sRNA-seq pf S.polyrhiza clone 9509 transiently expressing hpScarlet 12dpi
For Agrobacterium-mediated transient expression in Spirodela, Agrobacterium strain EHA105 was used. Prior to agroinfiltration, fronds were pretreated as in DOI: 10.1007/s11103-018-0778-x. . Following pretreatment, fronds were infiltrated with the Vir-induced Agrobacterium solution. For small RNA extraction from total RNA and TraPR input fraction, 10g of total RNA (as described above) or RNA extracted as described above from TraPR inputs was separated on a 15% denaturing polyacrylamide-urea gel. For size reference, 20l of microRNA marker (New England Biolabs, #N2102S) were loaded in the same gel. Following gel staining with peqGreen (Peqlab, #732-3196), and using the marker as reference, small RNAs ranging from 17-to-25nt were excised and recovered using the ZR small-RNA PAGE Recovery Kit (Zymo Research, #R1070).TraPR: Purifucation of Argonaute-loaded small RNAs was performed according to the protocol described in  DOI: 10.1093/nar/gkaa472. Libraries were prepared according to Hayashi et al., 2016 (DOI: 10.1038/nature20162).