Early Detection of Asian Soybean Rust Using PCR

Lamour, Kurt; Finley, Ledare; Snover-Clift, K.; Stack, James P.; Pierzynski, Joy; Hammerschmidt, R.; Jacobs, James J.; Byrne, J. M.; Harmon, Philip F.; Vitoreli, Anne; Wisler, G. C.; Harmon, Carrie L.; Lévy, L.; Zeller, Kurt A.; Stone, Christine L.; Luster, Douglas G.; Frederick, Reid D.

doi:10.1094/php-2006-0524-01-rs

Cited by 11 publications

(9 citation statements)

References 12 publications

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“…We documented that SBR symptoms first appeared during mid-reproductive stages (R5 and later) of plant growth when seeds began to develop in pods. This contrasts with previous laboratory and greenhouse studies in which symptoms typically developed 7 to 14 days after inoculation (1,10). SBR has been reported on cotyledons and unifoliolate leaves of young soybean plants under intense disease pressure in Brazil (1).…”

Section: Discussioncontrasting

confidence: 81%

Documentation of an Extended Latent Infection Period by Phakopsora pachyrhizi, the Soybean Rust Pathogen

Ward

Schneider

Robertson

2012

Plant Health Progress

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Phakopsora pachyrhizi, the soybean rust pathogen, overwinters on kudzu in the southern United States. However, even with severely affected kudzu adjacent to soybean fields, disease symptoms do not occur on soybeans until plants are in mid-reproductive stages of growth during mid to late summer. These observations suggested that soybeans are exposed to airborne inoculum of the pathogen long before symptoms occur, and we hypothesized that these plants may be latently infected. This hypothesis was confirmed by using quantitative real-time PCR (qPCR) to detect the rust pathogen in soybean leaves 18 to 60 days before symptoms were observed. Additionally, DNA of P. pachyrhizi increased only slightly until one week before symptoms developed. Results from this study documented that soybeans can become infected by the rust pathogen during early stages of plant growth, but symptoms often develop during the mid-reproductive stages. This extended latent infection period may be an optimum time for fungicide applications. Accepted for publication 12 December 2011. Published 21 March 2012.

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Section: Discussioncontrasting

confidence: 81%

Documentation of an Extended Latent Infection Period by Phakopsora pachyrhizi, the Soybean Rust Pathogen

Ward

Schneider

Robertson

2012

Plant Health Progress

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“…Sample results were negative for ELISA, but results were positive for real-time PCR. Although these results are false positives for this study, they are not surprising since real-time PCR has been shown to detect latent infection and is routinely used to detect random spore deposition from air and rain samplers (6,11).…”

Section: Comparison Of Diagnostic Protocolsmentioning

confidence: 82%

“…PCR and real-time PCR are useful tools for confirmation of the soybean rust pathogens at the species level, especially in Florida and the Gulf Coast where P. meibomiae is most likely to occur. PCR is also useful for early detection before the development of soybean rust symptoms currently used in ASR diagnosis (6). The ELISA kit that was evaluated is not species-specific and requires too much time to be used for field application.…”

Section: Discussionmentioning

confidence: 99%

A Comparative Analysis of Diagnostic Protocols for Detection of the Asian Soybean Rust Pathogen, Phakopsora pachyrhizi

Jurick

Harmon

Marois

et al. 2007

Plant Health Progress

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Plant diagnosticians routinely process soybean rust samples with one or more of the following diagnostic protocols: visual assessment using a dissecting microscope, an enzyme-linked immunosorbant assay (ELISA), and pathogen-specific PCR (conventional and real-time). Results of these three diagnostic protocols and time requirements for diagnosis were collected for samples with signs (sporulation) of soybean rust (presumed positive) and samples without sporulating rust pustules (presumed negative). Data were collected for samples processed in triplicate at six university plant disease clinics. The effects of sample treatment and storage on results of two diagnostic protocols also were examined. Visual diagnosis required the least amount of time (4 min), ELISA and PCR took approximately six-and-a-half times longer per sample (multiple samples being processed at one time) than a visual diagnosis. ELISA could not detect the pathogen in positive samples after desiccation followed by long-term storage at room temperature and following one or more autoclave treatments for 30 min. However, conventional PCR was capable of detecting P. pachyrhizi-infected plant material following all sample storage regimes. Accepted for publication 9 February 2007. Published 31 May 2007.

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“…In this case, scouting teams are deployed to survey cropland and surrounding areas. Symptomatic plants are evaluated in diagnostic labs where microscopes are used to assess potential infections and PCR is used to amplify soybean rust-specific DNA [2]. Reports of positive findings are sent back to scientists and growers who work together to develop disease progression models and prepare a response plan that may include spraying fungicides.…”

Section: Introductionmentioning

confidence: 99%

Tandem mass spectrometry for the detection of plant pathogenic fungi and the effects of database composition on protein inferences

et al. 2007

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LC-MS/MS has demonstrated potential for detecting plant pathogens. Unlike PCR or ELISA, LC-MS/MS does not require pathogen-specific reagents for the detection of pathogen-specific proteins and peptides. However, the MS/MS approach we and others have explored does require a protein sequence reference database and database-search software to interpret tandem mass spectra. To evaluate the limitations of database composition on pathogen identification, we analyzed proteins from cultured Ustilago maydis, Phytophthora sojae, Fusarium graminearum, and Rhizoctonia solani by LC-MS/MS. When the search database did not contain sequences for a target pathogen, or contained sequences to related pathogens, target pathogen spectra were reliably matched to protein sequences from nontarget organisms, giving an illusion that proteins from nontarget organisms were identified. Our analysis demonstrates that when database-search software is used as part of the identification process, a paradox exists whereby additional sequences needed to detect a wide variety of possible organisms may lead to more cross-species protein matches and misidentification of pathogens.

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Early Detection of Asian Soybean Rust Using PCR

Cited by 11 publications

References 12 publications

Documentation of an Extended Latent Infection Period by Phakopsora pachyrhizi, the Soybean Rust Pathogen

Documentation of an Extended Latent Infection Period by Phakopsora pachyrhizi, the Soybean Rust Pathogen

A Comparative Analysis of Diagnostic Protocols for Detection of the Asian Soybean Rust Pathogen, Phakopsora pachyrhizi

Tandem mass spectrometry for the detection of plant pathogenic fungi and the effects of database composition on protein inferences

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