A spectrophotometric assay for robust viability testing of seed batches using 2,3,5-triphenyl tetrazolium chloride: Using Hordeum vulgare L. as a model

Laura Lopez Del Egido*, David Navarro-Miró, Victor Martinez-Heredia, Peter E. Toorop, Pietro P.M. Iannetta

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)

Abstract

A comparative analysis was carried out of published methods to assess seed viability using 2,3,5-triphenyltetrazolium chloride (TTC) based assays of seed batches. The tests were carried out on seeds of barley (Hordeum vulgare cv. Optic) as a model. We established that 10%[w/v] trichloroacetic acid (TCA)/methanol is superior to the acetone and methanol-only based methods: allowing the highest recovery of formazan and the lowest background optical density (OD) readings, across seed lots comprising different ratios of viable and dead seeds. The method allowed a linear-model to accurately capture the statistically significant relationship between the quantity of formazan that could be extracted using the method we developed and the seed temperature-response, and seed viability as a function of artificially aged seed lots. Other quality control steps are defined to help ensure the assay is robust and these are reported in a Standard Operating Procedure.

Original languageEnglish
Article number747
JournalFrontiers in Plant Science
Volume8
DOIs
Publication statusPublished - 16 May 2017
Externally publishedYes

Keywords

  • Formazan
  • Hordeumvulgare
  • Seed viability
  • Seed-testing
  • Tetrazolium
  • TTC

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