Engelmann Florent, Malaurie Bernard, N'Nan Oulo
Institut de recherche pour le développement (IRD), UMR DIAPC, 911 Avenue Agropolis, BP 64501, 34394, Montpellier Cedex 5, France.
Methods Mol Biol. 2011;710:63-72. doi: 10.1007/978-1-61737-988-8_6.
Coconut is a very important crop for millions of people in tropical countries. With coconut, in vitro culture protocols have been developed with two main objectives, viz. the large scale production of particular types of coconuts and the international exchange and conservation of coconut germplasm. The methods described in this chapter have been developed in the framework of collaborative activities between research institutes in Côte d'Ivoire and France. Two coconut embryo in vitro collecting protocols have been established, one consisting of storing the disinfected embryos in a KCl solution until they are brought back to the laboratory, where they are re-disinfected and inoculated in vitro under sterile conditions, and the other including in vitro inoculation of the embryos in the field. For international germplasm exchange, zygotic embryos inoculated in vitro in plastic test tubes or endosperm cylinders containing embryos in plastic bags are used. For in vitro culture, embryos are inoculated on semi-solid medium supplemented with sucrose and activated charcoal and placed in the dark, and then transferred to light conditions with the same (solid or liquid) medium once the first true leaf is visible and the root system has started developing.
椰子对热带国家的数百万人来说是一种非常重要的作物。对于椰子,已经制定了离体培养方案,主要有两个目标,即大规模生产特定类型的椰子以及椰子种质的国际交流与保存。本章所述方法是在科特迪瓦和法国的研究机构之间的合作活动框架内开发的。已经建立了两种椰子胚离体采集方案,一种是将消毒后的胚保存在氯化钾溶液中,直到带回实验室,在实验室中再次消毒并在无菌条件下进行离体接种,另一种包括在田间对胚进行离体接种。对于国际种质交换,使用在塑料试管中离体接种的合子胚或装在塑料袋中含有胚的胚乳圆柱体。对于离体培养,将胚接种在添加了蔗糖和活性炭的半固体培养基上,置于黑暗中,一旦第一片真叶可见且根系开始发育,就转移到相同(固体或液体)培养基的光照条件下。