his3, and cal), the 67 isolates belonged to four new species and 12 known species within the six species complexes, with the specific results as follows:

3.1.1. Colletotrichum acutatum Species Complex Phylogenetic Analysis

The Colletotrichum acutatum species complex was analyzed using multiple gene loci (ITS, gapdh, chs-1, act, tub2, and his3) (Figure 2). The outgroup was C. orchidophilum (CBS 632.80 and CBS 631.80). In the ML analysis based on the combined gene dataset, the best likelihood value was −11,632.287603. The matrix had 846 distinct alignment patterns. The proportion of gaps and completely undetermined characters was 12.39%. The estimated base frequencies were as follows: A = 0.214646, C = 0.281884, G = 0.266176, T = 0.237294; substitution rates: AC = 0.901509, AG = 3.938106, AT = 1.080309, CG = 0.554337, CT = 4.209148, GT = 1.000000; the shape parameter of the gamma distribution α = 0.247371. The 13 strains in this study formed three distinct clades, with 13 strains clustering with C. fioriniae, C. godetiae, and C. nymphaeae, respectively (Figure 2). All three clades were supported with over 89% ML and 1.00 Bayesian posterior probability (BYPP). The 13 strains were identified as three known species (C. fioriniae, C. godetiae, and C. nymphaeae) within the C. acutatum species complex (Figure 2).

3.1.2. Colletotrichum boninense Species Complex Phylogenetic Analysis

The Colletotrichum boninense species complex was analyzed using multiple gene loci (ITS, gapdh, chs-1, act, tub2, and his3) (Figure 3). Colletotrichum truncatum (CBS 151.35) was used as the outgroup. The best likelihood value of the ML tree was −11,896.599473. The matrix had 939 distinct alignment patterns. The proportion of gaps and completely undetermined characters was 14.06%. The estimated base frequencies were as follows: A = 0.230169, C = 0.259637, G = 0.298228, T = 0.211966; substitution rates: AC = 1.062959, AG = 3.732917, AT = 0.904273, CG = 0.735975, CT = 3.422578, GT = 1.000000; the shape parameter of the gamma distribution α = 0.302366. The C. boninense species complex involved eight strains in this study, which formed four separate clades (Figure 3). The ML values for the three clades were above 90% and the Bayesian posterior probabilities (BYPP) were above 0.9. The six strains were identified as C. boninense, and two strains were identified as C. karsti (Figure 3).

3.1.3. Colletotrichum destructivum Species Complex Phylogenetic Analysis

The Colletotrichum destructivum species complex was analyzed using multiple gene loci (ITS, gapdh, chs-1, act, tub2, and his3) (Figure 4). Colletotrichum coccodes (CBS 369.75) was selected as an outgroup. The best likelihood value for the C. destructivum species complex was −8634.934944. The matrix had 651 distinct alignment patterns. The proportion of gaps and completely undetermined characters was 15.49%. The estimated base frequencies were as follows: A = 0.216431, C = 0.268314, G = 0.287624, T = 0.227631; substitution rates: AC = 1.353115, AG = 5.041429, AT = 1.199580, CG = 0.588826, GT = 1.000000, CT = 4.193986; the shape parameter of the gamma distribution α = 0.253411. Two strains in this study formed a single, separate clade, which was identified as one new species (C. crataegi) within the C. destructivum species complex.

3.1.4. Colletotrichum gloeosporioides Species Complex Phylogenetic Analysis

The Colletotrichum gloeosporioides species complex was analyzed using multiple gene loci (ITS, gapdh, chs-1, act, tub2, and cal) (Figure 5). The outgroup was C. boninense (CBS 123755). The best likelihood value was −21,030.171117. The matrix had 1574 distinct alignment patterns with a 30.93% proportion of gaps and completely undetermined characters. The estimated base frequencies were as follows: A = 0.225648, C = 0.295175, G = 0.251021, T = 0.228155; substitution rates: AC = 1.180654, AG = 3.528904, AT = 1.177066, CG = 0.872057, GT = 1.000000, CT = 3.895772; the shape parameter of the gamma distribution α = 0.404514. 28 strains in this study were identified as three known species (C. fructicola, C. siamense, and C. gloeosporioides) (Figure 5). Six strains were identified as three new species (C. aquilariae, C. dongguanense, and C. flavosporum) within the C. gloeosporioides species complex (Figure 5). The ML values for these eight species were all above 95% and the Bayesian posterior probabilities (BYPP) were all 1.00.

3.1.5. Colletotrichum orchidearum Species Complex Phylogenetic Analysis

The Colletotrichum orchidearum species complex was analyzed using multiple gene loci (ITS, gapdh, chs-1, act, tub2, and his3) (Figure 6). The outgroup was C. magnum (CBS 519.97). The best likelihood value was −5875.108038. The matrix had 434 distinct alignment patterns with an 11.21% proportion of gaps and completely undetermined characters. The estimated base frequencies were as follows: A = 0.212098, C = 0.315800, G = 0.257503, T = 0.214599; substitution rates: AC = 1.108763, AG = 3.317336, AT = 0.795605, CG = 0.686179, GT = 1.000000, CT = 4.883049; the shape parameter of the gamma distribution α = 0.212737. Seven strains in this study were identified as three known species (C. orchidearum, C. plurivorum, and C. sojae) (Figure 6). The ML values for these known species were all above 98% and the Bayesian posterior probabilities (BYPP) were all 1.00.

