E-MTAB-682:E2
Transcription profiling by high throughput of Blumeria gramis during infection of Barley
Transcription profiling by high throughput of Blumeria gramis during infection of Barley
E2
Blumeria graminis f. sp. hordei (strain DH14) was maintained on barley (Hordeum vulgare) cv Golden Promise in an isolated, growth chamber maintained at 25C and a 16 hour light/ 8 hour dark cycle. The barley seedlings (10 to 12 day old) were inoculated by dusting conidia from heavily infected plants. Conidia were collected by vacuum aspiration onto Millipore filters (HATF02500) and frozen at -80C. Epiphytic structures were dissected by dipping infected leaves in a cellulose acetate (5%) dissolved in acetone; the acetone was left to dry off and the epiphytic material embedded in the cellulose acetate was dissected and snap frozen in liquid nitrogen and stored at -80C. Infected abaxial epidermis was collected by dissection, after removal of the epiphytic material, and stored at -80C. Primary barley leaves infected with B. graminis 5 days after infection were dissected to separate the epiphytic structures (samples E-1 to -3: surface runner hyphae, appressoria, condiophore foot cells, conidiophores and conidia, Fig. 1G) from the infected epidermis (samples H-1 to -3 containing haustoria). Three fully independent biological replicates where sampled. RNA was extracted from conidia and dissected material collected and ground in liquid nitrogen as described above. RNA was extracted as previously described by M. Both et al., Mol Plant-Microbe Interact 18, 125 (2005) and further purified using RNAClean XP beads (Beckman Coulter, Brea, CA, USA).
Imperial College IoSSB Sequencing Facility
E-MTAB-682:E2
Experimental Factor: ORGANISMPART
epiphytic