Fig. 10

Trafficking of Cy3 labeled wheat mir_17532_x1 mimics (miR17532) from wheat roots to C. rosea-GFP conidia and hyphae. A Representative confocal images showing the co-localization of Cy3-miR17532 (Magenta), C. rosea-GFP (Green), and merge (right panel). White arrows indicate germinated C. rosea conidia. B A representative confocal image (enlarged) shows the internalization of Cy3-miR17532 mimic (Magenta) by C. rosea conidia (Green) and submerge (right panel). After 24 h of incubation with miR17532, wheat roots were washed with 0.1M KCl and 0.01 M Triton X100 to remove surface-bound miRNA oligos. Conidia from C. rosea-GFP were applied to the roots, and Cy3 fluorescence was determined t72 hpi. C–E Detection and quantification of wheat miRNAs mimic inside wheat roots using stem-loop-RT-qPCR (CP. infestans miR8788 mimic miR8788; D wheat mir_17532_x1 mimic miR17532; E wheat mir_12061_x13 mimic miR12061). F RT-qPCR showing the expression of the pks29 gene in C. rosea cells after import of miR8788, miR1532, and miR1203 during interaction with wheat roots treated with these miRNA mimics