Morex RNA-Seq 2010
Diverse Transcriptome of Morex Barley
This is one of several information resources developed in a project entitled, "Advancing the Barley Genome", supported by the Agriculture and Food Research Initiative of USDA's National Institute of Food and Agriculture. The Timothy J Close lab at UC Riverside produced total RNA from each of three sources of high-complexity RNA Morex barley tissues: roots from unstressed etiolated seedlings, shoot apices less than 10 mm in length, and differentiating spikes from 10 mm to white anther stage. These provided two blended-RNA samples (root, apices/spikes). The Gary J Muehlbauer lab at U Minnesota produced six Morex RNA samples from emerged spikes after inoculation with Fusarium graminearum (1 and 3 days) and seedling leaves inoculated with leaf rust (Puccinia hordei), Septoria speckled leaf blotch (Septoria passerinii Sacc.), spot blotch (Cochliobolus sativus) or bacterial blight (Xanthomonas transluscens). These provided two RNA samples (spikes, leaves). The Roger P Wise lab at Iowa State University provided one RNA sample by blending equal quantities of six RNA samples. These were from leaves inoculated with Blumeria graminis f. sp. hordei sampled at four different times after inoculation (2, 8, 16 and 32 hr) or leaves inoculated with barley stripe mosaic virus and then sampled at two different times after inoculation (6, 11 days). Libraries were prepared from each of the five mixed-RNA samples, individually bar-coded with unique adaptors. The five libraries were mixed in equal portions and applied to one lane of a flow cell for Illumina GAII sequencing. Library construction and sequencing were performed by Ambry Genetics (Aliso Viejo, CA) in June and July 2010, respectively. Sequences were demultiplexed and then assembled using Velvet/Oases by Steve Wanamaker at UC Riverside. The assembled sequences were made available for BLAST at harvest-blast.org and download from harvest-web.org/utilmenu.wc.