| Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici(Pst), is one of the most important diseases threatening to global wheat production. At present, cultivation of disease-resistant varieties is the most effective, economic and environment-friendly method to control wheat stripe rust. However, most of named resistance genes from wheat have been overcomed by new virulent Pst races, and conventional cross-breeding often lags behind the variation of wheat stripe rust.Therefore, original strategies must be changed, and new rust resistanceresource must be explored to ensure food safety in production and achieve durable disease control of wheat stripe rust in our country.Cellular signal transduction in organisms include molecular pathways of cell sensation and transduction environmental stimuli, the expression of regulated genes, and regulation of physiological responses. Plant-pathogen interaction involves a series of processes of signal recognition, signal transduction and defense response activation. The growth, development and pathogenicity of pathogenic fungi are also dependent on regulation of extracellular signal transduction pathways. Previous studies showed that protein kinase play an important role in the growth of fungal hyphae, virulence, pathogenicity.To obtain the predicted important protein kinases in Pst,we search homologous sequences of the important or essential kinases from Fusarium graminearum from the Pst genomic protein database. To obtain the important protein kinases, we perform the quantitative real-time pcr(q RT-PCR) analysis and virus induced the Pst gene silencing(VIGS) analysis. To obatin the stable transgenic stripe rust resistant materials, we introduce the inverted repeat sequences of important kinases into the wheat using the particle bombardment method according to the principle of host induced gene silencing(HIGS).1. The 27 predicted important protein kinases were obtained by searching homologous sequences.2. The gene expression profile analysis of predicted important protein kinases during the Pst various asexual development stages were completed by using q RT-PCR. The genes were induced in the infection process including Ps23940, Ps18181, Ps19498, Ps19627, Ps9318, Ps23739 and Ps26471; Ps1134, Ps25317, Ps9666, Ps20225, Ps21460 and Ps26267 were highly expressed in the early infection process(0h–48h); Ps27645, Ps16944, Ps20880 and Ps23097 were specific highly expressed at the stage of resting urediospores; The amount of transcripts of Ps19627, Ps9318, Ps21460, Ps23097 and Ps23739 were increased larger at the spore production stage than that at the colony extension stage.4. Knocking down Ps12169, Ps16944, Ps18693, Ps23739, Ps9666, Ps26267, Ps18181, and Ps11901 by VIGS showed that transient silencing of these protein kinase genes would decrease the number of urediosorus of Pst significantly. The result was that all gene expression were silenced effectively exclude Ps12169 using q RT-PCR analysis. These results suggest that Ps16944, Ps18693, Ps23739, Ps9666, Ps26267, Ps18181 and Ps11901 might play an important role in regulating Pst development and pathogenicity.5. The two RNAi vectors, p AHC25-Ps26267 and p AHC25-Ps23739, were introduced into the wheat of Xinong1376 by the particle bombardment method. Now, we have acquired T1 transgenic wheat. |
