At 10 days

after the virus inoculation, the BSMV CP transcript was detected in plants inoculated with BSMV virus, but not in the mock plants, revealing successful virus infection. As expected, the TaWAK5 transcript levels were considerably reduced in CI12633 plants infected by BSMV:TaWAK5 ( Fig. 6-A), suggesting that the TaWAK5 transcripts were silenced in these CI12633 plants infected Thiazovivin datasheet by BSMV:TaWAK5. In disease screening tests of non-infected plants, the 4th sheaths of mock-treated CI12633 and those infected with the BSMV:TaWAK5 virus were inoculated with mycelia of R. cerealis. The 4th sheaths of the susceptible cultivar Wenmai 6 were used as a positive control to show successful R. cerealis inoculation. At 2 weeks post R. cerealis inoculation, lesions with dark-brown margins (an early symptom of sharp eyespot disease) were observed on the 4th sheaths of the susceptible Wenmai 6, but not on the BSMV:TaWAK5-inoculated, BSMV:GFP-inoculated, or mock-treated CI12633 plants ( Fig. 6-B). The resistance continued to be present through more mature stages. No sharp eyespot symptoms were observed at 4th sheaths and stems of BSMV:TaWAK5-inoculated, BSMV:GFP-inoculated, and mock CI12633 plants, but obvious symptoms were present on the 4th sheaths and stems of Wenmai 6 plants. These results suggested that the silencing of TaWAK5 did not directly compromise wheat

resistance to R. cerealis in CI12633. In this study, we isolated a novel wheat WAK gene, TaWAK5, from R. cerealis-resistant wheat CI12633, based on a cDNA transcript that was differentially GSK2118436 mouse expressed between resistant wheat genotype CI12633 and susceptible wheat cultivar Wenmai 6. PDK4 qRT-PCR analysis revealed that the

transcript abundance of TaWAK5 in wheat was rapidly increased by R. cerealis infection. Additionally, TaWAK5 in the R. cerealis-resistant lines was induced to higher levels than in R. cerealis-susceptible lines at 7 dpi with R. cerealis. These results suggested that TaWAK5 may be involved in wheat defense responses to R. cerealis infection. Sequence analysis and phylogenetic analysis revealed that TaWAK5 was a member of the WAK sub-group of the RLK family in wheat. Several WAK genes have been shown to play important roles in regulating plant defense responses. WAK1 from Arabidopsis and OsWAK1 from rice were shown to enhance resistance to the pathogens B. cinerea and M. oryzae, respectively [5] and [11]. TaWAK5 is a non-RD-type protein, as it has an HGD motif in its subdomain VIb. Out of 38 receptor kinases tested in plants, the six which fall into the non-RD class all function in disease resistance and act as PRRs, while the remaining 32 kinases are RD or alternative catalytic function (ACF) kinases and are involved in developmental processes [35], suggesting that all the non-RD RLKs seemed to participate in innate immunity.