3.1.6. Colletotrichum spaethianum Species Complex Phylogenetic Analysis

The Colletotrichum spaethianum species complex was analyzed using multiple gene loci (ITS, gapdh, chs-1, act, tub2, and his3) (Figure 7). The outgroup was C. scovillei (CBS 126529). The best likelihood value was −8359.303202. The matrix had 675 distinct alignment patterns with a 17.29% proportion of gaps and completely undetermined characters. The estimated base frequencies were as follows: A = 0.219187, C = 0.303015, G = 0.250183, T = 0.227615; substitution rates: AC = 0.958726, AG = 3.796058, AT = 1.123702, CG = 0.680117, GT = 1.000000, CT = 5.575032; the shape parameter of the gamma distribution α = 0.284355. Three strains in this study were identified as C. spaethianum (Figure 7). The ML values for these known species were all above 100% and the Bayesian posterior probabilities (BYPP) were all 1.00.

3.2. Taxonomy

The 67 strains studied were classified into 16 species, including 4 novel species and 12 known species. Detailed morphological descriptions are provided below for all the species studied in culture. Exceptions were made for the 9 known species C. fioriniae, C. fructicola, C. godetiae, C. karsti, C. nymphaeae, C. orchidearum, C. plurivorum, C. sojae, and C. spaethianum as they have already been described in great detail in the studies by Sui et al. [2], Zhang et al. [7], de Silva et al. [22], Damm et al. [38] and Damm et al. [10], and Damm et al. [39].

Colletotrichum aquilariae W.S. Zhang & X.L. Fan, sp. nov. (Figure 8).

MycoBank: MB859290

Etymology: Named after the host genus of the collected sample, Aquilaria.

Holotype: HMAS 353996

Description: Sexual morph not observed. Asexual morph: Sporulating on PDA. Conidiomata scattered or gregarious, acervular, semi-immersed to immersed, hyaline or dark brown, unbranched, septate. Conidiophores usually reduced to conidiogenous cells, unbranched. Conidiogenous cells hyaline or dark brown, straight or curved, cylindrical or ampulliform, smooth-walled, 8.5–19.5(–47.0) × 1.5–3.1 μm (av. = 12.5 × 2.2 μm, n = 50). Conidia hyaline, cylindrical, aseptate, smooth-walled, no oil droplets, 10.5–12.5 × 3.5–4.8 μm (av. = 11.2 × 4.1 μm, n = 50), L/W ratio = 2.7. Appressoria elliptical to sub-elliptical, hyaline to dark brown, single, 4.5–10.8 × 2.7–8.5 μm (av. = 5.9 × 4.2 μm, n = 20). Setae not observed.

Culture characteristics: Colonies on PDA white, spreading, with the center of colonies having abundant white flocculent aerial mycelium and even margin, reaching a diameter of 60 mm after 8 d at 25 °C.

Typus: CHINA, Guangdong Province, Dongguan City, Xingtang Xinghua Road, collected from diseased leaves of Aquilaria sinensis, 113°44′40″ E, 22°57′33″ N, 7 December 2023, Xinlei Fan (holotype HMAS 353996; ex-holotype culture CFCC 72436); ibid. (living culture CFCC 72437).

Notes: Colletotrichum aquilariae is revealed in the multi-gene phylogram as a distinct clade with full support (BI/ML = 1.00/100) (Figure 5). Colletotrichum aquilariae forms a group close to C. alienum and C. hystricis, but it differs from C. alienum in the nucleotide sequence by 6 bp in the ITS, 4 bp in gapdh, 6 bp in act, 4 bp in tub2, and 5 bp in cal. It can also differ from C. hystricis in the nucleotide sequence by 7 bp in the ITS, 6 bp in gapdh, and 10 bp in act. Morphologically, the conidia of C. aquilariae are smaller than those of C. alienum (10.5–12.5 × 3.5–4.8 vs. 15.5–17.5 × 5–5.5 µm) and C. hystricis (10.5–12.5 × 3.3–4.8 vs. 13–15 × 4–5.5 µm).

Colletotrichum boninense Moriwaki, Toy. Sato & Tsukib., Mycoscience 44 (1): 48. 2003. (Figure 9).

Description: See Damm et al. [10].

Material examined: CHINA, Guizhou Province, Guiyang City, Aha Lake National Wetland Park, collected from diseased leaves of Hedera nepalensis, 106°36′59″ E, 26°33′55″ N, 27 June 2024, Weishan Zhang (HMAS 353991, living culture CFCC 72426); ibid. (living culture CFCC 72427); CHINA, Guizhou Province, Bijie City, Dafang County, collected from diseased leaves of Coriaria napalensis, 19 June 2024, 105°26′33″ E, 27°12′41″ N, Xinlei Fan (HMAS 353992, living culture CFCC 72424); ibid. (living culture CFCC 72425); CHINA, Guizhou Province, Bijie City, Dafang County, collected from diseased leaves of Fatsia japonica, 19 June 2024, 105°26′33″ E, 27°12′41″ N, Xinlei Fan (HMAS 353993, living culture CFCC 72422); ibid. (living culture CFCC 72423).

