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Protoplast isolation and culture for banana regeneration via somatic embryogenesis

Published online by Cambridge University Press:  02 September 2009

Robert Haïcour ,
Akym Assani ,
Viet BuiTrang  and
Abdelkarim Guedira
Robert Haïcour
Affiliation:
Lab. Ecol. Syst. Evol., UMR 8079, Bât 362, Univ. Paris Sud XI, F- 91405 Orsay Cedex, France
Akym Assani
Affiliation:
Univ. Guelph, Dep. Plant Agric., Bovey Building, N1G 2W1, Guelph, Ontario, Canada
Viet BuiTrang
Affiliation:
Lab. Physiol. Vég., 227 Nguyen Van Cu, Q.5, Hô Chi Minh Ville, Viet Nam
Abdelkarim Guedira
Affiliation:
Dép. Biol., Lab. Physiol. Vég., Fac. Sci. Rabat, Univ. Mohammed V Agdal, Ave. Ibn Battouta, BP 1014, 1000 Rabat, Maroc
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Abstract

Introduction. This protocol describes a method for obtaining protoplasts from banana leaves, calli and cell suspensions, and their sustainable development via somatic embryogenesis from embryogenic cell suspensions. The principle, key advantages, starting plant material, time required and expected results are presented. Materials and methods. This part describes the required laboratory materials, and media preparation for protoplast production and culture. Results. The first protoplasts may be seen after 30 min of incubation in enzyme maceration. With protoplasts from embryogenic cell suspension, complete development into a whole plant, through somatic embryogenesis, is observed in 12 weeks. The first cell divisions occur on feeder layers 3–8 days after protoplast plating. Proembryo formation is observed 14–21 days after initiation of protoplast culture. The transfer of derived embryo plantlets, at 8–10 weeks after protoplast plating, onto growth regulator-free medium, leads to plant rooting and elongation.

Keywords

FranceMusa sp.methodculture mediaculture techniquesprotoplastssomatic embryogenesisin vitro regenerationFranceMusa sp.méthodemilieu de culturetechnique de cultureprotoplasteembryogenèse somatiquerégénération in vitro
Type
Research Article
Information
Fruits , Volume 64 , Issue 4 , July 2009 , pp. 261 - 269
Copyright
© CIRAD, EDP Sciences, 2009

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