Identification of Tilletia species using rep-PCR fingerprinting technique

Abstract

Analyzing 167 non-processed seed samples of wheat, it was found that 145 samples (86.8 %) were contaminated with Tilletia species, while 22 (13.2 %) samples were not contaminated. By using rep-PCR fingerprinting technique, it was found that DNA isolates of T. tritici originated from Serbian wheat samples had 80 % similarity with positive control for T. tritici. One isolate shared similarity of 60% with T. tritici, T. controversa and T. laevis. It was supposed that this isolate belongs to T. bromi. Isolate of T. laevis shared a similarity of 70 % with isolates of T. tritici and T. controversa, while T. walkeri was more than 10 % similar with T. tritici, T. controversa and T. laevis. Although T. controversa and T. tritici had high percent of genetic similarity, they were clustered separately. Our results suggest that rep-PCR fingerprinting could be a useful tool for monitoring presence of morphologically similar Tilletia species in wheat production areas

Development of the Droplet Digital PCR to Detect the Teliospores of Tilletia controversa Kühn in the Soil With Greatly Enhanced Sensitivity

“…Currently, molecular techniques to differentiate T. controversa from other similar Tilletia species are mainly focused on genetic diversity (Pimentel et al, 1998;Pimentel, 2000), PCR (Kochanová et al, 2004), repetitive extragenic palindromic PCR (Rep-PCR) (McDonald et al, 2000;Zupunski et al, 2011), primer-mediated asymmetric-PCR (RM-PCR) together with SYBR Green I and Taqman real-time PCR was also constructed with the detection limit of 0.1 fg and 1.0 fg, respectively (Yuan et al, 2009). Pieczul et al (2018) reported to identify Tilletia spp.…”

Development of ISSR-derived SCAR Marker and SYBR Green I Real-time PCR Method for Detection of Teliospores of Tilletia laevis Kühn

“…Vesna et al . 17 reported the differentiation of Tilletia species by repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting. However, many molecular markers have been developed to differentiate T .…”

Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR

“…Several studies have tried to identify specific markers for Tilletia species based on ITS, IGS1, and RPB2, but their results are not satisfactory 12 , 13 . However, rep-PCR fingerprinting, RAPD primer-mediated asymmetric PCR (RM-PCR), and sequence-characterized amplified region (SCAR) markers based on amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) have been employed to successfully distinguish T. controversa from its related species 14 – 18 . Hence, DNA marker technology may be a powerful tool to distinguish T. laevis from other related species, especially on quantification aspects.…”

“…sequence-characterized amplified region (SCAR) markers based on amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) have been employed to successfully distinguish T. controversa from its related species [14][15][16][17][18] . Hence, DNA marker technology may be a powerful tool to distinguish T. laevis from other related species, especially on quantification aspects.…”