Several aspects of in vitro culture have potential for cereal improvement. This paper focuses on evaluation of somaclonal variation (SV) from immature embryo callus culture, and doubled haploid (DH) production via anther culture in barley. Genetically stable SV was observed for several seedling morphological traits such as albino, yellow, light green and lethal. SV occurred at approximately half the frequency of azide-induced mutagenesis. The potential for widespread application of anther culture-mediated DH production in barley breeding and genetic studies was increased through culture procedure improvements and understanding the inheritance of anther culture response. Methodology improvements included substitution of inexpensive gelrite for expensive ficoll or agarose, ability to grow anther donor plants under field as well as growth chamber conditions and flexibility in cold pretreatment/storage of anther donor spikes for 4-6 weeks prior to anther plating. From diallel analysis, inheritance of anther culture response was complex with additive and dominance effects for embryoid formation, total plant regeneration and green plant regeneration and reciprocal effects (maternal) for green plant regeneration. High x low responder crosses generated F1's that were intermediate in response and low x low crosses sometimes produced F1 heterosis for green plant regeneration. Therefore, some recalcitrant types appear to be usable in anther culture DH production systems within a breeding program.