Hostname: page-component-586b7cd67f-l7hp2 Total loading time: 0 Render date: 2024-11-24T11:07:00.053Z Has data issue: false hasContentIssue false
  • English
  • Français

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
Get access

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Sihachakr D., Protoplastes: isolement, culture, régénération et fusion au polyéthylène glycol, in: Haïcour R., Biotechnologies végétales, Lavoisier Tec et Doc, Techn. Lab., Paris, France, 2002, pp. 5–328.
Matsumoto, K., Vilarinhos, A.D., Oka, S., Somatic hybridization by electrofusion of banana protoplasts, Euphytica 125 (2002) 317–324. CrossRef
Assani, A., Chabane, D., Haïcour, R., Bakry, F., Wenzel, G., Foroughi-Wehr, B., Protoplast fusion in banana (Musa spp.): comparison of chemical (PEG: polyethylene glycol) and electrical procedure, Plant Cell Tissue Organ Cult. 83 (2005) 145–151. CrossRef
Assani A., Haïcour R., Bakry F., Fouroughi-Wehr B., Les progrès réalisés dans la regénération des protoplastes de bananiers (Musa ssp.), in: Hamon S., Des modèles biologiques à l'amélioration des plantes, IRD Éd., Montpellier, France, 2001, pp. 193–206.
Galbraith, D.W., Microfluorometric quantitation of cellulose biosynthesis by plant protoplasts using calcofluor white, Physiol. Plant. 53 (1981) 111–116. CrossRef
Widholm, J.M., The use of fluorescein diacetate and phenosaphranin for determining the viability of cultured cells, Stain Technol. 47 (1972) 189–194. CrossRef
Megia, R., Haïcour, R., Tizroutine, S., Bui Trang, V., Rossignol, L., Sihachakr, D., Schwendiman, J., Plant regeneration from cultured protoplasts of the cooking banana cv. Bluggoe (Musa spp., ABB group), Plant Cell Rep. 13 (1993) 41–44. CrossRef
Bakry, F., Choix du matériel à utiliser pour l’isolement de protoplastes de bananier (Musa sp.), Fruits 39 (7–8) (1984) 449–452.
Assani, A., Haïcour, R., Wenzel, G., Fouroughi-Wehr, B., Bakry, F., Cote, F., Ducreux, G., Ambroise, A., Grapin, A., Influence of donor material and genotype on protoplast regeneration in banana and plantain cultivars (Musa spp.), Plant Sci. 162 (3) (2002) 355–362. CrossRef
Kao, K.N., Michayluk, M.R., Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media, Planta 126 (1975) 105–110. CrossRef
Morel, G., Wetmore, R.H., Fern tissue culture, Am. J. Bot. 38 (1951) 141–143. CrossRef
Haïcour R., Assani A., Matsumoto K., Guedira A., Banana protoplasts, in: Mohan Jain S., Swennen R., Banana improvement: Cellular, molecular biology, and induced mutations, Sci. Publ. Inc., Enfield, NH, USA, 2004, pp. 111–125
Haïcour R., Rossignol L., Bui Trang V., Megia R., Tizroutine S., Maîtrise de la culture et de la regénération des protoplastes de bananier, en vue de la création de nouvelles structures génétiques, in: Dubois J., Demarly Y., Quel avenir pour l'amélioration des plantes ?, J. Libbey Eurotext (Éd.), Actual. Sci., Paris, France, 1994, pp. 211–225.