Freshly harvested barley seeds are considered as dormant since they do not germinate at temperatures above 20 degrees C. This dormancy is broken during dry storage. Molecular regulation of dormancy was investigated using cDNA-AFLP to identify transcripts differentially expressed in dormant and non-dormant embryos. Transcript patterns in embryos from dry dormant and non-dormant seeds and from both seeds imbibed for 5 h at 30 degrees C, a temperature at which dormancy is expressed, were compared. Thirty-nine Transcript-Derived Fragments (TDF) that were reproducibly differentially expressed among treatments were identified, and 25 of these were cloned and sequenced. Among these, eight transcripts were observed to be differentially expressed during after-ripening, seven of which decline, probably due to post-maturation degradation. HV13B, TDF identified as having homology to fructose-6-phosphate-2-kinase/fructose-2,6-biphosphatase, may have a role in the maintenance of dormancy in barley and probably in other cereals. During the first 5 h of imbibition, there was expression of 24 TDF which was apparently independent of dormancy, revealing putative epigenetic regulation. This was typified by HV44A, a SET domain protein. Seven TDF differentially expressed, and especially HV12D, HV42B, and HV32B, in dormant and non-dormant seeds were potential signalling elements. HV12D had homology with an ARIADNE gene which could be implicated in ABA signalling.