The authors thank Amy McGovern and Crystal Probyn for skilled technical assistance. Van Dam P, de Sain M, ter Horst A, van der Gragt M, Rep M (2018) Use of comparative genomics-based markers for discrimination of host specificity in Fusarium oxysporum. O’Donnell K, Ward TJ, Robert VARG, Crous PW, Geiser DM, Kang S (2015) DNA sequence-based identification of Fusarium: current status and future directions. Geiser DM, del Mar Jiménez-Gasco M, Kang S, Makalowska I, Veeraraghavan N, Ward TJ et al (2004) FUSARIUM-ID v 10: a DNA sequence database for identifying Fusarium. O’Donnell K, Rooney AP, Proctor RH, Brown DW, McCormick SP, Ward TJ et al (2013) RPB1 and RPB2 phylogeny support an early cretaceous origin and a strongly supported clade comprising all agriculturally and medically important fusaria. O’Donnell K, Sutton DA, Rinaldi MG, Sarver BAJ, Balajee SA, Schroers H-J et al (2010) Internet-accessible DNA sequence database for identifying fusaria from human and animal infections. O’Donnell K, Kistler HC, Cigelnik E, Ploetz RC (1998) Multiple evolutionary origins of the fungus causing Panama disease of banana: concordance evidence from nuclear and mitochondrial gene genealogies. Leslie JF, Summerell BA (2006) The Fusarium laboratory manual. Pennsylvania State University Press, University Park, PA Nelson PE, Toussoun TA, Marasas WFO (1983) Fusarium species: an illustrated manual for identification. Mitt Biol Bundesanst Land-Forstwirtsch 209:1–406 Gerlach W, Nirenberg HI (1982) The genus Fusarium – a pictorial atlas. Taylor JW, Jacobson DJ, Kroken S, Kasuga T, Geiser DM, Hibbett DS et al (2000) Phylogenetic species recognition and species concepts in fungi. Geiser DM, Al-Hatmi A, Aoki T, Arie T, Balmas V, Barnes I et al (2020) Phylogenomic analysis of a 55.1 kb 19-gene dataset resolves a monophyletic Fusarium that includes the Fusarium solani species complex. Wingfield MJ, De Beer W, Slippers B, Wingfield BD, Groenewald JZ, Lombard L et al (2011) One fungus, one name promotes progressive plant pathology. Here we describe how to obtain single-spored pure cultures from symptomatic host tissue and a molecular identification by querying publicly accessible DNA sequence databases using a portion of translation elongation factor 1-α ( TEF1), the largest subunit of RNA polymerase ( RPB1), and/or the second largest subunit of RNA polymerase ( RPB2). With over 300 phylogenetically distinct species, and a dearth of phenotypical characteristics, DNA sequence data in most instances is the only reliable means for obtaining an accurate species identification. In addition, head blight and ear rot diseases are associated with the accumulation of mycotoxins in cereals. Using this approach, the cost of analysis for abundance studies can be greatly reduced compared to DNA sequencing of each isolate.Fusarium ranks as the most important group of plant pathogens, responsible for a wide range of economically destructive diseases, including vascular wilts and root, crown, and stem rots. This method unambiguously identified 78% of the fungal taxa in this study and we were able to rapidly determine which isolates should be subjected to further analysis by DNA sequencing. In addition, isolates containing more than one species were identified, allowing isolates not in the database to be sequenced for further analysis. A database of Antarctic fungal electropherograms was produced containing each of the species and was used as the first step in a tiered system for species identification. We optimized template concentration and verified the effect of primer selection from eight commonly used fungal polymerase chain reaction primers on ARISA chromatographs for 46 fungal species commonly isolated from south Victoria Land. This method is well suited for the study of culturable Antarctic soil fungal communities where both abundance and diversity are relatively low. We utilized an automated ribosomal intergenic spacer analysis (ARISA) method as a more rapid alternative to classical morphological/nutritional identification and a less expensive alternative to sequencing for identification and grouping of isolates in culture-based fungal abundance studies.
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