Abstract
Clubroot, caused by Plasmodiophora brassicae, is one of the most detrimental threats to crucifers worldwide and has emerged as an important disease of canola (Brassica napus) in Canada. At present, pathotypes are distinguished phenotypically by their virulence patterns on host differential sets, including the systems of Williams, Somé et al., the European Clubroot Differential set, and most recently the Canadian Clubroot Differential set and the Sinitic Clubroot Differential set. Although these are frequently used because of their simplicity of application, they are time-consuming, labor-intensive, and can lack sensitivity. Early, preventative pathotype detection is imperative to maximize productivity and promote sustainable crop production. The decreased turnaround time and increased sensitivity and specificity of genotypic pathotyping will be valuable for the development of integrated clubroot management plans, and interest in molecular techniques to complement phenotypic methods is increasing. This review provides a synopsis of current and future molecular pathotyping platforms for P. brassicae and aims to provide information on techniques that may be most suitable for the development of rapid, reliable, and cost-effective pathotyping assays.
Highlights
Clubroot DiseaseClubroot, caused by the obligate parasite Plasmodiophora brassicae Woronin, is one of the most serious soilborne diseases of crucifers worldwide
In comparison with the previously mentioned single nucleotide polymorphisms (SNPs)-based PCRs, where amplification proceeds based on the existence of a specific allele, SNaPshot allows for the detection of up to four allelic variants as bases are distinguished by means of fluorescent dideoxy nucleotide triphosphates (ddNTPs)
In addition to basic PCR reagents, a clean-up kit is needed to purify PCR products, SNaPshot reagents are necessary for the extension reaction, shrimp alkaline phosphatase or calf intestinal phosphatase is needed for post-extension treatment, Hi-Di Formamide is required as an injection solvent in the genetic analyzer, and a size standard is required to investigate the results of the fluorescent peaks
Summary
Clubroot, caused by the obligate parasite Plasmodiophora brassicae Woronin, is one of the most serious soilborne diseases of crucifers worldwide. As the number of clubroot-infested fields began to increase, P. brassicae was declared a pest under the Agricultural Pests Act of Alberta in April of 2007 [7]; this designation enabled the enforcement of control measures throughout the province aimed at reducing the dissemination and impact of the disease. Clubroot is endemic to much of central Alberta [1], with more than 3000 confirmed field infestations [8] and the disease having been identified in 44 of the 66 counties or municipal districts where canola is grown [9]. Epidemiological models had predicted that clubroot could spread beyond Alberta throughout the Canadian Prairies, and since the disease has been confirmed on canola in Saskatchewan and Manitoba [3]. Clubroot occurs in canola crops in Ontario, Canada [10], and in North Dakota, USA [11]
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