Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor

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 % Evans blue and the other staining with 1% 3-[4,5-dimethyltiazol-2-yl]-2,5-d...

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Autores principales:	Fernandez-Da Silva,Rafael, Menéndez-Yuffá,Andrea
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2006
Materias:	cell suspensions cell viability coffee Evans blue MTT protoplasts
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500016
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spelling	oai:scielo:S0717-345820060005000162007-01-04Viability in protoplasts and cell suspensions of Coffea arabica cv. CatimorFernandez-Da Silva,RafaelMenéndez-Yuffá,Andrea cell suspensions cell viability coffee Evans blue MTT protoplasts 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 % 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 37ºC. 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.9 n.5 20062006-10-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500016en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cell suspensions cell viability coffee Evans blue MTT protoplasts
spellingShingle	cell suspensions cell viability coffee Evans blue MTT protoplasts Fernandez-Da Silva,Rafael Menéndez-Yuffá,Andrea Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
description	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 % 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 37ºC. 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.
author	Fernandez-Da Silva,Rafael Menéndez-Yuffá,Andrea
author_facet	Fernandez-Da Silva,Rafael Menéndez-Yuffá,Andrea
author_sort	Fernandez-Da Silva,Rafael
title	Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
title_short	Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
title_full	Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
title_fullStr	Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
title_full_unstemmed	Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor
title_sort	viability in protoplasts and cell suspensions of coffea arabica cv. catimor
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2006
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500016
work_keys_str_mv	AT fernandezdasilvarafael viabilityinprotoplastsandcellsuspensionsofcoffeaarabicacvcatimor AT menendezyuffaandrea viabilityinprotoplastsandcellsuspensionsofcoffeaarabicacvcatimor
_version_	1718441759975931904

Viability in protoplasts and cell suspensions of Coffea arabica cv. Catimor

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