Barley TET8 and two RNA helicases cooperatively regulate cross-kingdom RNAi and immunity to powdery mildew

The barley cultivar Golden Promise was inoculated with Bgh A6 isolate, and epidermal tissues were collected from five conditions: healthy leaves, and Bgh-infected leaves at 8, 16, 24, and 72 hpi. Total RNA was extracted using the TRIzol reagent as instruction. Small RNA fragments (18-28 nt) were separated and used to construct sequencing libraries, which were sequenced on the Illumina HiSeq 2000 platform (sRNA-seq). For degradome sequencing, two pooled libraries were prepared: one comprising barley leaf epidermis infected with Bgh A6 at 8, 16, 24, and 72 h, and the other consisting of 7-day-old Bgh conidia and infection structures (hyphae and appressoria) harvested at 3 and 5 dpi.