Culture characteristics: After culturing in the dark at 25 °C for 10 days, the colonies on PDA can reach 90 mm, white in color, with abundant flocculent aerial mycelium on the surface, spreading in appearance, and even margin. Conidial masses are produced after 15 days.

Notes: In the phylogenetic tree, the six isolates from this study clustered with C. boninense (Figure 3). Therefore, the six isolates were identified as C. boninense (Figure 2), representing one new host from China.

Colletotrichum crataegi W.S. Zhang & X.L. Fan, sp. nov. (Figure 10).

MycoBank: MB859293

Etymology: Named after the host genus of the type specimen, Crataegus.

Holotype: HMAS 353994

Description: Sexual morph not observed. Asexual morph: Sporulating on PDA. Conidiomata hyaline to light brown, oil droplets, acervulus, septate, unbranched, irregular. Conidiophores usually reduced to conidiogenous cells, unbranched. Conidiogenous cells (25.0–)29.0–46.5 × 2.5–5.7 μm (av. = 36.1 × 3.9 μm, n = 30), irregular, hyaline or light brown, smooth-walled, oil droplets. Conidia 9.5–13.5 × 3.0–3.8 μm (av. = 11.5 × 3.4 μm, n = 50), L/W ratio = 3.3, fusiform, hyaline, smooth-walled, contents granular. Appressoria circular to elliptical, light brown to dark brown, smooth-walled, 7.6–12.1 × 6.2–9.5(–11.2) μm (av. = 10.1 × 8.3 μm, n = 30). Setae not observed.

Culture characteristics: Colonies on PDA white, with even margin and abundant flocculent aerial mycelium, reaching 60 mm in diameter after 8 d at 25 °C. Black conidial masses are formed on the medium after 15 days.

Typus: CHINA, Beijing City, Mentougou District, Xiaolongmen Forest Farm, collected from diseased leaves of Crataegus pinnatifida, 22 August 2024, 115°25′00″ E, 39°48′34″ N, Xinlei Fan (holotype HMAS 353994; ex-holotype culture CFCC 72428); ibid. (living ex-paratype culture CFCC 72429).

Notes: Colletotrichum crataegi was associated with anthracnose of Crataegus pinnatifida. It clusters in a sister phylogenetic clade with C. lentis (BI/ML = 1.00/100) (Figure 4). However, the conidiogenous cells of C. crataegi are longer than those of C. lentis (29.0–46.4 vs. 9–28 µm). The conidia of C. crataegi are shorter than those of C. lentis (9.5–13.5 vs. 16–20 µm). The appressoria of C. crataegi are larger (7.6–12.1 × 6.2–9.5 vs. 5.5–7.5 × 4.5–6 µm). In addition, it differs from C. lentis in the nucleotide sequence by 9 bp in the ITS, 32 bp in gapdh, 14 bp in chs-1, 21 bp in act, 46 bp in tub2, and 34 bp in his3.

Colletotrichum dongguanense W.S. Zhang & X.L. Fan, sp. nov. (Figure 11).

MycoBank: MB859294

Etymology: Named after the collection location of the type specimen, Dongguan.

Holotype: HMAS 353998

Description: Sexual morph not observed. Asexual morph: Sporulating on PDA. Conidiomata and setae not observed. Conidiophores not developed. Conidiogenous cells formed from mycelium directly, branched. Conidia hyaline or pale brown, cylindrical or sub-cylindrical, aseptate, smooth-walled, no oil droplets, contents granular, 11.5–13.9(–16.2) × 4.5–6.5 μm (av. = 13.0 × 5.5 μm, n = 50), L/W ratio = 2.4. Appressoria single, both ends rounded or one end slightly pointed, medium to dark brown, 12.9–15.5(–23.9) × 5.8–7.7 μm (av. = 14.5 × 6.7 μm, n = 30).

Culture characteristics: Colonies on PDA 60 mm diam in 10 d, irregular margins, brown, with dense aerial mycelium, reverse black in the center, brown towards the margin. Black conidial masses are formed on the medium after 30 days.

Typus: CHINA, Guangdong Province, Dongguan City, Xingtang Xinghua Road, collected from diseased leaves of Bauhinia purpurea, 113°44′51″ E, 22°58′12″ N, 22 November 2023, Xinlei Fan (holotype HMAS 353998; ex-holotype culture CFCC 72438); ibid. (living culture CFCC 72439).

