Barley anther culture

Abstract

The production of doubled haploid (DH) barley (Hordeum vulgare L.) plants has proven to be a highly valuable tool for plant breeding, allowing the release of a high number of new cultivars (Devaux et al., 1996). Doubled haploids have also been fundamental for genetic analysis, linkage maps production and QTL analysis in barley. Several anther culture protocols have been established for an efficient production of DH lines by different public institutions and private companies. However, the low embryogenic capacity and the high albinism rate of some genotypes are still open fields for further improvements in these protocols. To be successfully used in a breeding program, any particular protocol should produce a large number of DH lines from all the genotypes. During the last years, our aim has been focused on the establishment of an efficient protocol for the production of doubled haploid plants from a wide range of genotypes, as well as from F₁ crosses of putative great agronomic performance. The results achieved over five years (1993–1997) in our laboratory resulted in a gradual increase in the numbers of green plants per 100 anthers (from 6.3 up to 17.4), as well as DH fertile lines per 100 anthers (from 1.2 to 10.3). Major changes of the initial protocol have been the increase of mannitol concentration in the pretreatment medium from 0.3 to 1.0 M, the introduction of Ficoll in the induction medium, the optimization of the regeneration medium composition, and the procedure for transferring the plants to soil (Cistué et al., 1994, 1999; Castillo et al., 2000).