MycoRed: Betalain pigments enable in vivo real-time visualisation of arbuscular mycorrhizal colonisation

PLoS Biol. 2021 Jul 14;19(7):e3001326. doi: 10.1371/journal.pbio.3001326. eCollection 2021 Jul.

Abstract

Arbuscular mycorrhiza (AM) are mutualistic interactions formed between soil fungi and plant roots. AM symbiosis is a fundamental and widespread trait in plants with the potential to sustainably enhance future crop yields. However, improving AM fungal association in crop species requires a fundamental understanding of host colonisation dynamics across varying agronomic and ecological contexts. To this end, we demonstrate the use of betalain pigments as in vivo visual markers for the occurrence and distribution of AM fungal colonisation by Rhizophagus irregularis in Medicago truncatula and Nicotiana benthamiana roots. Using established and novel AM-responsive promoters, we assembled multigene reporter constructs that enable the AM-controlled expression of the core betalain synthesis genes. We show that betalain colouration is specifically induced in root tissues and cells where fungal colonisation has occurred. In a rhizotron setup, we also demonstrate that betalain staining allows for the noninvasive tracing of fungal colonisation along the root system over time. We present MycoRed, a useful innovative method that will expand and complement currently used fungal visualisation techniques to advance knowledge in the field of AM symbiosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Betalains / metabolism*
  • Genes, Fungal
  • Genetic Markers
  • Medicago truncatula / microbiology
  • Mycorrhizae / genetics
  • Mycorrhizae / growth & development*
  • Mycorrhizae / metabolism
  • Nicotiana / genetics
  • Nicotiana / microbiology
  • Plant Roots / microbiology
  • Promoter Regions, Genetic
  • Symbiosis / genetics

Substances

  • Genetic Markers
  • Betalains

Grants and funding

This work was supported by an iCASE BBSRC-DTP (RG88096) sponsored by Coca-Cola to S.F.B & A.T., Gatsby Charitable Foundation (GAT3395/GLD) and Royal Society (UF110073, UF160413) to S.S., and a Natural Environmental Research Council Independent Research Fellowship (NE/K009303) to S.F.B. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.