Notes: Colletotrichum dongguanense was associated with anthracnose of Bauhinia purpurea. Phylogenetically, C. dongguanense is sister to C. endophyticum and C. jinpingense (Figure 5). Colletotrichum dongguanense differs from C. endophyticum in the nucleotide sequence by 7 bp in gapdh, 2 bp in chs-1, 2 bp in act, and 3 bp in tub2. It also differs from C. jinpingense in nucleotide sequence by 4 bp in gapdh, 4 bp in chs-1, 3 bp in act, 4 bp in tub2, and 4 bp in cal. The mycelial color of C. endophyticum and C. jinpingense is gray on PDA medium, but C. dongguanense is brown. The conidial length of C. dongguanense is shorter than that of C. jinpingense (11.5–13.9 vs. 13.9–18.5 μm). The appressoria formed by C. dongguanense are regularly shaped, but those formed by C. endophyticum and C. jinpingense are irregularly shaped. Moreover, the appressoria of C. dongguanense are longer than those of C. endophyticum (12.9–15.5 vs. 8–12 μm).

Colletotrichum fioriniae (Marcelino & Gouli) Pennycook., Mycotaxon 132 (1): 150. 2017.

Description: See Zhang et al. [7].

Material examined: CHINA, Shaanxi Province, Ankang City, Hanbin District, Zigou Town, Erlang Village, collected from diseased leaves of Juglans regia, 108°57′32″ E, 32°55′24″ N, 14 May 2024, Lu Lin & Xinlei Fan (living culture CFCC 72578 and CFCC 72579); CHINA, Guizhou Province, Bijie City, Dafang County, Jinhai Lake Wetland Park, collected from diseased leaves of Ulmus parvifolia, 105°26′30″ E, 27°12′45″ N, 19 June 2024 Xinlei Fan (living culture LFPR 10003 and LFPR 10005); CHINA, Fujian Province, Fuzhou City, Jinan District, Fengchi Baiyun Cave Scenic Area, collected from diseased leaves of Laurus nobilis, 119°17′27″ E, 26°6′9″ N, 19 August 2024 Xinlei Fan (living culture LFPR 10004).

Notes: In the phylogenetic tree, the five isolates from this study clustered with C. fioriniae (BI/ML = 1.00/100) (Figure 2). Therefore, the five isolates were identified as C. fioriniae (Figure 2), representing three new hosts from China.

Colletotrichum flavosporum W.S. Zhang & X.L. Fan, sp. nov. (Figure 12).

MycoBank: MB859297

Etymology: Named after the rare characteristic of this new species, flavosporum.

Holotype: HMAS 353997

Description: Sexual morph not observed. Asexual morph: Sporulating on PDA. Conidiomata scattered or gregarious, semi-immersed to immersed, hyaline. Conidiophores usually reduced to conidiogenous cells, branched. Conidiogenous cells hyaline or pale brown, straight or curved, cylindrical or ampulliform, smooth-walled, 19.2–22.8(–33.5) × 2.1–4.3 μm (av. = 21.1 × 3.4 μm, n = 50). Conidia cylindrical or sub-cylindrical, hyaline, contents granular, aseptate, smooth-walled, no oil droplets, 10.5–11.9 × 4.5–5.4 μm (av. = 11.2 × 5.0 μm, n = 50), L/W ratio = 2.2. Appressoria single, circular to irregular, hyaline or dark brown, (13.8–)17.1–20.5 × 7.5–10.5 μm (av. = 18.7 × 9.1 μm, n = 30). Setae not observed.

Culture characteristics: Colonies on PDA reach a diameter of 50 mm after 7 d at 25 °C, with regular margins, white in color, flat, and featuring dense aerial mycelium that exhibits a spreading growth habit.

Typus: CHINA, Guangdong Province, Dongguan City, Xingtang Xinghua Road, collected from diseased leaves of Bougainvillea glabra, 113°44′53″ E, 22°57′38″ N, 19 December 2024, Xinlei Fan (holotype HMAS 353997; ex-holotype culture CFCC 72442); ibid. (living culture CFCC 72443).

Notes: Colletotrichum flavosporum was associated with anthracnose of Bougainvillea glabra. Phylogenetically, C. flavosporum is closely related to C. dracaenigenum (Figure 5), but it differs from C. dracaenigenum in nucleotide sequence by 6 bp in ITS and 8 bp in gapdh. Morphologically, C. flavosporum produces branched conidiophores. The conidiogenous cells of C. flavosporum are longer than those of C. dracaenigenum (19.2–22.8 vs. 13–15 μm). The appressoria of C. flavosporum are larger than those of C. dracaenigenum (17.1–20.5 × 7.5–10.5 vs. 5–12 × 4–7 μm).

Colletotrichum fructicola Prihast. et al., Fungal Diversity 39: 158. 2009.

Description: See de Silva et al. [22].

Material examined: CHINA, Beijing City, Shunyi District, County Road 408, collected from diseased leaves of Juglans regia, 116°39′17″ E, 40°7′49″ N, 10 October 2024, Xinlei Fan (living culture CFCC 72434 and CFCC 72435).

Notes: In this study, the two strains clustered with C. fructicola in a subclade, with 94% ML value and 1.00 BI value (Figure 5). The ITS, chs-1, act, tub2, and cal genes of the two strains were identical to those of C. fructicola, with only a 2 bp difference in gapdh (309/311, 99.4%), and they were morphologically similar. Therefore, we identified them as C. fructicola.

