Measurement of Luciferase Rhythms in Soybean Hairy Roots

Methods Mol Biol. 2022:2398:65-73. doi: 10.1007/978-1-0716-1912-4_6.

Abstract

Firefly luciferase is widely used as a bioluminescence reporter, which is simple, high signal-to-noise ratio and especially suitable for the long-term analysis of circadian clock-regulated gene expression. Here, we report the method of tracking circadian rhythms in Agrobacterium rhizogenes-induced soybean hairy roots via TopCount™ Microplate Scintillation Counter or Deep-Cooled CCD camera. Using transgenic soybean hairy roots, we monitored the endogenous 24-h oscillations of clock genes expression and investigated the precise parameters of circadian rhythmicity. Researchers can easily analyze the circadian phenotype in legumes and non-legumes using bioluminescence reporters carried by the hairy roots, avoiding time-consuming transgenic work.

Keywords: Agrobacterium rhizogenes; Bioluminescence reporters; Circadian clock; Circadian rhythms; Firefly luciferase; Hairy roots; Soybean (Glycine max (L.) Merr.).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium
  • Glycine max* / genetics
  • Luciferases / genetics
  • Plant Roots / genetics
  • Plants, Genetically Modified / genetics

Substances

  • Luciferases

Supplementary concepts

  • Agrobacterium rhizogenes