| Wheat stripe rust, caused by a pathogen Puccinia striiformis f. sp. tritici(Pst), is an important air-borned fungal disease and occurs almost in wheat production all above the world. Meanwhile, this diease happens in large scale area and lead to serious loss of wheat with suitable climate annual, and may become the key factors affecting the sustainable development of wheat production in China. As everyone knows that the wheat stripe rust is obligate parasitic fungi which absorbs nutrients by haustoria, but the transcriptome and its function of stripe rust haustoria are rarely analysed for a long time, limits the further system research of the disease prevention and control technology. At the transcriptome level,therefore, a comprehensive system to understand the gene expression characteristics of wheat stripe rust was interactions, cloning article anti rust related genes and use virus induced gene silencing technology analysis of its function, to illustrate the molecular mechanism of wheat and the rust interactions.Therefore, this thesis proposed in previous article rust based on genome sequencing, the purpose of this experiment is intended to analyze wheat rust infection, we measured the wheat stripe rust haustoria transcriptome, predict the candidate effect protein(effector candidates),and the function of PSTha5a23 was studied.The main results were as follows:1. This experiment by using the rust CYR32 infected wheat leaf homogenate after 20 μm nylon membrane filtration and separation of haustoria, haustoria were isolated woth the purity of 62%, the haustoria RNA concentration was 83 ng/μL.2. 13211 genes were got by the Seolxa RNA-seq sequencing technology of haustoria,among them, 1547 specific haustoria genes were found.3. The unigene GO annotation classification results show that a total of 883 unigene annotations to biological processes(in process), cell components(cellular component)and molecular functions( molecular function) 3 classes. Among of them, 293 unigene in biological processes, 85 unigene in cell components, and 491 unigene in molecular function.4. The unigene sequences is mapped to the KEGG database reference metabolic pathways(pathway), a total of 1489 unigene commented, involving 20 metabolic pathways,including 16 no-repeat metabolic pathways.5. 51 new genes get involved in the transmembrane transport analysied by SignalP and TMHMM. 49 new genes get involved in the anchor and modified protein analysied by PredGPI.6. Here, we report our findings on an in planta highly induced candidate effector from Pst, PSTha5a23. The PSTha5a23 gene is unique to Pst and shows a low level of intra-species polymorphism. It has a functional N-terminal signal peptide and is localized to the wheat cytoplasm. Overexpression of PSTha5a23 in Nicotiana benthamiana was found to suppress programmed cell death triggered by BAX, PAMP-INF1, and two resistance-related mitogen-activated protein kinases(MKK1 and NPK1). Overexpression of PSTha5a23 in wheat also suppressed pattern-triggered immunity(PTI)-associated callose deposition. In addition, silencing of PSTha5a23 did not change Pst virulence phenotypes; however,overexpression of PSTha5a23 significantly enhanced Pst virulence in wheat. These results indicate that the Pst candidate effector PSTha5a23 plays an important role in plant defense suppression and rust pathogenicity, and also highlight the utility of gene overexpression in plants as a tool for studying effectors from obligate biotrophic pathogens. |
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