Detection of Tilletia controversa and Tilletia caries in wheat by PCR method

Kochanová, M.; Zouhar, Miloslav; Prokinová, E.; Ryšánek, P.

doi:10.17221/3684-pse

Cited by 22 publications

(15 citation statements)

References 9 publications

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“…The toolkit of DNA sequence based assays for the regulated species is as yet limited to one or a few options and gene regions, especially for DB (Liu et al 2009;Zouhar et al 2010;Gao et al 2014) and CB (Josefsen and Christiansen 2002;Kochanová et al 2004) and studies demonstrating their efficacy and specificity during regulatory inspection of field samples are lacking. Thus, despite the known limitations inherent in morphology-based identifications, federal diagnostic labs continue to rely on labour-and timeintensive seed-washing and microscopy techniques (IPPC 2016) to screen grain importations for teliospores of regulated Tilletia species, at least in Canada.…”

Section: Introductionmentioning

confidence: 99%

Genome sequencing and comparison of five Tilletia species to identify candidate genes for the detection of regulated species infecting wheat

et al. 2019

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Tilletia species cause diseases on grass hosts with some causing bunt diseases on wheat (Triticum). Two of the four species infecting wheat have restricted distributions globally and are subject to quarantine regulations to prevent their spread to new areas. Tilletia indica causes Karnal bunt and is regulated by many countries while the nonregulated T. walkeri is morphologically similar and very closely related phylogenetically, but infects ryegrass (Lolium) and not wheat. Tilletia controversa causes dwarf bunt of wheat (DB) and is also regulated by some countries, while the closely related but non-regulated species, T. caries and T. laevis, both cause common bunt of wheat (CB). Historically, diagnostic methods have relied on cryptic morphology to differentiate these species in subsamples from grain shipments. Of the DNA-based methods published so far, most have focused on sequence variation among tested strains at a single gene locus. To facilitate the development of additional molecular assays for diagnostics, we generated whole genome data for multiple strains of the two regulated wheat pathogens and their closest relatives. Depending on the species, the genomes were assembled into 907 to 4633 scaffolds ranging from 24 Mb to 30 Mb with 7842 to 9952 gene models predicted. Phylogenomic analyses confirmed the placement of Tilletia in the Exobasidiomycetes and showed that T. indica and T. walkeri were in one clade whereas T. controversa, T. caries and T. laevis grouped in a separate clade. Single copy and species-specific genes were identified by orthologous group analysis. Unique species-specific genes were identified and evaluated as suitable markers to differentiate the quarantine and non-quarantine species. After further analyses and manual inspection, primers and probes for the optimum candidate genes were designed and tested in silico, for validation in future wet-lab studies.

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Section: Introductionmentioning

confidence: 99%

Genome sequencing and comparison of five Tilletia species to identify candidate genes for the detection of regulated species infecting wheat

et al. 2019

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“…For example, Tilletia caries (syn. T. tritici) and T. controversa (causing common and dwarf bunt of wheat, respectively) have been successfully detected or quantified on seeds and in the plant by PCR (Josefsen and Christiansen 2002;Eibel et al 2005a;Kochanová et al 2004;Kellerer et al 2006;Roberts et al 2007;Yuan et al 2009;Zouhar et al 2010). Qualitative PCR was used by Schenck (1998) and Singh et al (2004) to diagnose systemic infections of Ustilago scitaminea in sugarcane and by Willits and Sherwood (1999) and Shen et al (2006) who studied the spread of Ustilago hordei and Sporisorium reiliana, respectively, in differentially resistant barley and maize varieties.…”

Section: Introductionmentioning

confidence: 99%

Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR

Wunderle¹,

Leclerque²,

Schaffrath

et al. 2012

Eur J Plant Pathol

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Loose smut fungi of barley and wheat (Ustilago nuda and U. tritici, respectively) colonize the plant without causing obvious disease symptoms before heading. The availability of diagnostic methods to detect and follow the growth of these pathogens in the plant would therefore be highly advantageous for both resistance breeding and the development of effective seed treatments. Using seed lots of barley and wheat highly infected with loose smut, we studied the early establishment of the loose smut pathogens in the plant by fluorescence microscopy. In hand-cut sections stained with the fluorochrome Blankophor®, fungal hyphae were observed to invade the shoot apical meristem and leaf primordia during the first days after the onset of germination. At the first node stage the ear and leaf primordia were generally extensively colonized. Hyphae of U. nuda were also regularly observed in high density in the nodes. A protocol was developed for the specific amplification of U. nuda and U. tritici DNA extracted from infected plant tissue. PCR screening of U nuda in seedlings from infected and healthy seed lots was compared to ELISA, microscopy and ultimately head infection of mature plants derived from tillers of the tested seedlings. The results indicated that a prediction of loose smut infection by real-time PCR is possible at the second leaf stage, and that the assay is equally suited for use with spring and winter varieties of barley and wheat.

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“…Currently, the disease is registered in many countries in the area between 41 и 530 north latitude in Europe, for example, in the Czech Republic [6], and between 37 and 380 north latitude in North America. Field researches conducted by B. Goates, G.L.…”

Section: Expansion and Harmfulness Of The Wheat Dwarf Smutmentioning

confidence: 99%

Biodiversity of Plant Pathogens in the Context of International Economic Issue: Case Study of Tilletia Controversa Kuehn

Shutko

2018

International Scientific and Practical Conference "AgroSMART - Smart Solutions for Agriculture" (AgroSMART 2018)

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Globalization as a process of world economic, political and cultural integration requires strengthening of control over dangerous plant diseases, pests and weeds. This is evidenced by strengthening of phytosanitary requirements in some countries (the People's Republic of China imposed a ban on wheat supplies from the European part of the Russian Federation due to a quarantine object-dwarf smut Tilletia controversa Kuehn found in wheat; Jordan reduced the permissible Fusarium infection rate of wheat grain to 0,03%), and environmental disasters caused by invasive species

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Detection of Tilletia controversa and Tilletia caries in wheat by PCR method

Cited by 22 publications

References 9 publications

Genome sequencing and comparison of five Tilletia species to identify candidate genes for the detection of regulated species infecting wheat

Genome sequencing and comparison of five Tilletia species to identify candidate genes for the detection of regulated species infecting wheat

Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR

Biodiversity of Plant Pathogens in the Context of International Economic Issue: Case Study of Tilletia Controversa Kuehn

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