A new and effective method to induce infection of Phyllachora maydis into corn for tar spot studies in controlled environments

José E. Solórzano, Shea E. Issendorf, Milton T. Drott, Jill C. Check, Emily M. Roggenkamp, C. D. Cruz, Nathan M. Kleczewski, Carlos C. Gongóra-Canul, Dean K. Malvick

Research output: Contribution to journalArticlepeer-review


Background: Tar spot of corn is a significant and spreading disease in the continental U.S. and Canada caused by the obligate biotrophic fungus Phyllachora maydis. As of 2023, tar spot had been reported in 18 U.S. states and one Canadian Province. The symptoms of tar spot include chlorotic flecking followed by the formation of black stromata where conidia and ascospores are produced. Advancements in research and management for tar spot have been limited by a need for a reliable method to inoculate plants to enable the study of the disease. The goal of this study was to develop a reliable method to induce tar spot in controlled conditions. Results: We induced infection of corn by P. maydis in 100% of inoculated plants with a new inoculation method. This method includes the use of vacuum-collection tools to extract ascospores from field-infected corn leaves, application of spores to leaves, and induction of the disease in the dark at high humidity and moderate temperatures. Infection and disease development were consistently achieved in four independent experiments on different corn hybrids and under different environmental conditions in a greenhouse and growth chamber. Disease induction was impacted by the source and storage conditions of spores, as tar spot was not induced with ascospores from leaves stored dry at 25 ºC for 5 months but was induced using ascospores from infected leaves stored at -20 ºC for 5 months. The time from inoculation to stromata formation was 10 to 12 days and ascospores were present 19 days after inoculation throughout our experiments. In addition to providing techniques that enable in-vitro experimentation, our research also provides fundamental insights into the conditions that favor tar spot epidemics. Conclusions: We developed a method to reliably inoculate corn with P. maydis. The method was validated by multiple independent experiments in which infection was induced in 100% of the plants, demonstrating its consistency in controlled conditions. This new method facilitates research on tar spot and provides opportunities to study the biology of P. maydis, the epidemiology of tar spot, and for identifying host resistance.

Original languageEnglish (US)
Article number83
JournalPlant Methods
Issue number1
StatePublished - Dec 2023

Bibliographical note

Funding Information:
We thank the Minnesota Invasive Terrestrial Plants and Pests Center for supporting this research. We thank, Dr. Pablo Olivera, Dr. Cory Hirsch, Crystal Floyd, Alexis Shatrau, and Olivia Anderson from the Department of Plant Pathology at the University of Minnesota, Dr. Candice Hirsch and Michael Burns from the Agronomy and Plant Genetics Department at the University of Minnesota, and Douglas Brinkman and Tha Cha from the University of Minnesota Plant Growth Facilities for technical assistance with this research. We also thank Dr. Vinson Doyle from the Department of Plant Pathology at Louisiana State University for discussing ideas for this study.

Funding Information:
This study was funded by the Minnesota Invasive Terrestrial Plants and Pest Center (Primary project # 00050727).

Publisher Copyright:
© 2023, BioMed Central Ltd., part of Springer Nature.


  • Biotroph
  • Black spot
  • Disease
  • Fungus
  • Inoculation
  • Maize
  • Method
  • Replicable

PubMed: MeSH publication types

  • Journal Article


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