Colorimetric LAMP Assay for Detection of Xanthomonas phaseoli pv. manihotis in Cassava Through Genomics : A New Approach to an Old Problem

Bacterial blight caused by Xanthomonas phaseoli pv. manihotis (Xpm) is considered the main bacterial disease that affects cassava, causing significant losses when not properly managed. In the present study, a fast, sensitive, and easy-to-apply method to detect Xpm via colorimetric loop-mediated isot...

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Veröffentlicht in:Plant disease. - 1997. - 108(2024), 10 vom: 11. Okt., Seite 2993-3000
1. Verfasser: Bispo Carvalho, Ian C (VerfasserIn)
Weitere Verfasser: Silva Carvalho, Alice Maria, Wendland, Adriane, Rossato, Maurício
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2024
Zugriff auf das übergeordnete Werk:Plant disease
Schlagworte:Journal Article bacterial blight comparative genomics loop-mediated isothermal amplification DNA Primers
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520 |a Bacterial blight caused by Xanthomonas phaseoli pv. manihotis (Xpm) is considered the main bacterial disease that affects cassava, causing significant losses when not properly managed. In the present study, a fast, sensitive, and easy-to-apply method to detect Xpm via colorimetric loop-mediated isothermal amplification (LAMP) was developed. To ensure the use of a unique-to-the-target pathovar core region for primer design, 74 complete genomic sequences of Xpm together with different bacterial species and pathovars were used for comparative genomics. A total of 42 unique genes were used to design 27 LAMP primer sets, from which nine primers were synthesized, and only one (Xpm_Lp1 primer set) showed sufficient efficiency in preliminary tests. The sensitivity, assessed by a serial dilution of the type strain (IBSBF 278) DNA, yielded high sensitivity, detecting up to 100 fg. The LAMP primers showed high specificity, did not cross-react with other bacterial species or other pathovars tested, and amplified only the Xpm isolates. Tests confirmed the high efficiency of the protocol using infected or inoculated macerated cassava leaves without the need for additional sample treatment. The LAMP test developed in this study was able to detect Xpm in a fast, simple, and sensitive way, and it can be used to monitor the disease under laboratory and field conditions 
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700 1 |a Silva Carvalho, Alice Maria  |e verfasserin  |4 aut 
700 1 |a Wendland, Adriane  |e verfasserin  |4 aut 
700 1 |a Rossato, Maurício  |e verfasserin  |4 aut 
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