Min Qiao

Colletotrichum spp. are widespread fungal pathogens that cause anthracnose in numerous economically important crops and, exhibiting extensive taxonomic, host plant, and lifestyle diversity. Here, we analyzed the genome sequences of 150 strains representing 97 species across 15 species complexes and four singletons, including and integrating both newly assembled and publicly available genomes. Phylogenomic investigation clarified the taxonomy of Colletotrichum and resolved misidentifications. We identified variations in genome architecture contributed by phylogenetic lineages, host types, and lifestyles, with transposable element proliferation playing significant roles. Interestingly, codon usage bias followed phylogenetic patterns, with species complexes forming distinct clusters and exhibiting a significant bistable co-evolutionary relationship with tRNA genes. Functional gene repertoires displayed coordinated shifts, with higher abundance in broad host-range species complexes and in strains associated with woody or dicotyledonous hosts. Although most functional categories retained strong phylogenetic signals, co-occurrence analysis of weak-signal categories identified modules related to host cell wall disruption, fungal cell wall remodeling, and virulence that were significantly associated with ecological differentiation. Evolutionary trajectories and gene family dynamics further revealed divergent ecological strategies, with oxidative versus rapid-response detoxification in woody- and herbaceous-associated lineages, respectively. The diversifications were accompanied by woody-specific expansion of GH39 and alkaline proteases and progressive differentiation of pectin-degrading capacity, including contraction in woody lineages and divergence between dicot- and monocot-associated herbaceous lineages. The C. gloeosporioides species complex emerged with a comprehensive expansion of detoxification and cell wall-degrading capacities, likely contributing to its broad host range. In contrast, endophytic lineages exhibited convergent gene family contraction in adhesion and cell wall remodeling. Together, this study revealed concordance of codon usage and functional gene abundance with phylogeny, along with diverse host- and lifestyle-associated adaptive strategies in this important group of plant pathogens.

Introduction: Trichoderma species establish symbiotic relationships with plants through both parasitic and mutualistic mechanisms. While some Trichoderma species act as plant pathogenic fungi, others utilize various strategies to protect and enhance plant growth.Methods: Phylogenetic positions of new species of Trichoderma were determined through multi-gene analysis relying on the internal transcribed spacer (ITS) regions of the ribosomal DNA, the translation elongation factor 1-α (tef1-α) gene, and the RNA polymerase II (rpb2) gene. Additionally, pathogenicity experiments were conducted, and the aggressiveness of each isolate was evaluated based on the area of the cross-section of the infected site.Results: In this study, 13 Trichoderma species, including 9 known species and 4 new species, namely, T. delicatum, T. robustum, T. perfasciculatum, and T. subulatum were isolated from the diseased tubers of Gastrodia elata in Yunnan, China. Among the known species, T. hamatum had the highest frequency. T. delicatum belonged to the Koningii clade. T. robustum and T. perfasciculatum were assigned to the Virens clade. T. subulatum emerged as a new member of the Spirale clade. Pathogenicity experiments were conducted on the new species T. robustum, T. delicatum, and T. perfasciculatum, as well as the known species T. hamatum, T. atroviride, and T. harzianum. The infective abilities of different Trichoderma species on G. elata varied, indicating that Trichoderma was a pathogenic fungus causing black rot disease in G. elata.Discussion: This study provided the morphological characteristics of new species and discussed the morphological differences with phylogenetically proximate species, laying the foundation for research aimed at preventing and managing diseases that affect G. elata.

Black rot is a common disease of Gastrodia elata, causing serious threats to G. elata production. In this study, a total of 17 Cylindrocarpon-like strains were isolated from G. elata black rot tissues. Multilocus sequence analyses based on ITS, HIS, TEF, and TUB combined with morphological characterizations were performed to identify six Ilyonectria species, including four new species, Ilyonectria longispora, I. sinensis, I. xiaocaobaensis, and I. yunnanensis, and two known species, I. changbaiensis and I. robusta. The pathogenicity of 11 isolates comprising type strains of the four new species and representative isolates from each of the six species was tested on healthy tissues of G. elata. All isolates were pathogenic to G. elata tissues, and symptoms were identical to black rot disease, confirming that our isolates were the causal agents of black rot disease of G. elata.

Background: Aspergillus niger is generally considered safe and is widely used across a broad range of industries from food to pharmaceuticals. The metabolites of some Aspergillus species are effective in killing nematodes. The active ingredients that play a role are citric acid and oxalic acid.Results: The effective nematocidal metabolite in the fermentation broth of the A. niger Ym16 is a calcium oxalate coordination compound (C₂H₂O₄•2(C₂HO₄)•C₂O₄•4(H₂O)•2(Ca)). The mortality of fermentation broth and compound solution to second‐stage juveniles (J2s) of Meloidogyne incognita were 94% (8 h) and 95% (2 h), respectively. The hatching inhibition rates of fermentation broth and compound solution to eggs of M. incognita were 86% (24 h) and 87% (12 h), respectively. Furthermore, the Ym16 strain promoted plant growth.Conclusion: Strain Ym16 and its metabolites should be the first choice for nematode control because it is an effective agent against soil nematodes. © 2023 Society of Chemical Industry.

