Novel species of fungi described in this study include those from various countries as follows: Argentina, Colletotrichum araujiae on leaves, stems and fruits of Araujia hortorum. Australia, Agaricus pateritonsus on soil, Curvularia fraserae on dying leaf of Bothriochloa insculpta, Curvularia millisiae from yellowing leaf tips of Cyperus aromaticus, Marasmius brunneolorobustus on well-rotted wood, Nigrospora cooperae from necrotic leaf of Heteropogon contortus, Penicillium tealii from the body of a dead spider, Pseudocercospora robertsiorum from leaf spots of Senna tora, Talaromyces atkinsoniae from gills of Marasmius crinis-equi and Zasmidium pearceae from leaf spots of Smilax glyciphylla. Brazil, Preussia bezerrensis from air. Chile, Paraconiothyrium kelleni from the rhizosphere of Fragaria chiloensis subsp. chiloensis f. chiloensis. Finland, Inocybe udicola on soil in mixed forest with Betula pendula, Populus tremula, Picea abies and Alnus incana. France, Myrmecridium normannianum on dead culm of unidentified Poaceae. Germany, Vexillomyces fraxinicola from symptomless stem wood of Fraxinus excelsior. India, Diaporthe limoniae on infected fruit of Limonia acidissima, Didymella naikii on leaves of Cajanus cajan, and Fulvifomes mangroviensis on basal trunk of Aegiceras corniculatum. Indonesia, Penicillium ezekielii from Zea mays kernels. Namibia, Neocamarosporium calicoremae and Neocladosporium calicoremae on stems of Calicorema capitata, and Pleiochaeta adenolobi on symptomatic leaves of Adenolobus pechuelii. Netherlands, Chalara pteridii on stems of Pteridium aquilinum, Neomackenziella juncicola (incl. Neomackenziella gen. nov.) and Sporidesmiella junci from dead culms of Juncus effusus. Pakistan, Inocybe longistipitata on soil in a Quercus forest. Poland, Phytophthora viadrina from rhizosphere soil of Quercus robur, and Septoria krystynae on leaf spots of Viscum album. Portugal (Azores), Acrogenospora stellata on dead wood or bark. South Africa, Phyllactinia greyiae on leaves of Greyia sutherlandii and Punctelia anae on bark of Vachellia karroo. Spain, Anteaglonium lusitanicum on decaying wood of Prunus lusitanica subsp. lusitanica, Hawksworthiomyces riparius from fluvial sediments, Lophiostoma carabassense endophytic in roots of Limbarda crithmoides, and Tuber mohedanoi from calcareus soils. Spain (Canary Islands), Mycena laurisilvae on stumps and woody debris. Sweden, Elaphomyces geminus from soil under Quercus robur. Thailand, Lactifluus chiangraiensis on soil under Pinus merkusii, Lactifluus nakhonphanomensis and Xerocomus sisongkhramensis on soil under Dipterocarpus trees. Ukraine, Valsonectria robiniae on dead twigs of Robinia hispida. USA, Spiralomyces americanus (incl. Spiralomyces gen. nov.) from office air. Morphological and culture characteristics are supported by DNA barcodes.
We describe the first identification of Gnomoniopsis smithogilvyi causing brown rot on chestnut fruits in Chile, with an incidence of 4.8%. Previously, Phomopsis castanea (IMI 278057) was reported as the cause of the disease in Chile, but a molecular re-identification revealed that it corresponded to G. smithogilvyi. All chestnut fruits inoculated with the isolate G. smithogilvyi RGM 2903 developed brown rot symptoms on fruits.
Vaccinium corymbosum L. is the most cultivated blueberry species in Chile. Chilean fruits typically take up to 50 days to reach oversea markets; therefore, controlling post-harvest pathogens is of outmost importance to maintain international food safety and quality standards. In February 2019, the Microbial Genetic Resources Bank at INIA received fruits of V. corymbosum cv. 'Brigitta Blue' from Mariquina (-39.567869, -72.992461), located in the southern Chilean blueberry production zone, for post-harvest disease diagnosis. Asymptomatic fruits were incubated in moist-chambers at 25 °C with light/darkness cycles of 12 h. After 5 d, some fruits showed sunken areas and small surface wounds that exudated orange masses of conidia; under the epidermis, gray acervuli were also detected. After 15d, these fruits became dehydrated, mummified, and covered by mycelia, all characteristic symptoms of anthracnose (Wharton and Schilder 2008). In Chile, Colletotrichum gloeosporioides has, thus far, been the only causal agent of anthracnose reported in blueberry (Lara et al. 2003). Conidia exudated from the diseased fruit were inoculated on potato-dextrose agar (PDA) and incubated at 25 °C for 7 d. The resulting colony was predominantly cottony with gray aerial mycelium, displaying masses of pale orange conidia; on the reverse side, the colony was a pink-reddish color. Under a microscope, conidia were hyaline, fusiform to elliptic in shape, and displaying guttulate of 12.2±1.2 × 4.17±0.3 μm (n=30), characteristics coinciding with those