Isolated Microspore Cultures of Triticale (× Triticosecale Wittm.)

Microspore embryogenesis (ME) is the shortest route to total homozygosity, as it results in the production of doubled haploid (DH) plants. However, the huge theoretical potential of this method is only rarely realized in practice, as there are no effective protocols available for a large number of crop species. In addition, the effectiveness of DH plant production is genotype dependent and highly variable not only between different species, but also between different cultivars, lines, and even individuals. The majority of modern triticale breeding materials are moderately responsive to ME induction. Therefore, procedures are still being developed and optimized for the requirements of specific genotypes. Based on the hypothesis of the important role of redox homeostasis in effective ME induction, we describe here a protocol modified by the application of reduced glutathione (GSH) as a factor that maintains microspore viability and promotes proper embryo-like structure (ELS) development in triticale.