Plant Science & Physiology

Biography

Biography: Anna Kot

Abstract

The species Puccinia triticina (Pt) and Puccinia striiformis f. sp. tritici (Pst) are devastating cereal pathogens that cause brown/leaf and yellow/stripe rust diseases, respectively. Both fungi are an obligate parasites capable of producing infectious urediniospores as long as infected leaf tissue remains alive. They are responsible for significant yield losses of common wheat (Triticum aestivum L.) that may have ranged from 10% to 70% depending on susceptibility of the cultivar, initial infection rate, development and duration of disease. The early detection of fungal pathogens can lead to preventive measures and minimize economic losses through i.e. fungicidal control. The molecular methods are the most reliable for monitoring of disease development and early pathogens detection (Fig 1). Multiplex PCR has the potential to target and differentiate more than one species at the same time.

In the present study, we develop a conventional PCR assays for the simultaneous species-specific detection of Pt and Pst. The Multiplex PCR assay targets the second largest subunits of the RNA polymerase II (rpb2) for Pt and beta-tubulin 1 (tub1) genes for Pst. The specificities of the PCR primers were verified using naturally infected plant materials with visual symptoms of rust diseases and diseases caused by other whet pathogens (Blumeria graminis, Drechslera tritici-repentis and Septoria tritici). The primer sets LidPs9/10 and LidPr1/2 produced a single DNA fragments of lengths 240 and 144 bp, respectively. No cross-reactions were observed with tested fungal pathogens and healthy plant tissues. The detection limit of singular primer sets was reached at 1 pg for Pt and 50 pg for Pts. The assay shows 100% effectiveness in rust fungi detection that make them promising tools for determining the proper schedule for plant protection.