Small RNA sequencing of PEBV-infected N. benthamiana plants

To test if BYDV 17K and its phosphor-mimic mutant 17K5D may differ in regulating vsiRNA accumulation in virus-infected plant cells, the coding sequence of 17K or 17K5D was introduced into the genome of pea early browning virus (PEBV) to produce the recombinant virus i.e., PEBV-17K and PEBV-17K5D. PEBV is a bipartite RNA virus with its full-length infectious clones capable of infecting tobacco. The N. benthamiana plants were inoculated with the PEBV-GFP (expressing free GFP), PEBV-17K or PEBV-17K5D by agroinfiltration for developing three type of virus-infected tobacco plants. The small RNAs were extracted from the above plants at 14 dpi and were used to construct the small RNA-Seq library.