| Barley leaf strip, caused by the seed-borne fungus Pyrenophora graminea (Ito andKuribayashi)[anamorph Drechslera graminea (Rabenh. ex. Schlech.) Shoemaker], is one ofbarley diseases, The disease results in severe yield reduction, and it s difficult to prevent. Sobreeding resistant cultivars has become the most effective way to control barley leaf strip. Butthe research fall behind overseas in terms of resistance and quality of barley leaf strip, because ofthe lack of germplasm resources and correlational research of barley in China. Meanwhile,proteomics, mainly including2-dimensional gel electrophoresis (2-DE) and MALDI-TOF-MStechniques, provides a important approach. In this study, on the one hand, genetic diversity of20isolation from different areas in Gansu were analyzed by using30RAPD primers and resistantevaluation of barley varieties were identified by artificial innoculation; on the other hand,according to the results of pathogenicity test of strain and resistant evaluaiton of barley varieties,we chose Ganpi6(resistance) and Isotta (susceptible) as the experimental material, pathogen isisolate QWC, and studied the different resistance-related proteins to barley leaf strip withproteomic method. We studied the different resistance-related proteins to barley leaf strip afterartificial inoculation through infection Ganpi6(resistance) and Isotta (susceptible) by isolateQWC with proteomic method. The results were as follows：1. The RAPD result showed that30polymorphism primers and20isolates were analyzedby RAPD and classed into four main clusters based on RAPD date using UPGMA when geneticsimilarity is0.7166, isolate HYH and YC are respectively formed a single lineage, isolate JT andXTB are formed a single category, and others are formed a separate category. Isolate SW and QQ;Isolate QWC and HZ have the highest genetic similarity, and GS is0.9365; isolate YC isdifferent from others, and GS is0.6241. Thus there were Genetic difference among all isolates.2. The pathogenicity test indicated that14isolates can cause disease, but significantdifferences also existed among different isolates. The most virulence isolate was QWC, infectedrate is73.53%.3. The resistant evaluation of30barley varieties showed that visible differences existedamong different barley varieties. Of30varieties, no highly resistant variety had been found,5 were moderately resistant,19were susceptible and6were highly susceptible.5moderatelyresistant can be used disease-resistant parents, the resistance of Ganpi6is the best and Isotta isthe worst.4. Proteomics showed that5specific-expressed proteins were screened in Ganpi6inducedby isolate QWC at7days;11specific-expressed proteins were screened in Ganpi6at21days;4specific-expressed proteins were screened in Isotta induced by isolate QWC at7days;10specific-expressed proteins were screened in Isotta at21days. And these specific-expressedproteins were of significant difference of protein spot according to the T-test (p <0.05).5. We got these specific-expressed protein mass fingerprintings (PMFs) and their aminoacid sequences by MALDI-TOF-MS and MALDI-TOF-TOF MS/MS. By searching the relatedplant protein database using the MASCOT search engine, we found these proteins were relatedto disease resistance, protein biosynthesis, photosynthesis, energy metabolism, cellular signaltransduction etc in Ganpi6. Lipoxygenase2is the only defense protein. These proteins wererelated to disease resistance, photosynthesis, energy metabolism, respiratory action, cellulosebiosynthesis etc in Isotta, and defense protein is lectin.Based on the experimental results, we explored the molecular mechanism of barley leafstripe, elucidated barley resistance mechanisms to barley leaf stripe using proteomics. Theseresults will provide gene resources and theoretical principles for barley molecular resistancebreeding to Drechslera graminea, and it will be of great importance in excavating diseaseresistance genes and breeding resistant cultivars. |
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