A PCR-Based Assay for Detection of Puccinia striiformis f. sp. tritici in Wheat

Zhao, Jiaqi; Wang, X J; Chen, C Q; Huang, Lili; Zhen-sheng, Kang

doi:10.1094/pdis-91-12-1669

Cited by 36 publications

(17 citation statements)

References 19 publications

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“…PCR techniques have been used to detect a number of plant pathogenic fungi both qualitatively and quantitatively (Kageyama et al 2003;Vincelli and Tisserat 2008). Such techniques commonly utilize ITSrDNA to construct PCR primers because of the speciesspecificity of this region (Kim and Knudsen 2008;Okubara et al 2008;Terashima et al 2002;Zhao et al 2007). We previously characterized ITS-rDNA of Ph.…”

Section: Discussionmentioning

confidence: 99%

PCR-based assays to detect and quantify Phomopsis sclerotioides in plants and soil

Shishido

Sato²,

Yoshida

et al. 2009

J Gen Plant Pathol

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We developed polymerase chain reaction (PCR) assays to detect and quantify Phomopsis sclerotioides, the causal agent of black root rot of cucurbits. We used internal transcribed spacers 1 and 2 of the ribosomal DNA (rDNA) from representative isolates to search for target sequences. Primer pairs were selected after testing against 40 fungal isolates including 13 Ph. sclerotioides isolates, 9 Phomopsis isolates other than Ph. sclerotioides, and 18 soilborne fungi that were either pathogenic or nonpathogenic to cucurbits. Conventional PCR assays with the primer pair of CPs-1 (forward) and CPs-2 (reverse) produced target DNA amplicons from all Ph. sclerotioides isolates but none of the other isolates tested. From soil and root samples collected from fields naturally infested with black root rot of cucumber and melon, the CPs-1/CPs-2 primer pair successfully amplified target DNA fragments in conventional PCR assays. Moreover, we applied the CPs-1/CPs-2 primer pair in a real-time PCR assay with SYBR Green I, and PCRamplified products were successfully quantified by reference to a standard curve generated by adding known amounts of target DNA. Target Ph. sclerotioides DNA fragments were similarly detected in artificially inoculated roots of cucumber, melon, pumpkin, and watermelon, but quantities of Ph. sclerotioides DNA in their hypocotyls of the hosts varied as follows: melon C cucumber C watermelon [ pumpkin. These results suggest that Ph. sclerotioides infection is not species-specific but the rate of infection may differ among host species.

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Section: Discussionmentioning

confidence: 99%

PCR-based assays to detect and quantify Phomopsis sclerotioides in plants and soil

Shishido

Sato²,

Yoshida

et al. 2009

J Gen Plant Pathol

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“…No cross-amplification was observed. Zhao et al [2007] designed the PSR/PSF primer set that gave a single product of 169 bp for only P. striiformis f. sp. tritici isolates with a maximum traceability of 0.1 pg of target DNA.…”

Section: Pcr Assaysmentioning

confidence: 99%

“…There are many detection systems for F. sporotrichioides , P. striiformis f. sp. tritici , and Z. tritici [Beck and Ligon, 1995;Consolo et al, 2009;Demeke et al, 2005;Fraaije et al, 1999Fraaije et al, , 2001Gao et al, 2016;Guo et al, 2006;Konstantinova and Yli-Mattila, 2004;Lihua et al, 2008;Niessen et al, 2004;Wang et al, 2008Wang et al, , 2009Wilson et al, 2004;Zhao et al, 2007]. Particularly for Z. tritici , the number of known diagnostic tests is justified by the significance of STB, one of the most important foliar diseases of wheat caused by this fungus [Keon et al, 2007;Shetty et al, 2007].…”

Section: Future Perspectivesmentioning

confidence: 99%

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A Review of Conventional PCR Assays for the Detection of Selected Phytopathogens of Wheat

Kuzdraliński

Kot²,

Szczerba³

et al. 2017

Microb Physiol

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Infection of phyllosphere (stems, leaves, husks, and grains) by pathogenic fungi reduces the wheat yield and grain quality. Detection of the main wheat pathogenic fungi provides information about species composition and allows effective and targeted plant treatment. Since conventional procedures for the detection of these organisms are unreliable and time consuming, diagnostic DNA-based methods are required. Nucleic acid amplification technologies are independent of the morphological and biochemical characteristics of fungi. Microorganisms do not need to be cultured. Therefore, a number of PCR-based methodologies have been developed for the identification of key pathogenic fungi, such as Fusarium spp., Puccinia spp., Zymoseptoria tritici, Parastagonospora nodorum, Blumeria graminis f. sp. tritici, and Pyrenophora tritici-repentis. This article reviews frequently used DNA regions for fungus identification and discusses already known PCR assays for detection of the aforementioned wheat pathogens. We demonstrate that PCR-based wheat pathogen identification assays require further research. In particular, the number of diagnostic tests for Fusarium graminearum, Puccinia spp., and P. tritici-repentis are insufficient.

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“…In field studies, PCRbased methods have been reported to detect the presence of wheat infection even before the physiological symptoms are visible on the plant tissue [Wang et al, 2009;Zhao et al, 2007]. Several PCR-based assays are available for the identification of Pgt [Liu et al, 2014;Wang et al, 2011], Pst [Fraaije et al, 2001;Gao et al, 2016;Lihua et al, 2008;Wang et al, 2008Wang et al, , 2009Zhao et al, 2007], and Pt [Atkins and Clark, 2004]. However, the available diagnostic tools are still limited, and some may even show nonspecific amplification [Atkins and Clark, 2004].…”

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confidence: 99%

Novel PCR Assays for the Detection of Biological Agents Responsible for Wheat Rust Diseases: Puccinia triticina and Puccinia striiformis f. sp. tritici

Kuzdraliński

Kot²,

Szczerba³

et al. 2017

Microb Physiol

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The species Puccinia triticina (Pt) and Puccinia striiformis f. sp. tritici (Pst) are devastating cereal pathogens that cause leaf and stripe rust diseases. We developed PCR assays for the species-specific detection of Pt and Pst, 2 biological agents that cause wheat rust disease. For each pathogen, we validated 3 primer sets that target the second largest subunits of the RNA polymerase II (rpb2) and β-tubulin 1 (tub1) genes. The specificities of the primers were verified using naturally infected plant materials with visual symptoms of disease. All primer sets amplified a single DNA fragment of the expected length. The primer sets LidPr15/16, LidPr1/2, and LidPs13/14 were able to detect small amounts of pure fungal DNA with sensitivities of 0.1, 1, and 10 pg/μL, respectively. A sufficient detection limit (1 pg/μL to 5 ng/μL) was observed for all assays when the sensitivity test was performed with host plant DNA. The study also evaluated the simultaneous detection of both rust pathogens, and the multiplex PCR assay generated amplicons of 240 and 144 bp in length for Pts (LidPs9/10) and Pt (LidPr1/2), respectively.

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A PCR-Based Assay for Detection of Puccinia striiformis f. sp. tritici in Wheat

Cited by 36 publications

References 19 publications

PCR-based assays to detect and quantify Phomopsis sclerotioides in plants and soil

PCR-based assays to detect and quantify Phomopsis sclerotioides in plants and soil

A Review of Conventional PCR Assays for the Detection of Selected Phytopathogens of Wheat

Novel PCR Assays for the Detection of Biological Agents Responsible for Wheat Rust Diseases:<b><i> Puccinia triticina</i></b> and <b><i>Puccinia striiformis</i></b> f. sp. <b><i>tritici</i></b>

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