Measurement of flux through sulfate assimilation using [35S]sulfate

Methods Enzymol. 2022;676:197-209. doi: 10.1016/bs.mie.2022.07.021. Epub 2022 Aug 13.


Sulfur metabolism provides a number of compounds that are essential for plant survival and fitness and that affect the yield and quality of crops. Sulfur metabolism is a dynamic process, responding to a number of external cues. Because of this dynamics and rapid turnover, steady-state levels of sulfur-containing compounds do not always fully reflect plant responses to such cues. Therefore, measurements of the flux through sulfate assimilation may give a more precise estimate of the effects of environmental stimuli or metabolic disturbances on sulfur metabolism. The flux can be determined after feeding plants with sulfate labelled with an isotopic tracer. Here we describe a protocol for using [35S]sulfate to measure flux through sulfate assimilation in Arabidopsis. The protocol can be adapted for any plant species and growth conditions, and does not require any special equipment beyond a standard high performance liquid chromatograph. We hope that the protocol will support our colleagues in a more frequent use of flux measurements to answer new biological questions in plant sulfur metabolism.

Keywords: Arabidopsis thaliana; Glutathione; Hydroculture; Radioactive tracer; Sulfate assimilation; Thiols; [(35)S]sulfate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis* / metabolism
  • Gene Expression Regulation, Plant
  • Plants / metabolism
  • Sulfates* / metabolism
  • Sulfur / metabolism


  • Sulfates
  • Sulfur