RNA seq of triticum aestivum-FLW30

The transcriptome of FLW30 near isogenic line was investigated upon inoculation with the 46S119 pathotype. A total of 5 pots, including 5 mock-inoculated and 5 inoculated plants, were used with three biological replicates for each experiment. Uredinospores were collected from infected wheat plants and resuspended in sterile distilled water mixed with soltrol (20mg/100ml) for even distribution during seedling inoculation. The seedlings, at the two-leaf stage or flag leaves (approximately 20 days after sowing), were inoculated using an automizer and then kept in darkness at 10 degree C for 16 hours of light and 8 hours of darkness to maintain high humidity. Leaf samples were collected at three different time points: 12, 48, and 72hours post-inoculation (HPI). The collected samples were immediately preserved in RNA later and stored at -20 degree C until further use. RNA extraction from the leaf samples was performed using the Qiagen RNeasy Mini kit (Qiagen Inc, USA). The samples were then sequenced on a single HiSeq 4000 machine, generating at least 895 million 150bp paired-end reads per sample. Each sample represented a distinct experimental condition, including pathogen/mock, time points, and genotype.