Comparison of DNA extraction methods for detection of citrus huanglongbing in Colombia

Published

2014-01-01

Comparison of DNA extraction methods for detection of citrus huanglongbing in Colombia

Comparisión de métodos de extracción ADN para detección de huanglongbing en Colombia

Keywords:

Candidatus Liberibacter, Diaphorina citri, Psyllidae, vectors (en)
Candidatus Liberibacter, Diaphorina citri, Psyllidae, vectores (es)

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Authors

Jorge Evelio Angel Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center
Erick Geovanni Hernández Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center
Néstor Andrés Herrera Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center
Linda Yhiset Gómez Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center
Ángela Patricia Castro Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center
Adriana Milena Sepúlveda Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center
Everth Emilio Ebratt Instituto Colombiano Agropecuario (ICA) - National Phytosanitary Laboratory Diagnosis - Tibaitatá Research Center

Abstract (en)
Abstract (es)

Four DNA citrus plant tissue extraction protocols and three methods of DNA extraction from vector psyllid Diaphorina citriKuwayama (Hemiptera: Psyllidae) were compared as part of the validation process and standardization for detection of huanglongbing (HLB). The comparison was done using several criterias such as integrity, purity and concentration. The best quality parameters presented in terms of extraction of DNA from plant midribs tissue of citrus, were cited by Murray and Thompson (1980) and Rodríguez et al. (2010), while for the DNA extraction from psyllid vectors of HLB, the best extraction method was suggested by Manjunath et al.(2008).

En el proceso de validación y estandarización para la detección de huanglongbing (HLB) se compararon protocolos de extracción de ADN, cuatro a partir de tejido vegetal de cítricos y tres a partir del psílido vector Diaphorina citri Kuwayama (Hemiptera: Psyllidae). Para la comparación de los protocolos se utilizaron los criterios de integridad, pureza y concentración. Los protocolos citados por Murray and Thompson (1980) y Rodríguez et al. (2010) fueron los que presentaron los mejores parámetros de calidad en términos de extracción de ADN a partir de la nervadura central del tejido vegetal de cítricos. En la extracción de ADN a partir del psílido vector de HLB, el método que mejores resultados arrojó fue el reportado por Manjunath et al. (2008).

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References

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