Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
Financial support: Project S1-98003209 FONACIT (Ministerio de Ciencia y Tecnología de Venezuela).
Keywords: cell suspensions, cell viability,coffee, Evans blue, MTT, protoplasts.
methods were evaluated in order to assess the viability of protoplasts
and cell suspensions of Coffea arabica cv. Catimor used in
a protocol of transformation by electroporation. One method consisted
of staining with 1 % Evans blue and the other staining with 1% 3-[4,5-dimethyltiazol-2-yl]-2,5-diphenyltetrazolium
bromide (MTT). When Evans blue was applied to viable cells and protoplasts
they either did not stain or acquired a faint blue colour. However,
it was difficult to distinguish the non-viable cells in the wide spectrum
of clear blue tonalities. In contrast, with the MTT assay only the
viable cells and protoplasts reduced this salt to the red coloured
formazan; viable and non-viable cells were distinguished more clearly
with MTT than with Evans blue. The optimal temperature for the reaction
with MTT was
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