Dear Pooja and Manoj,
Nice protocol! It worked perfectly for Arabidopsis Col-0 (Columbia), it is fairly easy to distinguish between viable and non-viable seeds, and few seeds have intermediate staining.
However, other accessions such as Ws (Wassilewskija), show high heterogeneity with many partially stained seeds, even for controls, and it becomes very difficult to distinguish between viable and non-viable seeds. This has already been described in some publications since Ws seed coat seems to be slightly permeable for tetrazolium.
I have tried to improve this staining by washing with a scarification solution (20 ml commercial bleach and 100 µl Triton X-100 in 100 ml autoclaved distilled water), or increasing incubation time, but was unsuccessful. Any recommendations to improve this assay? Thank you very much for reading me.
4/28/2021 7:44:10 AM Reply