Pure Culture and DNA Sequence-Based Identification of Fusarium from Symptomatic Plants and Diverse Substrates

Abstract

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. In addition, head blight and ear rot diseases are associated with the accumulation of mycotoxins in cereals. 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. 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).

Keywords: ABI sequence chromatogram; BLASTn query; DNA extraction; DNA sequencing; FUSARIUM-ID; Fusarium MLST; NCBI GenBank; PCR amplification; Pure culture.