Use of Specific Repetitive Sequences in<i>Peronospora tabacina</i>for the Early Detection of the Tobacco Blue Mold Pathogen

Wiglesworth, M. D.; Nesmith, William C.; Schardl, Christopher L.; Li, Daoxin; Siegel, Malcolm R.

doi:10.1094/phyto-84-425

Cited by 28 publications

(22 citation statements)

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“…Alternative strategies were developed for the design of species or strain-specific markers using randomly selected sequences (Wiglesworth et al, 1994;Boehm et al, 2001). Sequence Characterized Amplified Regions (SCAR) can be selected from RAPD (Random Amplified Polymorphic DNA) fingerprints and used to mark specific alleles (Paran and Michelmore, 1993) or generate speciesspecific PCR markers (Schilling et al, 1996;Schubert et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

SCAR–based PCR primers to detect the hybrid pathogen Phytophthora alni and its subspecies causing alder disease in Europe

et al. 2005

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Section: Introductionmentioning

confidence: 99%

SCAR–based PCR primers to detect the hybrid pathogen Phytophthora alni and its subspecies causing alder disease in Europe

et al. 2005

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“…The technique was successfully established for further plant pathogens in the following years (Wiglesworth et al 1994;Goodwin et al 1995;Sreenivasaprasad et al 1996;Tooley et al 1997;Lindqvist et al 1998;Bakan et al 2002;Casimiro et al 2004). PCR-based diagnosis of tobacco blue mold infection has already been described in 1994 (Wiglesworth et al 1994) and has been renewed recently using ITS DNA (Ristaino et al 2007). The aim of these studies was either to detect early infections (Caiazzo et al 2006) or differentiation from other tobacco pathogens (Ristaino et al 2007).…”

Section: Discussionmentioning

confidence: 99%

“…1b). The sonication as described by Wiglesworth et al (1994) turned out to be an appropriate method for DNA extraction from minute sample amounts, despite the risk of DNA fragmentation as discussed by Caiazzo et al (2006).…”

Section: Phenotype-specific Primersmentioning

confidence: 99%

PCR-based monitoring of recent isolates of tobacco blue mold from Europe reveals the presence of two genetically distinct phenotypes differing in fungicide sensitivity

2008

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Bioassays testing the fungicide sensitivity against metalaxyl of Peronospora tabacina isolates collected in German tobacco fields in 2005 revealed the presence of two phenotypes, resistant and sensitive. DNA fingerprints using SSR and minisatellite primers allowed separation of the samples into two groups. The differences in amplification patterns coincided with the sensitive and resistant reaction of the isolates in metalaxyl bioassays. New primers were developed which allowed PCR-based detection of P. tabacina and differentiation of the metalaxyl-sensitive and the metalaxyl-resistant phenotype, respectively.

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“…Although some authors have reported other multicopy genes, including the mitochondrial cytochrome oxidase ( cox ) gene and the Ypt1 gene as alternatives to ITS regions, in some cases, it is difficult to distinguish hybrid species and closely related Phytophthora species e.g., P. cactorum / P. idaei , and P. iliciss / P. nemerosa [11, 12]. An alternative approach would be to develop specific primers using random amplified polymorphic DNA (RAPD) since RAPD can be applied for the design of species or strain-specific markers from randomly amplified sequences [13, 14]. Thus, sequence characterized amplified region (SCAR) markers specific to fungal pathogens can be derived from the sequence of a specific strain or gene, or from sequences randomly amplified by decamers [15].…”

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confidence: 99%

Development of SCAR Markers for the Identification ofPhytophthora katsuraeCausing Chestnut Ink Disease in Korea

Lee

et al. 2013

Mycobiology

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Sequence characterized amplified region (SCAR) markers are one of the most effective and accurate tools for microbial identification. In this study, we applied SCAR markers for the rapid and accurate detection of Phytophthora katsurae, the casual agent of chestnut ink disease in Korea. In this study, we developed seven SCAR markers specific to P. katsurae using random amplified polymorphic DNA (RAPD), and assessed the potential of the SCAR markers to serve as tools for identifying P. katsurae. Seven primer pairs (SOPC 1F/SOPC 1R, SOPC 1-1F/SOPC 1-1R, SOPC 3F/SOPC 3R, SOPC 4F/SOPC 4R, SOPC 4F/SOPC 4-1R, SOPD 9F/SOPD 9R, and SOPD 10F/SOPD 10R) from a sequence derived from RAPD fragments were designed for the analysis of the SCAR markers. To evaluate the specificity and sensitivity of the SCAR markers, the genomic DNA of P. katsurae was serially diluted 10-fold to final concentrations from 1 mg/mL to 1 pg/mL. The limit of detection using the SCAR markers ranged from 100 µg/mL to 100 ng/mL. To identify the limit for detecting P. katsurae zoospores, each suspension of zoospores was serially diluted 10-fold to final concentrations from 10 × 105 to 10 × 101 zoospores/mL, and then extracted. The limit of detection by SCAR markers was approximately 10 × 101 zoospores/mL. PCR detection with SCAR markers was specific for P. katsurae, and did not produce any P. katsurae-specific PCR amplicons from 16 other Phytophthora species used as controls. This study shows that SCAR markers are a useful tool for the rapid and effective detection of P. katsurae.

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Use of Specific Repetitive Sequences inPeronospora tabacinafor the Early Detection of the Tobacco Blue Mold Pathogen

Cited by 28 publications

References 0 publications

SCAR–based PCR primers to detect the hybrid pathogen Phytophthora alni and its subspecies causing alder disease in Europe

SCAR–based PCR primers to detect the hybrid pathogen Phytophthora alni and its subspecies causing alder disease in Europe

PCR-based monitoring of recent isolates of tobacco blue mold from Europe reveals the presence of two genetically distinct phenotypes differing in fungicide sensitivity

Development of SCAR Markers for the Identification ofPhytophthora katsuraeCausing Chestnut Ink Disease in Korea

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