The development of a PCR-based method for detecting Puccinia striiformis latent infections in wheat leaves

Abstract

Stripe rust of wheat caused by Puccinia striiformis f. sp. tritici is one of the most important diseases on wheat worldwide, especially in temperate regions with cool moist weather conditions. A rapid and reliable detection of the pathogen in latent infected wheat leaves during overwintering of the fungus in the dormant stage will contribute to determine the initial inoculum potential and thus to predict early outbreak and to improve effective management of the disease. To achieve this aim, a PCR-based method was developed for specific and sensitive detection of P. striiformis. Specific primers were designed according to a genome-specific sequence of P. striiformis. To evaluate the specificity of the primers, seven different isolates and races of P. striiformis as well as six other pathogens of wheat were tested. All isolates of P. striiformis yielded a distinct band of a fragment of 470 bp, while using DNA of the other wheat pathogens as a template no amplification product was detected. The sensitivity of the primers was tested using serial dilutions of total DNA from P. striiformis; the limit of detection was 10 pg of DNA. Using extracts from P. striiformis-infected wheat leaves, the fungus could be determined in the leaves before symptoms appeared. The stripe rust could also be detected in the dormant stage by the PCR assay in samples of wheat leaves taken during the winter season. The application of the PCR assay may be useful for rapid and reliable detection of P. striiformis in latent infected leaves of overwintering wheat plants.

A Review of Conventional PCR Assays for the Detection of Selected Phytopathogens of Wheat

“…The reliability of that assay was confirmed under greenhouse and field conditions with naturally infected leaves even before any visible symptoms. Wang et al [2008] developed a specific and sensitive detection method based on the P. striiformis repeat (PSR) sequence suggested by Zheng et al [2000a, b] as a useful target for specific detection by PCR. The specificity of PST2 f/r primers was tested and confirmed against healthy plant tissue, P. striiformis , and other 6 isolates of fungi.…”

“…There are many detection systems for F. sporotrichioides , P. striiformis f. sp. tritici , and Z. tritici [Beck and Ligon, 1995;Consolo et al, 2009;Demeke et al, 2005;Fraaije et al, 1999Fraaije et al, , 2001Gao et al, 2016;Guo et al, 2006;Konstantinova and Yli-Mattila, 2004;Lihua et al, 2008;Niessen et al, 2004;Wang et al, 2008Wang et al, , 2009Wilson et al, 2004;Zhao et al, 2007]. Particularly for Z. tritici , the number of known diagnostic tests is justified by the significance of STB, one of the most important foliar diseases of wheat caused by this fungus [Keon et al, 2007;Shetty et al, 2007].…”

- Strategies of Nitrosomonas europaea 19718 to Counter Low Dissolved Oxygen and High Nitrite Concentrations

“…Origin confirmation for genes homologous to the stem rust pathogen Genomic DNA was extracted from the urediniospores of CYR23 and leaves of Suwon 11 according to the protocol described by Wang et al [66]. Specific primers for possible Pst genes were designed using the software of Primer Premier 5.0.…”

Role of Resistance (Yrs) Genes to Puccinia Striiformis in Five Bread Wheat Cultivars Susceptible and Validation by Molecular Markers in Assessment of Resistance

“…(McIntosh et al 2005). Molecular markers have been linked to many yellow rust resistance genes in wheat, such as Yr5, Yr10, Yr15, Yr17, Yr24, Yr26, Yr29, Yr32, Yr34 and YrH52 (Chague et al 1999;Robert et al 2000;Peng et al 2000;Sun et al 2002;and Wang et al 2008). These markers provide an important tool to plant breeders for marker-aided wheat breeding and also for pyramiding resistance genes in the absence of distinguishable rust virulence's (Kaur et al 2008).…”