Plant Disease (2020) 104, 3002-3009

Maísa Ciampi-Guillardi, Juliana Ramiro, Maria Heloisa Duarte de Moraes, Marina Coan Goldoni Barbieri and Nelson S. Massola Jr (2020)
Multiplex qPCR assay for direct detection and quantification of Colletotrichum truncatum, Corynespora cassiicola, and Sclerotinia sclerotiorum in soybean seeds
Plant Disease 104 (11), 3002-3009
Abstract: Precise diagnosis of plant diseases is one of the most effective tools to minimize yield losses. Colletotrichum truncatum, Corynespora cassiicola, and Sclerotinia sclerotiorum are common soilborne pathogens that affect soybeans all over the world. We developed a multiplex quantitative real-time polymerase chain reaction (qPCR) assay to simultaneously detect and quantify the three pathogens in soybean seeds and to survey their occurrence in the main soybean production areas in Brazil. Species-specific primers and probes for C. truncatum and C. cassiicola were designed based on GAPDH and TEF1 genes, respectively, to be combined with qPCR detection of S. sclerotiorum previously reported. The multiplex qPCR assay was successful in the simultaneous detection of C. truncatum, C. cassiicola, and S. sclerotiorum, along with a host internal control. The four pathogens were detected and quantified in artificially and naturally infested soybean seeds, even in the lowest incidence level tested of 0.0625% or 1 infected seed out of 1,599 healthy ones. From 81 seed samples tested, C. truncatum was the most frequently detected pathogen and with higher incidence levels (0.25 to 0.125%), followed by S. sclerotiorum and C. cassiicola, both with lower incidence levels (0.125 to 0.0625%). Together, the results evidenced the high sensitivity of the multiplex qPCR assay, indicating its usefulness for a quick and reliable diagnosis of soybean diseases in seeds.
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Link to article at publishers website

Research topic(s) for pests/diseases/weeds:
surveys/sampling/distribution
transmission/dispersal of plant diseases

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Sclerotinia sclerotiorum	Soybean (Glycine max)	Brazil (south)
	Colletotrichum truncatum	Soybean (Glycine max)	Brazil (south)
	Corynespora cassiicola	Soybean (Glycine max)	Brazil (south)