Identifying genes in Fusarium verticillioides through forward and reverse genetics

J. B. Ridenour, R. L. Hirsch, B. H. Bluhm

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

6 Scopus citations


The increasing availability of sequenced genomes for plant pathogenic fungi has revolutionized molecular plant pathology in recent years. However, the genetic regulatory networks underlying many important components of pathogenesis remain poorly defined. Although the protocols outlined in this chapter can be utilized to identify genes regulating a wide range of biological processes in many filamentous fungi, we focus on describing how to identify genes through forward and reverse genetics, using the plant pathogenic fungus Fusarium verticillioides as a model for the protocol. Specifically, this chapter explains how to create a collection of insertional mutants via Restriction Enzyme Mediated Integration (REMI) and how to screen mutants with a high-throughput method to visualize defects in amylolysis. Next, techniques are described to define the genomic lesions in REMI mutants with genome-walker PCR in order to identify candidate genes. Finally, protocols are presented describing a reverse-genetic approach to disrupt candidate genes in the wild-type strain with a split-marker strategy to confirm the phenotype observed in the REMI mutant.

Original languageEnglish
Title of host publicationPlant Fungal Pathogens
Subtitle of host publicationMethods and Protocols
Number of pages23
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Amylolysis
  • Forward genetics
  • Fusarium verticillioides
  • Protoplasts
  • Reverse genetics
  • Transformation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


Dive into the research topics of 'Identifying genes in Fusarium verticillioides through forward and reverse genetics'. Together they form a unique fingerprint.

Cite this