Abstract
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.
Highlights
Spread in Brazil, the common bean crop is susceptible to several diseases, such as anthracnose (Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi and Cav.), fusarium wilt (Fusarium oxysporum f. sp. phaseoli), rust (Uromyces phaseoli), and bean blight (Xanthomonas axonopodis pv. phaseoli)
Other isolates obtained from bean seeds, namely Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. phaseoli and Rhizoctonia solani, from the mycological collection of the Laboratory of Seed Pathology at Universidade Federal de Lavras (UFLA), were multiplied in similar conditions to those previously described, but using PDA as culture medium instead
This work showed that the pair of primers used for the conventional PCR (cPCR) process was specific for detecting the isolate of C. lindemuthianum, race 65, but not for other pathogens and saprophytes present on bean seeds
Summary
Spread in Brazil, the common bean crop is susceptible to several diseases, such as anthracnose (Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi and Cav.), fusarium wilt (Fusarium oxysporum f. sp. phaseoli), rust (Uromyces phaseoli), and bean blight (Xanthomonas axonopodis pv. phaseoli). Spread in Brazil, the common bean crop is susceptible to several diseases, such as anthracnose Among those caused by fungi, anthracnose is a great concern nowadays, because of its economic importance, and due to the capacity of devastating crops within a short time, under favorable environmental conditions. The seed is one of the most important forms of survival and long-distance dissemination used by the fungus. This fact increases the importance of the quality analyses of seeds, once it can indicate the presence of C. lindemuthianum in samples. Damasceno Silva et al (2007) report that approximately 50 physiological races of C. lindemuthianum have been identified in Brazil, among which 65, 73, and 81 stand out as the most frequent ones, considering the virulence
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