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Effectiveness of three micropropagation techniques to dissociate cytochimeras in Musa spp
Authors:Roux  Nicolas  Dolezel  Jaroslav  Swennen  Rony  Zapata-Arias  Fransisco J.
Affiliation:(1) FAO/IAEA Agriculture and Biotechnology Laboratory, International Atomic Energy Agency Laboratories, Plant Breeding Unit, A-2444 Seibersdorf, Austria;(2) Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovska 6, CZ-77200 Olomouc, Czech Republic;(3) Laboratory of Tropical Crop Improvement, Katholieke Universiteit Leuven, Kardinaal Mercierlaan 92, B-3001 Heverlee, Belgium;(4) FAO/IAEA Agriculture and Biotechnology Laboratory, International Atomic Energy Agency Laboratories, Plant Breeding Unit, A-2444 Seibersdorf, Austria
Abstract:In vitro mutagenesis of multicellular meristems of Musa spp. leads to a high degree of chimerism. Repeated vegetative propagation must be carried out to dissociate chimeras but the minimum number of cycles required is unknown. In general, mutated cells are difficult to monitor but mutations which result in a change in genome number are an exception. We simulated this case by colchicine treatment, followed by flow cytometric analysis. Colchicine treatment induced ploidy chimerism (mixoploidy), and chimera dissociation was assessed using three different propagation systems (shoot-tip culture technique - ST, multi-apexing culture technique - MA and corm slice culture technique - CS). The average percentage of cytochimeras was reduced from 100% to 36% after three subcultures using shoot-tip culture, from 100% to 24% when propagating by the corm slice culture technique and from 100% to 8% after the same number of subcultures using the multi-apexing technique. All propagation systems failed to eliminate chimerism completely. Factors that may influence chimera dissociation in vitro are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.
Keywords:chimera  flow cytometry   in vitro tissue culture   Musa acuminata   mixoploidy  polyploidisation
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