Colletotrichum gloeosporioides (Penz.) Penz. & Sacc., Atti del Reale Istituto Veneto di Scienze, Lettere ed Arti, 6, 2 (5): 670. 1884. (Figure 13).

New proposed synonyms.

Colletotrichum peakense L. Zhang et al. Mycokeys 99: 141 (2023).

Colletotrichum juglandicola L. Zhang et al. Mycokeys 99: 139 (2023).

Colletotrichum juglandium Y.X. Li et al. Mycokeys 108: 158 (2024).

Description: Sexual morph not observed. Asexual morph: Sporulating on PDA. Conidiomata scattered or gregarious, semi-immersed, hyaline. Conidiophores usually reduced to conidiogenous cells, unbranched, aseptate. Conidiogenous cells hyaline, straight, cylindrical, smooth-walled, 23.5–33.5 × 1.7–2.8 μm (av. = 27.3 × 2.2 μm, n = 50). Conidia cylindrical, hyaline, contents granular, smooth-walled, no oil droplets, 14.9–17.5 × 4.0–5.2 μm (av. = 16.1 × 4.6 μm, n = 50), L/W ratio = 3.4. Setae medium to dark brown, single, circular to irregular, 3–4-septate, 36–43 μm. Appressoria not observed.

Material examined: CHINA, Beijing City, Shunyi District, County Road 408, collected from diseased leaves of Juglans regia, 116°39′15″ E, 40°7′52″ N, 10 July 2024, Xinlei Fan (living culture CFCC 72580). ibid. 11 October 2024, Xinlei Fan (living culture CFCC 72581). ibid. 116°39′15″ E, 40°7′49″ N, 10 July 2024, Xinlei Fan (living culture LFPR 10014); CHINA, Beijing City, Mentougou District, Xiaolongmen Forest Farm, collected from diseased leaves of Juglans regia, 115°25′13″ E, 39°48′45″ N, 22 August 2024, Xinlei Fan (living culture LFPR 10025). CHINA, Beijing City, Haidian District, Cuihu National Urban Wetland Park, collected from diseased leaves of Prunus cerasifera ‘Atropurpurea’, 116°39′17″ E, 40°7′49″ N, 18 July 2024, Weishan Zhang (living culture CFCC 72440 and CFCC 72441). ibid. collected from diseased leaves of Robinia pseudoacacia, 116°10′45″ E, 40°6′10″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10019); ibid. collected from diseased leaves of Fraxinus chinensis, 116°10′58″ E, 40°6′10″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10020); ibid. collected from diseased leaves of Prunus persica ‘Duplex’, 116°11′21″ E, 40°6′18″ N, 13 October 2024, Weishan Zhang (living culture LFPR 10022); ibid. collected from diseased leaves of Malus spectabilis, 116°11′15″ E, 40°6′19″ N, 13 October 2024, Weishan Zhang (living culture LFPR 10023); ibid. collected from diseased leaves of Kerria japonica, 116°11′17″ E, 40°6′21″ N, 13 October 2024, Weishan Zhang (living culture LFPR 10024); ibid. collected from diseased leaves of Amorpha fruticosa, 116°11′6″ E, 40°6′9″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10012); ibid. 116°11′9″ E, 40°6′9″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10013); ibid. 116°11′15″ E, 40°6′20″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10021). CHINA, Guizhou Province, Guiyang City, Qianling Mountain Park, collected from diseased leaves of Quercus aliena, 106°41′44″ E, 26°35′27″ N, 19 June 2024, Xinlei Fan (living culture LFPR 10015); ibid. Aha Lake National Wetland Park, collected from diseased leaves of Parthenocissus quinquefolia, 106°36′59″ E, 26°33′57″ N, 27 June 2024, Xinlei Fan (living culture LFPR 10016); CHINA, Shanxi Province, Ankang City, Hanbin District, Zigou Town, Erlang Village, collected from diseased leaves of Parthenocissus tricuspidata, 108°57′52″ E, 32°55′40″ N, 13 June 2024, Xinlei Fan (living culture LFPR 10017); ibid. collected from diseased leaves of Microlepia marginata, 108°57′50″ E, 32°55′41″ N, 13 June 2024 Xinlei Fan (living culture LFPR 10018).

Notes: Phylogenetically, the four strains are closely related to C. gloeosporioides, C. juglandium, C. juglandicola, and C. peakense. (BI/ML = 1.00/100) (Figure 5). Strains CFCC 72580 and CFCC 72581 had no base differences in the ITS, gapdh, chs-1, act, tub2, and cal when compared with C. gloeosporioides and C. peakense. Strains CFCC 72440 and CFCC 72441 had base differences in gapdh (10 bp) compared with C. gloeosporioides. The isolates in this study had base differences with C. juglandicola (ITS: 1 bp; chs-1: 1 bp; tub2: 1 bp). Our strains had base differences in gapdh (3 bp) compared with C. juglandium. Our strains were morphologically similar to C. gloeosporioides. Therefore, we identified all four strains as C. gloeosporioides. In addition, based on the phylogenetic tree and the lack of sequence variation, we regarded C. juglandium, C. juglandicola, and C. peakense as synonyms of C. gloeosporioides. A detailed explanation for the taxonomic treatment of this section is provided in the discussion.

