A TuMV-NIb gene fragment was amplified from the total RNA of Mustard leaves infected with TuMV-GZ strain by RT-PCR (Reverse Transcription Polymerase Chain Reaction). This fragment was cloned into plant binary vector pBI121 in the antisense orientation and transformed into Mustard (Brassica juncea L. Linshicaoyaozi) by Agro bacterium tumefaciens mediated method. 11transgenic T0 plants were obtained. PCR (Polymerase Chain Reaction), ELISA (Enzyme-linked Immunosorbent Assay) and Southern blot analysis showed that the TuMV-Nib gene had integrated into the genome of mustard and the heredity of this gene in some lines followed the 3:1 ratio. Both T0 and T1 transgenic plants showed high levels of resistance to TuMV when artificially inoculated with TuMV. Northern blot analysis of T0 transgenic mustard revealed the integrated TuMV-NIb gene can be expressed normally.