Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
Electron. j. biotechnol
;
9(5)Oct. 2006. ilus, graf
Article
Dans Anglais
| LILACS
| ID: lil-451683
ABSTRACT
Two 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 percent Evans blue and the other staining with 1 percent 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 37°C. The time of incubation was shown to be important, since longer times improved the reaction; the highest viability value was obtained after incubation for 120 min.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
langue:
Anglais
Texte intégral:
Electron. j. biotechnol
Thème du journal:
Biotechnologie
Année:
2006
Type:
Article
/
descriptif de projet
Pays d'affiliation:
Venezuela
Institution/Pays d'affiliation:
Universidad Central de Venezuela/VE
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