Colletotrichum godetiae Neerg., Friesia 4 (1–2): 72. 1950.

New proposed synonyms.

Colletotrichum americanum M. Zapata et al. Mycological Progress 23: 28. 2024.

Description: See Damm et al. [9].

Material examined: CHINA, Shanxi Province, Ankang City, Hanbin District, Cigou Town, Er Lang Village, collected from diseased leaves of Juglans regia, 108°57′57″ E, 32°55′42″ N, 14 June 2024, Xinlei Fan (living culture CFCC 72574 and CFCC 72575). ibid. 108°57′52″ E, 32°55′40″ N, 14 June 2024, Xinlei Fan (living culture LFPR 10001). CHINA, Guizhou Province, Bijie City, Dafang County, Jinhai Lake Wetland Park, collected from diseased leaves of Calystegia hederacea, 105°26′31″ E, 27°12′44″ N, 19 June 2024 Weishan Zhang (living culture LFPR 10002).

Notes: In the phylogenetic tree, the two strains in this study formed a closely related clade with C. americanum and C. godetiae (BI/ML = 0.99/95) (Figure 2). Our two strains had base differences with C. godetiae (ITS: 1 bp; gapdh: 1 bp). However, our isolates had no base differences in other gene fragments with C. godetiae. Therefore, we identified the two strains in this study as C. godetiae. C. americanum (RGM 3380T and RGM 3407) and C. godetiae CBS 862.70 had no base differences and morphologically similar. Based on this, we regarded C. americanum as a synonym of C. godetiae. A detailed explanation for the taxonomic treatment of this section is provided in the discussion.

Colletotrichum karsti Y.L. Yang, Z.Y. Liu, K.D. Hyde & L. Cai, Cryptogonia Mycologia 32 (3): 241. 2011.

Description: See Zhang et al. [7].

Material examined: CHINA, Fujian Province, Fuzhou City, Jinan District, National Wetland Park, collected from diseased leaves of Acer rubrum, 119°19′25″ E, 26°5′6″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10008). CHINA, Guizhou Province, Guiyang City, Aha Lake National Wetland Park, collected from diseased leaves of Parthenocissus quinquefolia, 106°36′53″ E, 26°33′52″ N, 27 June 2024, Weishan Zhang (living culture LFPR 10009).

Notes: In the phylogenetic tree, the two strains in this study clustered with C. karsti (BI/ML = 1.00/100) (Figure 3). Therefore, these two isolates were identified as C. karsti (Figure 3).

Colletotrichum nymphaeae (Pass.) Aa., Netherlands Journal of Plant Pathology 84 (3): 110. 1978.

Description: See Damm et al. [9].

Material examined: CHINA, Shanxi Province, Ankang City, Hanbin District, Cigou Town, Er Lang Village, collected from diseased leaves of Juglans regia, 108°57′21″ E, 32°55′18″ N, 14 May 2024, Xinlei Fan (living culture CFCC 72576 and CFCC 72577). CHINA, Guizhou Province, Bijie City, Dafang County, Jinhai Lake Wetland Park, collected from diseased leaves of Senecio scandens, 105°26′37″ E, 27°12′45″ N, 19 June 2024 Weishan Zhang (living culture LFPR 10007). CHINA, Guizhou Province, Guiyang City, Aha Lake National Wetland Park, collected from diseased leaves of Fatsia japonica, 106°36′51″ E, 26°33′59″ N, 27 June 2024, Weishan Zhang (living culture LFPR 10006).

Notes: In the phylogenetic tree, our two isolates (CFCC 72576 and CFCC 72577) clustered with C. nymphaeae (BI/ML = 1.00/98) (Figure 2). Therefore, these two isolates were identified as C. nymphaeae (Figure 2), representing a new host from China.

Colletotrichum orchidearum Allesch., Rabenh. Krypt.-Fl., Edn 2 (Leipzig) 1 (7): 563. 1903.

Description: See Damm et al. [38].

Material examined: CHINA, Fujian Province, Fuzhou City, Jinan District, National Wetland Park, collected from diseased leaves of Anthurium andraeanum, 119°19′25″ E, 26°5′6″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10029); ibid. 119°19′21″ E, 26°5′9″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10030).

Notes: In the phylogenetic tree, our two isolates (LFPR 10029 and LFPR 10030) clustered with C. orchidearum (BI/ML = 1.00/100) (Figure 6). Therefore, these two isolates were identified as C. orchidearum (Figure 6), representing a novel geographic record for China.

Colletotrichum plurivorum U. Damm., Alizadeh & T. Sato., Studies in Mycology 92: 31. 2018.

New proposed synonyms. Colletotrichum subplurivorum Sui et al. Mycosphere 15 (1): 4569–4743. 2024.

Description: See Zhang et al. [7].

Material examined: CHINA, Fujian Province, Fuzhou City, Jinan District, National Wetland Park, collected from diseased leaves of Nandina domestica, 119°19′27″ E, 26°5′6″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10026); ibid. collected from diseased leaves of Spathiphyllum, 119°19′25″ E, 26°5′8″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10027); ibid. collected from diseased leaves of Megaskepasma erythrochlamys, 119°19′22″ E, 26°5′6″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10028).

