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.
CITATION STYLE
Lopez Del Egido, L., Navarro-Miró, D., Martinez-Heredia, V., Toorop, P. E., & Iannetta, P. P. M. (2017). A spectrophotometric assay for robust viability testing of seed batches using 2,3,5-triphenyl tetrazolium chloride: Using Hordeum vulgare L. as a model. Frontiers in Plant Science, 8. https://doi.org/10.3389/fpls.2017.00747
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