A Protoplast-Based Bioassay to Quantify Strigolactone Activity in Arabidopsis Using StrigoQuant

Methods Mol Biol. 2021;2309:201-218. doi: 10.1007/978-1-0716-1429-7_16.

Abstract

Understanding the biological background of strigolactone (SL) structural diversity and the SL signaling pathway at molecular level requires quantitative and sensitive tools that precisely determine SL dynamics. Such biosensors may be also very helpful in screening for SL analogs and mimics with defined biological functions.Recently, the genetically encoded, ratiometric sensor StrigoQuant was developed and allowed the quantification of the activity of a wide concentration range of SLs. StrigoQuant can be used for studies on the biosynthesis, function and signal transduction of this hormone class.Here, we provide a comprehensive protocol for establishing the use of StrigoQuant in Arabidopsis protoplasts. We first describe the generation and transformation of the protoplasts with StrigoQuant and detail the application of the synthetic SL analogue GR24. We then show the recording of the luminescence signal and how the obtained data are processed and used to assess/determine SL perception.

Keywords: Arabidopsis; Biosensor; D14; Firefly; Luminescence; Plant hormones; Protoplasts; Ratiometric sensor; SMXL6; StrigoQuant; Strigolactone; Synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Biological Assay*
  • Biosensing Techniques*
  • Heterocyclic Compounds, 3-Ring / metabolism*
  • Lactones / metabolism*
  • Luciferases, Firefly / genetics
  • Luciferases, Firefly / metabolism
  • Luciferases, Renilla / genetics
  • Luciferases, Renilla / metabolism
  • Luminescent Measurements
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Protoplasts / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction

Substances

  • Arabidopsis Proteins
  • GR24 strigolactone
  • Heterocyclic Compounds, 3-Ring
  • Lactones
  • Recombinant Fusion Proteins
  • Luciferases, Renilla
  • Luciferases, Firefly