Notes: In the phylogenetic tree, the three strains in this study formed a closely related clade with C. plurivorum and C. subplurivorum (BI/ML = 1.00/98) (Figure 6). Our strain (LFPR 10028) had base differences with C. plurivorum (LC8337) (ITS: 1 bp; tub2: 5 bp). However, our isolates had no base differences in other gene fragments with C. plurivorum (LC8337). Therefore, we identified the three strains in this study as C. plurivorum. C. subplurivorum (CNUCC 833B-1-1 T) had base differences with C. plurivorum (LC8337) (ITS: 5 bp; gapdh:1 bp; tub2: 1 bp). Based on the lack of sequence variation, we regarded C. subplurivorum as a synonym of C. plurivorum.

Colletotrichum siamense Prihast., L. Cai & K.D. Hyde., Fungal Diversity 39: 98. 2009. (Figure 14).

Description: See Zhang et al. [7].

Material examined: CHINA, Fujian Province, Fuzhou City, Jinan District, National Wetland Park, collected from diseased leaves of Nandina domestica, 119°19′27″ E, 26°5′9″ N, 20 August 2024, Weishan Zhang (living culture LFPR 10011); CHINA, Beijing City, Haidian District, Cuihu National Urban Wetland Park, collected from diseased leaves of Broussonetia papyrifera, 116°11′60″ E, 40°5′24″ N, 13 October 2024, Weishan Zhang (living culture LFPR 10010). CHINA, Guangdong Province, Dongguan City, Xingtang Xinghua Road, collected from diseased leaves of Lagerstroemia speciosa, 113°44′38″ E, 22°59′33″ N, 14 November 2023, Xinlei Fan (HMAS 353995; living culture CFCC 72442); ibid. (living culture CFCC 72443). CHINA, Beijing City, Fengtai District, Lotus Pool Park, collected from diseased branch of Euonymus japonicus, 2 July 2024, 116°18′49″ E, 39°53′27″ N, Xinlei Fan (BJFC-S2404; living culture CFCC 72605); ibid. (BJFC-S2405; living culture CFCC 72606). CHINA, Fujian Province, Fuzhou City, Jinan District, National Wetland Park, collected from diseased leaves of Chamaedorea pinnatifrons, 20 August 2024, 119°19′25″ E, 26°5′6″ N, Weishan Zhang (HMAS 353999; living culture CFCC 72430); ibid. (living culture CFCC 72431).

Notes: In the phylogenetic tree, the eight strains in this study clustered with C. siamense (Figure 5). The eight strains exhibit morphological similarity to C. siamense. Therefore, these eight isolates were identified as C. siamense.

Colletotrichum sojae Damm., & Alizadeh., Studies in Mycology, 92: 35. 2018.

Description: See Sui et al. [2].

Material examined: CHINA, Beijing City, Haidian District, Cuihu National Urban Wetland Park, collected from diseased leaves of Lonicera maackii, 116°11′59″ E, 40°5′26″ N, 13 October 2024, Weishan Zhang (living culture LFPR 10029); ibid. 116°11′60″ E, 40°5′26″ N, 13 October 2024, Weishan Zhang (living culture LFPR 10030).

Notes: In the phylogenetic tree, the two strains in this study clustered with C. sojae (BI/ML = 1.00/100) (Figure 6). Therefore, these two isolates were identified as C. sojae.

Colletotrichum spaethianum (Allesch.) Damm., P.F. Cannon & Crous, Fungal Diversity. 39: 74. 2009.

Description: See Damm et al. [39].

Material examined: CHINA, Beijing City, Haidian District, Cuihu National Urban Wetland Park, collected from diseased leaves of Hosta plantaginea, 116°11′51″ E, 40°6′5″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10033); ibid. collected from diseased leaves of Malus pumila, 116°11′50″ E, 40°6′5″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10034); ibid. 116°11′50″ E, 40°6′6″ N, 18 July 2024, Weishan Zhang (living culture LFPR 10035).

Notes: In the phylogenetic tree, the three strains in this study clustered with C. spaethianum (BI/ML = 1.00/100) (Figure 7). Therefore, these three isolates were identified as C. spaethianum.

3.3. Correlation Analysis Between Pathogens and Hosts

The 67 strains identified in this study belong to 16 Colletotrichum species. The species in Beijing are the most diverse (including six Colletotrichum species, one of which is a new Colletotrichum species) (Figure 15a). The other three new Colletotrichum species are all from Guangdong, possibly due to geographic barriers limiting their spread (Figure 15a). C. gloeosporioides has the highest isolation rate and a wide distribution (Figure 15a). C. gloeosporioides is concentrated in Beijing, Shaanxi, and Guizhou (Figure 15a). C. gloeosporioides has a diverse range of hosts, including Amorpha fruticosa, Juglans regia, Parthenocissus quinquefolia, Quercus aliena var. acuteserrata, and Prunus cerasifera ‘Atropurpurea’, showing its broad-host characteristics (Figure 15b). Walnut is dominant among the hosts of C. gloeosporioides, accounting for 22%, which may be related to its leaf morphology or weaker disease resistance (Figure 15b).

