Molecular detection of Colletotrichum lindemuthianum in bean seed samples

Stélio Jorge Castro Gadaga, Carolina da Silva Siqueira, José da Cruz Machado

Abstract


Colletotrichum lindemuthianum is the causal agent of anthracnose in common bean, and infected seeds are
the most typical propagation form of the disease. Thus, using common bean seeds free of C. lindemuthianum is crucial to
managing this pest, as well as employing fast and accurate detection techniques to ensure high seed quality. In this study,
both conventional and quantitative PCR techniques (cPCR and qPCR) were used for the detection and quantification of C.
lindemuthianum in samples of common bean seeds. For that, seeds were inoculated by exposing them to fungal colonies for
different periods of time, 0 h, 36 h, 72 h, 108 h and 144 h, each period corresponding to an inoculum potential. Then, they were
mixed with healthy seeds, so incidences of 0.25%, 0.50%, 1%, 10%, and 100% of seeds with different inoculum potentials
were obtained, in samples of 400 seeds. Both cPRC and qPCR techniques were effective in detecting the fungus. With the
cPCR method, the highest sensitivity was recorded in those samples with 10% inoculated seeds with inoculum potential P36.
On the other hand, with the qPCR technique, the highest sensitivity in detecting the fungus was observed in samples with
0.25% inoculated seeds with inoculum potential P36.


Keywords


Phaseolus vulgaris, anthracnose, PCR.

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