Visualization of Arbuscular Mycorrhizal Fungal Extraradical Hyphae and Spores Vitality and Activity

Methods Mol Biol. 2020:2146:61-71. doi: 10.1007/978-1-0716-0603-2_6.

Abstract

The hyphae and spores of arbuscular mycorrhizal (AM) fungi represent an essential component in the extraradical zone due to their role in nutrients and water uptake and as propagules that allow the perpetuation of the AM symbiosis over time, respectively. However, the attention of scientific literature is usually more focused on root colonization than on the study of the extraradical components of AM fungi, especially their vital, active, or functional fractions. This chapter presents some easy-to-use alternatives for staining vital, active, or functional structures of AM fungi for their subsequent microscopic visualization, such as the application of enzyme-based stains, NADPH formation, and also nucleus staining. Some modified methods for the extraction of mycelium from the soil are also presented.

Keywords: Confocal laser scanning microscopy; Dehydrogenase activity; Gridline intersection; Hyphae density; Mycelia and spores isolation; Vital staining.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Hyphae / growth & development*
  • Hyphae / ultrastructure
  • Mycelium / genetics
  • Mycelium / growth & development
  • Mycorrhizae / growth & development*
  • Mycorrhizae / ultrastructure
  • Plant Roots / microbiology
  • Plant Roots / ultrastructure
  • Spores, Fungal / growth & development
  • Spores, Fungal / ultrastructure
  • Staining and Labeling / methods*
  • Symbiosis*
  • Water / chemistry

Substances

  • Water