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. 2021 Mar 25;186(2):853–864. doi: 10.1093/plphys/kiab146

Figure 3.

Figure 3

TRSV-based silencing of miR166 in different cucurbits plants. A, Schematic representation of VbMS of mi166. “–-” represents no nucleotide in this position. The STTM166 sequence was introduced into pTRSV2 downstream of the CP stop codon. The 48 nucleotides stem-loop spacer contributes to STTM stability and efficiency. B, Leaf phenotypes of plants inoculated with TRSV-STTM166 at 23 dpi. The arrows indicate the ectopic leaf outgrowing from the vein. Scale bars = 1 cm in the third column. The fourth column shows a magnification of dotted rectangle regions in the third column. C, Stem-loop RT-qPCR detection of miR166 level in TRSV- and TRSV-STTM166-infected cucurbits plants. D, Relative expression level of miR166 target gene in various cucurbits plants inoculated with TRSV or TRSV-STTM166. In (C) and (D), the biological experiment was repeated three times. Error bars represent ± SD. Statistical significance between treatments was determined using paired Student’s t test: *P < 0.05, **P < 0.01, ***P < 0.001. E, RT-PCR detection confirmed the TRSV infection as indicated by the presence of CP transcripts in all plants and the expression of STTM166