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Chili pepper (
Capsicum
spp.) is a very important horticultural crop around the world and is especially important for Mexicans because of its impact in the culture and the cuisine. Biotechnological tools such as tissue culture techniques and specifically anther culture may be applied successfully for plant breeding and genetic improvement in order to generate isogenic lines (100% homozygous) in a shorter time in comparison with the classic breeding methods. In this chapter, a protocol for efficient recovery of chili pepper haploid plants from in vitro cultured anthers is described.