Study On The Transgenic Tobacco Expressing Protein Kinase Like Gene TA50-10 Of Wheat And Analysis Of Disease Resistance

TA50-10 is a truncated gene cloned from wheat cDNA. It encodes a Protein Kinase like domain. The prokaryotically expressed TA50-10 could significantly induce local resistance against Tobacco mosaic virus (TMV) infection when applied by spraying onto the tobacco leaves.In this study, we excised the TA50-10 gene by digestion with restriction enzymes BamH I and XbaI, and inserted it into pBI121 to make the recombinant construct pB-50-10. pB-50-10 was transformed into Agrobacterium tumefaciens EHA105 by the freeze-thaw method.Tobacco transformation was conducted by infecting tobacco leaf discs with recombinant A. tumefaciens strain EHA105 containing pB-50-10. The transgenic tobacco plants were obtained under selection of kanamycin resistance. After PCR analysis,38 To plants were identified as transgenic tobacco. GUS staining analysis showed that the TA50-10 expressed differently at the protein level in the transgenic tobacco. RT-PCR analysis showed that the TA50-10 expressed at the transcriptional level in the transgenic tobacco.The transgenic tobacco showed significant resistance against TMV infection and suppressed the multiplication of TMV. The activity of POD and PPO in the transgenic plants increased than the non-transgenic control. The transgenic tobacco plants also have anti-invasive ability against Pectobactrium carotovorum subsp. carotovorum (Pcc) and Ralstonia solanacearum E.F.Smith.As TA50-10 was cloned from safe food crop wheat, it is considered to be of great value in disease resistance breeding for tobacco NC89.