4. Discussion

This study has identified 16 Colletotrichum species associated with anthracnose diseases in Chinese plants, including three known species within the C. acutatum complex, four known species within the C. boninense complex, one new species within the C. destructivum complex, six new species and three known species within the C. gloeosporioides complex, three known species within the C. orchidearum, and one known species within the C. spaethianum. These findings highlight the remarkable diversity of Colletotrichum species causing anthracnose diseases in China.

Jayawardena et al. [40] suggested that Colletotrichum species have complex ecological adaptability. The C. acutatum, C. boninense, and C. gloeosporioides complexes exhibit greater species diversity and broader host ranges compared to others [16]. This study also includes ten new host records, such as Hedera nepalensis for C. boninense, Aquilaria sinensis for C. aquilariae, Bauhinia purpurea for C. dongguanense, Bougainvillea glabra for C. flavosporum, and Juglans regia for both C. fioriniae and C. nymphaeae. The broad host range of these species may be associated with evolutionary processes, ecological niche differentiation, and host interactions.

The research results reveal the ecological characteristics of Colletotrichum species from the dimensions of species diversity, geographical distribution, and host preference. In terms of species diversity and geographical differentiation, Beijing has the highest species richness of Colletotrichum (including 6 species, one of which is a new species), reflecting that habitat heterogeneity provides a niche basis for fungal differentiation; Guangdong has isolated 3 new species. Combined with the speculation that “geographical barriers limit dispersal”, this confirms that geographical isolation drives the formation of new species—isolated environments reduce gene flow and promote population specialization.

Colletotrichum gloeosporioides exhibits prominent ecological dominance: this species has the highest isolation rate and is widely distributed in Beijing, Shaanxi, and Guizhou, reflecting its strong adaptability to different climatic and soil conditions; from the host dimension, its hosts cover 11 plant families, including Amorpha fruticosa, Juglans regia, Parthenocissus quinquefolia, etc., demonstrating broad–spectrum parasitism. However, walnut accounts for 22% of the hosts (significantly higher than other plants). Combined with the speculation that “leaf morphology or weak disease resistance” may be involved, it suggests that the leaf microstructure of walnut (such as stomatal density and cuticle thickness) or defects in its own defense pathways are easily exploited by the pathogen, which provides a core target for walnut disease control. Walnut (Juglans regia L.), as a key host, is particularly associated with multiple Colletotrichum species. This reveals the susceptibility of walnut due to physiological and environmental factors and the consistency with the strains reported by Li et al. [32]. It further emphasizes the stability and universality of these host–pathogen interactions.

Taxonomic uncertainties were observed within the C. acutatum, C. gloeosporioides, and C. orchidearum species complexes. The strains in this study within the C. acutatum complex showed close relationships with C. americanum and C. godetiae. Although Zapata et al. [41] introduced C. americanum, they did not conduct phylogenetic or morphological comparisons with the C. godetiae CBS 862.70 strain. Here, we demonstrate that C. americanum (RGM 3380T and RGM 3407) and C. godetiae CBS 862.70 are morphologically similar and exhibit no significant genetic differences. Therefore, we propose that C. americanum is a synonym of C. godetiae. The current strains formed a unique clade with C. gloeosporioides, C. juglandicola, C. juglandium, and C. peakense within the C. gloeosporioides species complex. Previous studies have shown conflicting relationships among these species: Zhang et al. [21] reported that C. juglandium and C. peakense are closely related to C. dimorphum, while Zhang et al. [31] suggested that C. dimorphum is a synonym of C. gloeosporioides. The current expanded analysis, incorporating additional strains of C. gloeosporioides, C. juglandicola, C. juglandium, and C. peakense, revealed that they consistently clustered into a stable clade (Figure 5). These three species exhibited less than 1% sequence divergence and shared overlapping morphological features. Based on these findings, we propose that C. juglandicola, C. juglandium, and C. peakense should be considered synonyms of C. gloeosporioides. The strains in this study within the C. orchidearum complex showed close relationships with C. subplurivorum and C. orchidearum. Sui et al. [18] introduced C. subplurivorum, but they did not conduct a phylogenetic comparison with C. orchidearum (LC8337). In this study, we demonstrate that C. subplurivorum (CNUCC 833B-1-1T) and C. orchidearum (LC8337) exhibit no significant genetic differences. Therefore, we assume that C. subplurivorum might be a synonym of C. orchidearum.

The identification of Colletotrichum species mostly relies on multigene phylogenetic analysis combined with morphological identification. With the development of molecular techniques, the application of high-throughput sequencing technology has also provided new means for the identification of Colletotrichum species. For example, Liu et al. (2022) [16] conducted high-throughput sequencing of Colletotrichum samples, generating and assembling whole-genome sequences for 30 new species and 18 known species. This demonstrates the great potential of high-throughput sequencing technology in fungal taxonomy. Moreover, although these species were isolated from diseased samples, their exact pathogenic mechanisms remain unclear. Therefore, further research is crucial for elucidating the pathogenic mechanisms of these pathogens.