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ARS Home » Midwest Area » Peoria, Illinois » National Center for Agricultural Utilization Research » Mycotoxin Prevention and Applied Microbiology Research » Research » Publications at this Location » Publication #323682
Title: Rapid detection method for fusaric acid-producing species of Fusarium by PCR

Author
item LEE, THERESA - Rural Development Administration - Korea
item KIM, SOSOO - Rural Development Administration - Korea
item Busman, Mark
item Proctor, Robert
item HAM, HYEONHUI - Rural Development Administration - Korea
item LEE, SOOHYUNG - Rural Development Administration - Korea
item RYU, JAE-GEE - Rural Development Administration - Korea

Submitted to: Research in Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/23/2015
Publication Date: 12/31/2015
Citation: Lee, T., Kim, S., Busman, M., Proctor, R.H., Ham, H., Lee, S., Hong, S.K., Ryu, J.-G. 2015. Rapid detection method for fusaric acid-producing species of Fusarium by PCR. Research in Plant Disease. 21(4):326-329.

Interpretive Summary: Fungi typically cause two kinds of agricultural problems; they cause crop diseases and/or they produce toxic metabolites (mycotoxins) that pose health risks to humans and domestic animals. Most mycotoxins are synthesized by multiple enzymes that catalyze a series of biochemical reactions (a biosynthetic pathway) that transform relatively simple chemical compounds into more complex compounds. Genes that serve as the blue prints for the enzymes are typically located adjacent to one another along a chromosome in what is known as a biosynthetic gene cluster. Multiple species of the fungus Fusarium produce the mycotoxin fusaric acid, and a fusaric acid biosynthetic gene cluster has been identified in several species. In the current study, we used DNA sequence data from a gene in the cluster to develop a diagnostic assay to determine whether strains of Fusarium isolated from crops have the fusaric acid biosynthetic gene cluster and, therefore, the potential to produce the mycotoxin. We also combined this diagnostic assay with a previously developed assay for detection of a gene required for biosynthesis of another group of mycotoxins known as fumonisins. Thus, we have developed a diagnostic assay to determine the potential of strains of Fusarium to produce two types of mycotoxins. This research will be of use to plant pathologists, plant breeders, and other scientists involved in i) identification of fungi responsible for mycotoxin contamination problems and ii) development of new strategies to limit mycotoxin contamination in crop plants.

Technical Abstract: Fusaric acid is a mycotoxin produced by species of the fungus Fusarium and can act synergistically with other Fusarium toxins. In order to develop a specific detection method for fusaric acid-producing fungus, PCR prim¬ers were designed to amplify FUB10, a transcription factor gene in fusaric acid biosynthetic gene cluster. When PCR with Fub10-f and Fub10-r was performed, a single band (~550 bp) was amplified from F. oxysporum, F. proliferatum, F. verticillioides, F. anthophilum, F. bulbicola, F. circinatum, F. fujikuroi, F. redolens, F. sacchari, F. sub¬glutinans, and F. thapsinum, all of which were known for fusaric acid production. Whereas the FUB10 specific band was not amplified from Fusarium species known to be trichothecene producer. Because production of fusaric acid can co-occur in species that also produce fumonisin mycotoxins, we developed a multiplex PCR assay using the FUB10 primers as well as primers for the fumonisin biosynthetic gene FUM1. The assay yielded amplicons from fumonisin producers such as F. proliferatum and F. verticillioides, allowing for the simultaneous detection of species with the genetic potential to produce both types of mycotoxins.