Articles
ISOLATION AND CULTURE OF PROTOPLASTS FROM CELL SUSPENSIONS OF CACTUS PEAR (OPUNTIA FICUS-INDICA MILL.)
Article number
728_11
Pages
93 – 96
Language
English
Abstract
Tissue culture techniques are essential to facilitate the application of modern biotechnological methods to cactus pear genetic improvement.
Viable protoplasts from cell suspensions of Opuntia ficus-indica, were obtained by incubating cells in medium containing 1% cellulase, 0.5% pectinase, 0.1 mM CaCl2 and 0.7 M glycine, for 10 h in the dark at 28°C. After transferring protoplasts to a medium consisting of full-strength MS salts and vitamins, 0.7 M glucose, 0.5 mM MES, 0.9 µM kinetin, 1.0 µM picloram, 2.3 µM 2,4-D and 0.2 g/dm-3 casein hydrolysate, high levels of cell division as well as cell wall regeneration were observed.
Under the optimized conditions described here, we obtained a yield of 5 x 105 viable protoplasts for each cm3 of cell suspension and these isolated protoplasts were viable as estimated by Evans blue staining.
Viable protoplasts from cell suspensions of Opuntia ficus-indica, were obtained by incubating cells in medium containing 1% cellulase, 0.5% pectinase, 0.1 mM CaCl2 and 0.7 M glycine, for 10 h in the dark at 28°C. After transferring protoplasts to a medium consisting of full-strength MS salts and vitamins, 0.7 M glucose, 0.5 mM MES, 0.9 µM kinetin, 1.0 µM picloram, 2.3 µM 2,4-D and 0.2 g/dm-3 casein hydrolysate, high levels of cell division as well as cell wall regeneration were observed.
Under the optimized conditions described here, we obtained a yield of 5 x 105 viable protoplasts for each cm3 of cell suspension and these isolated protoplasts were viable as estimated by Evans blue staining.
Authors
R.M. Liamoca Zarate, J. Landsann, F.A.P. Campos
Keywords
forage crops, Opuntia ficus-indica, protoplast isolation
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