Published

2014-09-01

Citrus huanglongbing: validation of Real-Time PCR (qPCR) for the detection of Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus in Colombia

Citrus huanglongbing: validación de PCR en tiempo real para la detección de Candidatus Liberibacter asiaticus y Candidatus Liberibacter americanus en Colombia

DOI:

https://doi.org/10.15446/agron.colomb.v32n3.44069

Keywords:

Diaphorina citri, HLB, Psyllidae, qPCR, 16S rDNA (en)
Diaphoroina citri, HLB, Psyllidae, qPCR, 16S rDNA (es)

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Authors

  • Jorge Evelio Ángel Instituto Colombiano Agropecuario (ICA)
  • Erick Geovanni Hernández Instituto Colombiano Agropecuario (ICA)
  • Néstor Andrés Herrera Instituto Colombiano Agropecuario (ICA)
  • Linda Yhiset Gómez Instituto Colombiano Agropecuario (ICA)
  • Ángela Patricia Castro Instituto Colombiano Agropecuario (ICA)
  • Adriana Milena Sepúlveda Instituto Colombiano Agropecuario (ICA)
  • Everth Emilio Ebratt Instituto Colombiano Agropecuario (ICA)
Citrus huanglongbing (HLB) is the most destructive citrus disease. Two of the three known HLB-associated Candidatus Liberibacter species were recently found to be present in the Americas. In this study, eggs, nymphs and adults of Diaphorina citri Kuwayama (Hemiptera: Liviidae) and suspect citrus plant materials were collected in 25 municipalities in the departments of Cundinamarca, Santander, Valle del Cauca, Meta and Quindio (Colombia). The detection sensitivity, specificity and assay performance of the 16S rDNA-based real-time PCR (qPCR) were validated for the field survey of the disease in Colombia. The validation confirmed the reliability and robustness of the real-time PCR method for the detection of HLB bacteria in host citrus plant tissues and the vector D. citri. The diagnosis was performed for Candidatus Liberibacter asiaticus (Ca. L. asiaticus) and for Candidatus Liberibacter americanus (Ca. L. americanus) on 168 citrus plant material samples and 239 insect samples. Neither Ca. L. asiaticus nor Ca. L. americanus were detected in the host plants or insects vector, confirming the absence of the disease in the citrus-producing areas of Colombia.
La enfermedad de los cítricos conocida como huanglongbing (HLB) es considerada como la más destructiva para este cultivo. De las tres especies de Candidatus Liberibacter asociadas a HLB, dos han sido recientemente reportadas en América. En el presente trabajo, huevos, ninfas y adultos de Diaphorina citri Kuwayama (Hemiptera: Liviidae) y material de plantas de cítricos sospechosas fueron colectadas en 25 municipios de los departamentos de Cundinamarca, Santander, Valle del Cauca, Meta y Quindio (Colombia). La detección, sensibilidad, especificidad de los ensayos realizados a partir de la región 16s del ADN ribosomal, mediante la prueba de PCR en tiempo real para la detección de la bacteria causante de HLB, fue validada para el monitoreo de la enfermedad en Colombia. La validación confirmó la confiabilidad y robustez del método de PCR en tiempo real para la detección de la bacteria en tejido de plantas de cítricos y en el insecto vector D. citri y se realizó el diagnóstico para Candidatus Liberibacter asiaticus (Ca. L. asiaticus) y para Candidatus Liberibacter americanus (Ca. L. americanus) en 168 muestras de tejido vegetal y en 239 muestras de insectos. Ninguna de las dos variantes de la bacteria fue detectada en plantas e insectos, confirmando la ausencia de la enfermedad en las áreas citrícolas de Colombia.

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Citrus huanglongbing: validation of Real-Time PCR (qPCR) for the detection of Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus in Colombia. (2014). Agronomía Colombiana, 32(3), 377-389. https://doi.org/10.15446/agron.colomb.v32n3.44069

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