Abstract
A protocol for the axillary bud proliferation of prickly pear (Opuntia; Cactaceae) is presented. This genus is widely used as a crop in the arid and semi-arid areas of the globe worldwide, providing numerous benefits for human and animal consumption. In vitro culture for axillary bud proliferation is of great use to obtain a large quantity of plants in a short period of time, with potential uses in production and for the preservation of endangered species of the Opuntia genus.The optimal medium for Opuntia in vitro culture consists of Murashige and Skoog medium (MS) and L2 vitamins. To increase the yield of the axillary bud proliferation, we recommend the addition of plant growth regulators (PGRs). This work suggests a 15 d incubation in the medium with 2.2 mg/L of benzyl aminopurine (BA) after which the explants are transferred to the medium without PGRs. We explain as well how to adapt the plant to ex vitro conditions.
Keywords: Prickly pear, In vitro culture, Axillary bud proliferation, Ex vitro adaptation
Background
The genus Opuntia (prickly pear) is one of the members of the Cactaceae family (Bravo-Hollis, 1978). Although it is native to the Americas, it currently grows in the wild and commercial plantations of the South of Europe, North of Africa, Australia, Middle East, West Asia and other regions of the world (Ochoa and Barbera, 1995; Kiesling and Metzing, 2017). Prickly pear has profound effects on the arid and semi-arid environments as well as the human communities that live in those areas because of its high biomass yields despite growing in drylands (Acevedo et al., 1983). In many of such areas, the genus Opuntia is exploited for a number of things. The young cladodes of Opuntia can be consumed as a vegetable; the fruit is eaten directly or processed into jelly, juice or sweets (Barba et al., 2017). The whole plant can be used for animal forage, living fence (Las Casas et al., 2017) and recently it started to be used commercially to produce biofuel (Aké Madera, 2014). In addition, the Opuntia genus has a number of bioactive and nutritionally valuable compounds (Betancourt et al., 2017; Melgar et al., 2017).The advantages of in vitro culture include high proliferation rates and the production of pathogen-free plants (Shedbalkar et al., 2010). In their native environments, some Opuntia species are suffering losses in their populations, due to exploitation without replacement that has been carried out traditionally (Rocha-Flores et al., 2017). In vitro culture thus brings the possibility of recovering the populations of endangered plants (Torres-Silva et al., 2018), as well as a large scale production for agriculture. Here we present a protocol for plant in vitro culture of the Opuntia genus that includes the cytokinin benzyl aminopurine (BA).
Materials and Reagents
Equipment
Procedure
Notes
Recipes
Acknowledgments
The protocol presented here was developed and optimized thanks to the funding of Universidad de Guadalajara by means of the P3E program. We thank Cátedras-CONACYT program for financial support. Authors declare no conflict of interest.
References
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