Study On Signal Pathway Of N’ Gene In Nicotoana Sylvestris Resist To TMV

In order to explore signaling pathway associated with R-gene N’in Nicoiiana sylvestris resistance to TMV. Eight genes of signal components, PAD4, EDS1, Rar1, SGT1, NPR1, Jar6, COI1,CTR1, were transformed into the plant, and symptom were evaluated. The main results are as follows:1. Total RNA of N. sylvestris was extracted with the method of Trizol, and eight genes were cloned using RT-PCR.2. The reverse complementary sequences of the eight genes were ligased into plant expression vector pCAM2300-35s-OCS after digested with enzyme, ligased and transformed. Eight RNAi vectors were established.3. The RNAi vectors were infected with Agrobacterium tumefaciens LBA4404. All above genes except Rarl were transfered into N. sylvestris with the method of leaf disc transformation.4.Virus strain TMV-T1was inoculated on some leaves of the transgenetic plants. Only on the plants Jar6transformed, symptoms of necrosis were exhibited on inoculated leaves, and mosaic were expressed on no-inoculated leaves, which different from on wild variety of N. sylvestris. In contrast, necrosis exhibited both other transgenic plants and their wild variety. This implies TMV was not limit within inoculated area of the transgenic plants with Jar6.