AFLP-based PCR markers that differentiate spot and net forms of Pyrenophora teres

Abstract

Specific polymerase chain reaction (PCR) primers were developed from amplified fragment length polymorphism (AFLP) fragments of Pyrenophora teres, the causal agent of net blotch on barley leaves. The primers were designed specifically to amplify DNA from P. teres f. teres (net form) and allow its differentiation from P. teres f. maculata (spot form), which is morphologically very similar to P. teres f. teres in culture. The PCR amplification was carried out successfully from DNA extracted from fungal mycelium. The PCR assay was validated with 60 samples of Pyrenophora species. The amplification with four designed PCR primer pairs provided P. teres form-specific products. No cross-reaction was observed with DNA of several other species, such as P. tritici-repentis, P. graminea and Helminthosporium sativum.