described for Colletotrichum fioriniae (Pennycook 2017; Shivas and Tan 2009) (Supplementary Figure 1). The isolate was deposited in the Chilean Collection of Microbial Genetic Resources (CChRGM) as RGM 3330. Genomic DNA extraction of RGM 3330 and phylogenetic analyses were carried out according to Cisterna-Oyarce et al. (2022). A multi-locus sequencing analysis was carried out using five genetic markers. The internal transcribed spacer (ITS), glyceraldehyde 3-phosphate dehydrogenase (gapdh), actin (act), and chitin synthase 1 (chs-1) were PCR-amplified following Damm et al. (2012) and beta-tubulin (tub) following Glass and Donaldson (1995). Sequences were deposited in GenBank (ON364141 for ITS and ON369167-70 for tub, act, chs-1, and gapdh, respectively) (Sayers et al. 2021). A BLAST analysis carried out in SequenceServer (Priyam et al. 2019), using a custom database of sequences retrieved from Damm et al. (2012) and Liu et al. (2020), showed that all genetic markers were 100% identical to those of C. fioriniae CBS 128517T (ITS (540/540 identities), gapdh (249/249), act (245/245), and chs-1 (274/274)), except for tub, which shared 99.8% of its identities (416/417) with this species. Maximum likelihood phylogenetic estimation clustered RGM 3330 with C. fioriniae strains CBS 128517T and CBS 126526 with 100% bootstrap support (Supplementary Figure 1). Koch’s postulates were carried out with asymptomatic fruits of V. corymbosum cv. 'Brigitta Blue'. Prior to inoculation, fruits were surface-sterilized for 10 s in 70% ethanol, 3 s in 1% NaOCl, 10 s in 70% ethanol, rinsed three times with sterile distilled water, and subsequently placed in moist-chambers. Two groups of three repetitions of 20 fruits each were sprayed with 9 × 106 conidia/mL of RGM 3330 for the first group and with sterile distilled water for the control. After 5 d at 25 °C with light/darkness cycles of 12 h, only fruits sprayed with the conidial solution developed symptoms of anthracnose and the re-isolated fungi were identical in morphology to RGM 3330. This is the first report of C. fioriniae in blueberry in Chile. References: Cisterna-Oyarce, V., Carrasco-Fernández, J., Castro, J. F., Santelices, C., Muñoz-Reyes, V., Millas, P., Buddie, A. G., and France, A. 2022. Gnomoniopsis smithogilvyi: identification, characterization and incidence of the main pathogen causing brown rot in postharvest sweet chestnut fruits (Castanea sativa) in Chile. Australasian Plant Disease Notes 17:2. Damm, U., Cannon, P. F., Woudenberg, J. H., and Crous, P. W. 2012. The Colletotrichum acutatum species complex. Stud. Mycol. 73:37-113. Glass, N. L., and Donaldson, G. C. 1995. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl. Environ. Microbiol. 61:1323-1330. Lara, O., Velazquez, C. G., and Ascencio, C. 2003. Colletotrichum gloeosporiodes in blueberry fruit. in: XIII Congreso de Fitopatología. Liu, X., Zheng, X., Khaskheli, M. I., Sun, X., Chang, X., and Gong, G. 2020. Identification of Colletotrichum species associated with blueberry anthracnose in Sichuan, China. Pathogens 9:718. Pennycook, S. 2017. Colletotrichum fioriniae comb. & stat. nov., resolving a nomenclatural muddle. Mycotaxon 132:149-152. Priyam, A., Woodcroft, B. J., Rai, V., Moghul, I., Munagala, A., Ter, F., Chowdhary, H., Pieniak, I., Maynard, L. J., Gibbins, M. A., Moon, H., Davis-Richardson, A., Uludag, M., Watson-Haigh, N. S., Challis, R., Nakamura, H., Favreau, E., Gómez, E. A., Pluskal, T., Leonard, G., Rumpf, W., and Wurm, Y. 2019. Sequenceserver: a modern graphical user interface for custom BLAST databases. Mol. Biol. Evol. 36:2922-2924. Sayers, E. W., Cavanaugh, M., Clark, K., Pruitt, K. D., Schoch, C. L., Sherry, S. T., and Karsch-Mizrachi, I. 2021. GenBank. Nucleic Acids Res. 49:D92-D96. Shivas, R. G., and Tan, Y. P. 2009. A taxonomic re-assessment of Colletotrichum acutatum, introducing C. fioriniae comb. et stat. nov. and C. simmondsii sp. nov. Fungal Divers. 39:111-122. Wharton, P., and Schilder, A. 2008. Novel infection strategies of Colletotrichum acutatum on ripe blueberry fruit. Plant Pathol. 57:122-134.
Prospection of the endosphere of the native plant Fragaria chiloensis subsp. chiloensis f. patagonica from the foothills of the Chilean Andes led to the isolation of two strains of the genus Coniochaeta. We addressed the taxonomic placement of these strains based on DNA sequencing data using the ITS and LSU genetic markers, morphological features, and biochemical traits. One of these strains was identified as Coniochaeta hansenii, for which the anamorph and teleomorph states were described. The second strain did not seem to match any of the currently described species of this genus; therefore, we propose the name Coniochaeta fragariicola sp. nov.
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