Isolation, Target Site Prediction and Validation of miR393 Prediction Target Gene Ve in Solanum torvum

In this study, total RNA was extracted by modified Triziol method and the full-length sequence of  Verticillium  wilt resistance gene  Ve  was obtained by homologous cloning. The binding site of miR393 to  Ve  was predicted by rna hybrid and validated by RLM-5’-RACE. The results showed that  Ve  gene contains 3302 nucleotides, encodes 904 amino acids, and has 85.63% homology with tomato, potato, tobacco, etc. The predicted results of binding sites showed that miR393 and  Ve  were almost completely paired from site 263 in  Solanum torvum . With the help of RLM-5’-RACE, we found that the miR393 and target gene  Ve  were cut between the 10 th  and 11 th  base sites of the complementary binding sequence. Through the verification of PCR and sequencing,  Ve  was finally confirmed as the target gene of miR393 in  Solanum torvum . It provides theoretical basis for further research on the molecular mechanism of miR393 mediated  Ve  gene participating in  Verticillium  wilt resistance.