During a survey of soil fungal diversity in a rocky desertification area in Yunnan Province, China, a new dematiaceous hyphomycete, Cordana yunnanensis was identified. Morphologically, this species is characterized by macronematous, mononematous conidiophores with discrete, polyblastic conidiogenous cells arranged in a whorl at the apices. Phylogenetic analysis of the combined sequences of the internal transcribed spacer and the large nuclear ribosomal RNA subunit confirmed the phylogenetic position of C. yunnanensis within the genus Cordana, in Cordanaceae.

An unknown endophytic fungus was isolated from the aquatic plant Hippuris vulgaris in Litang county, Sichuan province, China. Phylogenetic analyses inferred from combined ITS, Sod2, Apn2, and TUB2 sequences revealed that the endophyte is a new species belonging to the Colletotrichum graminicola species complex. Morphological characteristics showed that Colletotrichum litangense is characterized by its falcate, lunate to sublunate conidia, and ellipsoidal, ovoid, or lobed appressoria. Pathogenicity tests on several fruits showed that C. litangense could induce anthracnose lesions. As a result of the phylogenetic, morphological, and pathogenicity analyses, we proposed the name Colletotrichum litangense for the new species.

Ageratina adenophora is one of the most invasive weeds in China. Following an outbreak in Yunnan in the 1960s, A. adenophora has been spreading in Southwest China at tremendous speed. Previous research indicated A. adenophora contained many Colletotrichum species as endophytes. In this study, we investigated the diversity of Colletotrichum in healthy and diseased leaves of the invasive plant A. adenophora and several surrounding crops in Yunnan, Guangxi, and Guizhou provinces in China, and obtained over 1000 Colletotrichum strains. After preliminary delimitation using the internal transcribed spacer region (ITS) sequences, 44 representative strains were selected for further study. Their phylogenetic positions were determined by phylogenetic analyses using combined sequences of ITS, actin (ACT), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-tubulin (TUB2). Combined with morphological characteristics, 14 new Colletotrichum species were named as C. adenophorae, C. analogum, C. cangyuanense, C. dimorphum, C. gracile, C. nanhuaense, C. nullisetosum, C. oblongisporum, C. parvisporum, C. robustum, C. simulanticitri, C. speciosum, C. subhenanense, and C. yunajiangense.

Colletotrichum species are plant pathogens, saprobes, and endophytes in many economically important hosts. Many studies have investigated the diversity and pathogenicity of Colletotrichum species in common ornamentals, fruits, and vegetables. However, Colletotrichum species occurring in aquatic plants are not well known. During the investigation of the diversity of endophytic fungi in aquatic plants in southwest China, 66 Colletotrichum isolates were obtained from aquatic plants there, and 26 of them were selected for sequencing and analyses of actin (ACT), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the internal transcribed spacer (ITS) region, and β-tubulin (TUB2) genomic regions. Based on morphological characterization and multi-locus phylogenetic analyses, 13 Colletotrichum species were recognized, namely, C. baiyuense sp. nov., C. casaense sp. nov., C. demersi sp. nov., C. dianense sp. nov., C. fructicola, C. garzense sp. nov., C. jiangxiense, C. karstii, C. philoxeroidis sp. nov., C. spicati sp. nov., C. tengchongense sp. nov., C. vulgaris sp. nov., C. wuxuhaiense sp. nov. Two species complexes, the C. boninense species complex and C. gloeosporioides species complex, were found to be associated with aquatic plants. Pathogenicity tests revealed a broad diversity in pathogenicity and aggressiveness among the eight new Colletotrichum species.

A new species, Colletotrichum menglaense, isolated from air in Mengla, Xishuangbanna, Yunnan Province, China, was characterized and described combining morphological characteristics and multigene phylogenetic analysis. Morphologically, it is characterized by oblong, sometimes slightly constricted, micro-guttulate conidia and simple obovoid to ellipsoidal appressoria. Phylogenetic analysis of the ITS, ACT, CHS, and GAPDH sequences showed that C. menglaense belongs to the C. gloeosporioides complex. The pathogenicity of C. menglaense on fruits of several crop plants, including strawberry, orange, grape, tomato, and blueberry, was tested and confirmed by the re-isolation of C. menglaense.

Aquatic ecosystems contain tremendous plant and microbial diversity. However, little is known about endophyte diversity in aquatic plants. In this study, we investigated the diversity of endophytic fungi in aquatic plants in southwest China using both culture-based and culture-independent high-throughput sequencing methods. A total of 1,689 fungal isolates belonging to three phyla and 154 genera were obtained from 15,373 plant tissue segments of 30 aquatic plant species. The most abundant endophytic fungi were those in ascomycete genera Aspergillus, Ceratophoma, Fusarium, Penicillium, Phoma and Plectosporium. No difference in fungal isolation rates was observed among tissues from roots, stems, and leaves. Twenty tissue samples from three most common plant species were further subjected to culture-independent meta-barcode sequencing. The sequence-based analyses revealed a total of 1,074 OTUs belonging to six fungal phyla and 194 genera. Among the three plants, Batrachium bungei harbored the highest number of OTUs. Besides, a total of 66 genera were detected by two methods. Both the culture-dependent and independent methods revealed that aquatic plants in southwest China have abundant endophytic fungal diversity. This study significantly expands our knowledge of the fungal community of aquatic plants.