Abstract
An efficient loop-mediated isothermal amplification procedure (LAMP) for the detection of “Candidatus Liberibacter solanacearum” (Lso), the bacterial causal agent of potato zebra chip (ZC) disease, is described in this chapter. Similar to the polymerase chain reaction (PCR), the LAMP employs a bacterial polymerase to amplify specific DNA sequences. However, the method differs from conventional PCR in that it uses six primers specific to the target region to generate a loop structure and autocycling strand displacement rather than thermocycling for sequence amplification. Moreover, unlike PCR that requires agarose gel electrophoresis for resolution, the positive LAMP results can be visualized directly as a precipitate within the reaction tubes. The 16S rDNA gene of “Ca. Liberibacter solanacearum” was used as the target for the design of the six LAMP primers. The LAMP technique is a reliable, rapid, and cost-effective method of detecting the “Ca. Liberibacter solanacearum” pathogen in the potato/tomato psyllid, Bactericera cockerelli, and in field-grown potato plants and tubers.
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Acknowledgments
This work was supported by Texas A&M AgriLife Research Project Number H-8832 (DCG) and Texas Department of Agriculture, Specialty Crop Research, and Product Development Grant Program award SCFB-1213-014. We thank Cecilia Tamborindeguy for providing images of insects and contributions to method development. We also thank J. Creighton Miller Jr. and the Texas Potato Breeding Program for contributions to method development.
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Ravindran, A., Lévy, J., Pierson, E., Gross, D.C. (2015). Loop-Mediated Isothermal Amplification Procedure (LAMP) for Detection of the Potato Zebra Chip Pathogen “Candidatus Liberibacter solanacearum”. In: Lacomme, C. (eds) Plant Pathology. Methods in Molecular Biology, vol 1302. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2620-6_7
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DOI: https://doi.org/10.1007/978-1-4939-2620-6_7
Publisher Name: Humana Press, New York, NY
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