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Recombinase Polymerase Amplification Assay for Field Detection of Tomato Bacterial Spot Pathogens
- Source :
- Phytopathology. 109(4)
- Publication Year :
- 2018
-
Abstract
- Bacterial spot of tomato is caused by Xanthomonas gardneri, X. euvesicatoria, X. perforans, and X. vesicatoria. Current diagnostic methods for the pathogens are not in-field assays. Recombinase polymerase amplification (RPA) is ideal for in-field detection assays, because it is an isothermal technique that is rapid and more tolerant to inhibitors compared with polymerase chain reaction. Hence, novel RPA probes and primers were designed to amplify regions of the hrcN gene of X. gardneri, X. euvesicatoria, and X. perforans. The X. gardneri RPA is specific to X. gardneri with a detection limit of 106 CFU/ml and detected X. gardneri in lesions from naturally (n = 6) or artificially (n = 18) infected plants. The X. euvesicatoria RPA detects both X. euvesicatoria and X. perforans with a detection limit of 106 CFU/ml and detected both pathogens in plants artificially infected (n = 36) or naturally infected (n = 85) with either X. euvesicatoria or X. perforans. The X. perforans RPA is specific to X. perforans with a detection limit of 107 CFU/ml. Although the X. perforans RPA assay was unable to detect X. perforans from lesions, the X. euvesicatoria RPA was successfully used in field to detect X. perforans from symptomatic field samples (n = 31). The X. perforans RPA was then used to confirm the pathogen in the laboratory. The X. euvesicatoria and X. gardneri RPA is promising for rapid, real-time in-field detection of bacterial spot and one of the first developed among plant pathogenic bacteria.
- Subjects :
- Xanthomonas
Recombinase Polymerase Amplification
Plant Science
Biology
medicine.disease_cause
law.invention
Microbiology
Field detection
Recombinases
chemistry.chemical_compound
Solanum lycopersicum
law
Bacteriology
medicine
Pathogen
Gene
Polymerase chain reaction
DNA Primers
Plant Diseases
Pathogenic bacteria
DNA Fingerprinting
Bacterial Typing Techniques
chemistry
Agronomy and Crop Science
Nucleic Acid Amplification Techniques
DNA
Subjects
Details
- ISSN :
- 0031949X
- Volume :
- 109
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Phytopathology
- Accession number :
- edsair.doi.dedup.....8678c1abe4385c43ed644350f4b20942