QTL mapping and proteomic profiling of barley: insights into resistance and susceptibility to Pyrenophora teres f. teres

Pyrenophora teres f. teres (Ptt), the causal agent of net form net blotch disease in barley, is an economically important fungal pathogen worldwide. Understanding both host resistance mechanisms and pathogen virulence factors is essential for developing durable net form net blotch resistant barley cultivars. Quantitative trait loci (QTL) mapping was conducted using a cross between two Ptt isolates, one virulent on the barley cultivar Prior and the other being avirulent. A major QTL associated with virulence on Prior was detected on chromosome 5. A progeny isolate possessing this QTL, together with the two parental isolates, was subsequently used in the proteomic analyses. Label-free proteomics was used to quantify in planta the protein profile changes in Prior following inoculations with the virulent and avirulent parental Ptt isolates, and the virulent progeny isolate. Leaf samples were collected at two (D2) and five (D5) days post-inoculation, and proteomic analyses performed to identify proteins associated with host resistance and pathogen virulence. A dataset comprising 2,886 barley proteins and 51 Ptt proteins was analysed. Principal component analysis (PCA) of the barley Prior proteomes revealed distinct clustering based on resistance and susceptibility at D5, while D2 samples formed a separate cluster. The PCA of the Ptt proteomes identified separate clusters, one comprised of the D2 and D5 avirulent parental isolate and another cluster of the virulent isolates at D5 only. Gene ontology analysis of the Prior proteins that were significantly increased in the resistant compared to the susceptible groups revealed functional categories related to protein translation, biosynthesis and chloroplast activities. The proteins that were significantly increased in the susceptible compared to the resistant Prior group were associated with organic acid and carbohydrate metabolism. The proteomic profiles and bioinformatic analysis generated in our study provide novel insights into the molecular basis of resistance and virulence in the barley-P. teres pathosystem. Key messageThis study reveals the first in planta proteomic profiles of both barley and Pyrenophora teres f. teres, identifying unique virulence-associated proteins and host responses linked to resistance and susceptibility.