The tolerance of the dune grass Leymus mollis (Triticeae; Poaceae) to various biotic and abiotic stresses makes it a very useful genetic resource for wheat breeding. Wide hybridization between L. mollis and wheat allows the introduction of Leymus chromosomes into the wheat genetic background and facilitates the integration of useful traits into wheat. However, the genetic basis controlling the physiological tolerance of L. mollis to multiple environmental stresses remains largely unexplored. Using suppression subtractive hybridization, we identified 112 osmotic-stress-responsive genes from L. mollis and confirmed their differential expression under osmotic stress. These genes were categorized into 13 functional categories, including cell defense and stress response, transcriptional regulation, signal transduction, biosynthesis of compatible solutes and cell wall metabolism. Representative genes were validated by northern blot and RT-PCR analyses of expression patterns in response to osmotic stress and abscisic acid treatment. The genes identified here represent a useful source of expressed sequence tags (ESTs) for the analysis and identification of Leymus chromosomes introduced into wheat. Furthermore, being highly conserved, genetically associated with osmotic stress tolerance and transferable to wheat, these ESTs provide significant tools for the development of EST-derived molecular markers for introgression of osmotic stress tolerance genes into wheat.
