Development of Loop-Mediated Isothermal Amplification Assays for the Detection of Seedborne Fungal Pathogens Fusarium fujikuroi and Magnaporthe oryzae in Rice Seed

Bakanae disease (caused by Fusarium fujikuroi) and rice blast (caused by Magnaporthe oryzae) are two of the most important seedborne pathogens of rice. The detection of both pathogens in rice seed is necessary to maintain high quality standards and avoid production losses. Currently, blotter tests a...

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Veröffentlicht in:Plant disease. - 1997. - 102(2018), 8 vom: 01. Aug., Seite 1549-1558
1. Verfasser: Ortega, Sara Franco (VerfasserIn)
Weitere Verfasser: Tomlinson, Jenny, Hodgetts, Jennifer, Spadaro, Davide, Gullino, Maria Lodovica, Boonham, Neil
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2018
Zugriff auf das übergeordnete Werk:Plant disease
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Calmodulin DNA Primers DNA, Fungal Fungal Proteins Peptide Elongation Factor 1
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520 |a Bakanae disease (caused by Fusarium fujikuroi) and rice blast (caused by Magnaporthe oryzae) are two of the most important seedborne pathogens of rice. The detection of both pathogens in rice seed is necessary to maintain high quality standards and avoid production losses. Currently, blotter tests are used followed by morphological identification of the developing pathogens to provide an incidence of infection in seed lots. Two loop-mediated isothermal amplification assays were developed with primers designed to target the elongation factor 1-α sequence of F. fujikuroi and the calmodulin sequence of M. oryzae. The specificity, sensitivity, selectivity, repeatability, and reproducibility for each assay was assessed in line with the international validation standard published by the European and Mediterranean Plant Protection Organization (PM7/98). The results showed a limit of detection of 100 to 999 fg of DNA of F. fujikuroi and 10 to 99 pg of M. oryzae DNA. When combined with a commercial DNA extraction kit, the assays were demonstrated to be effective for use in detection of the pathogens in commercial batches of infected rice seed of different cultivars, giving results equivalent to the blotter method, thus demonstrating the reliability of the method for the surveillance of F. fujikuroi and M. oryzae in seed-testing laboratories 
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650 4 |a Research Support, Non-U.S. Gov't 
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650 7 |a DNA Primers  |2 NLM 
650 7 |a DNA, Fungal  |2 NLM 
650 7 |a Fungal Proteins  |2 NLM 
650 7 |a Peptide Elongation Factor 1  |2 NLM 
700 1 |a Tomlinson, Jenny  |e verfasserin  |4 aut 
700 1 |a Hodgetts, Jennifer  |e verfasserin  |4 aut 
700 1 |a Spadaro, Davide  |e verfasserin  |4 aut 
700 1 |a Gullino, Maria Lodovica  |e verfasserin  |4 aut 
700 1 |a Boonham, Neil  |e verfasserin  |4 aut 
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