RNA-seq analysis in callus of barley

We have been identified three loci responsible for transformation amenability in barley, and we are investigating the genes for this phenotype which may be associated with callus induction, callus growth, Agrobacterium-barley interaction, green shoot regeneration and rooting. In this project, RNA-seq analysis was performed to isolate the genes involved in callus induction/growth and green shoot regeneration from callus in barley. The RNA was extracted from the calli of Golden Promise and Haruna Nijo which were transformation amenable and recalcitrant barley cultivars, respectively, as well as from the transgenic calli expressing baby boom (BBM) or Wuschel 2 (WUS2) genes regulated under the maize ubiquitin promoter.