The CD1/CD2 marker for specific detection of Colletotrichum lindemuthianum is an iron transporter pseudogene

Gutiérrez, Pablo; Yepes, Mauricio Salazar; Alzate, Juan F.; Berrouet, Katherin Vanegas; Montoya, Mauricio Marín

doi:10.1590/s1982-56762014000400002

Cited by 6 publications

(1 citation statement)

References 28 publications

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“…Based on this outcome, the cPCR technique can be applied for routine laboratory analyses, because it is easily reproducible and takes little time to provide reliable results. Similar findings of other authors endorse this recommendation (Chen et al, 2007;Gutiérrez et al, 2014;Mesquita et al, 1998;Pacheco et al, 2014).…”

Section: Specificity Sensitivity and Limit Of Detection Of The Convsupporting

confidence: 80%

Molecular detection of Colletotrichum lindemuthianum in bean seed samples

2018

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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.Index terms: Phaseolus vulgaris, anthracnose, PCR.Detecção molecular de Colletotrichum lindemuthianum em amostras de sementes de feijão RESUMO -Colletotrichum lindemuthianum é o agente causal da antracnose do feijoeiro, tendo a semente como uma forma de disseminação. O uso de sementes de feijão livres do patógeno é uma das estratégias de manejo, e o uso de técnicas de detecção precisas e rápidas torna-se indispensável para a análise sanitária prévia de sementes. Neste estudo, as técnicas de PCR convencional (cPCR) e PCR em tempo real (qPCR) foram utilizadas para a detecção e quantificação de C. lindemuthianum em amostras de sementes de feijão. Para isso, sementes foram inoculadas por meio de sua exposição às colônias do fungo por tempos variados, 0, 36, 72, 108, e 144 horas, cada período correspondendo a um potencial de inóculo, P0, P36, P72, P108 e P144. As sementes inoculadas foram misturadas com sementes sadias, obtendo-se incidências de 0,25% 0,50%, 1%, 10% e 100% de sementes para cada potencial de inóculo em amostras de 400 sementes. Tanto a cPRC como a qPCR foram eficazes na detecção do fungo. A partir das amostras de sementes com 10% de incidência e com P36, foi possível detectar o fungo por cPCR. Pela técnica de qPCR o patógeno foi detectado em amostras a partir da incidência de 0,25% no menor potencial de inóculo P36.Termos para indexação: Phaseolus vulgaris, antracnose, PCR.1

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Section: Specificity Sensitivity and Limit Of Detection Of The Convsupporting

confidence: 80%