Development of PCR Diagnosis of Pathogenic Fungi of the Genus Septoria, Affecting Cereal Crops in Northern Kazakhstan

I. S. Beyshova,¹ G. D. Chuzhebaeva¹, M. Zh Aubakirov¹,  A. S. Kozhmuhametova¹, A. A. Stakheev² , D. Y. Ryazantsev², S. K. Zavriev²,  A. T Oleynik³

¹Kostanay State University named after A. Baitursynov, 110000, Republic of Kazakhstan, Kostanai, st. A. Baytursynova 47 ²Institute of Bioorganic Chemistry of M. Shemyakin and Y.Ovchinniokov (Institute of Bioorganic Chemistry), Moscow. ³LLC "Kazakh Scientific and Research Institute of Plant Protection and Quarantine".

ABSTRACT: Traditional methods of complex diagnostics, including the selection of species in pure culture, selection environments and cultivation conditions, obtaining monoporosa isolates and microscopy are long, time-consuming and ineffective. Immune-enzyme analysis method takes a few hours, however, developed on the basis of the methodology of species-specific, predictive and quantification S. tritici and S. nodorum in the leaves and seeds of wheat insensitive do not always provide a clear identification of these species. The introduction of express methods of diagnostics of Septoria leaf blotch on the basis of the polymerase chain reaction (PCR) in modifications of Real-Time lets quickly and accurately diagnose the disease. A system of highly specific sensitive PCR diagnostics of pathogenic fungi Septoria tritici and Stagonospora nodorum, causing diseases of cereals in Northern Kazakhstan is developed. The primers and probes are designed based on partial sequences of the internal transcribed spacer of ribosomal DNA (ITS) provides a quick and accurate identification of pathogens Septoria by quantitative PCR without the risk of contamination of the work area with amplification products. Effectiveness of the test-system was also demonstrated in samples of total DNA isolated from the infected herbarium material.

KEYWORDS: genus Septoria; diagnostics; polymerase chain reaction; primer probe

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