Multiplex PCR to detect four different tomato-infecting pathogens
| Autor: | Luis Kameyama-Kawabe, Gabriela Alejandra Quintero-Vásquez, María Luisa Bazán-Tejeda, Eva Martínez-Peñafiel, Rosa María Bermúdez-Cruz |
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| Rok vydání: | 2012 |
| Předmět: |
biology
fungi Leveillula taurica food and beverages General Medicine Ribosomal RNA biology.organism_classification Microbiology Virology genomic DNA Intergenic region Ascomycota Fusarium Solanum lycopersicum Begomovirus Actinomycetales Multiplex polymerase chain reaction Electrophoresis Polyacrylamide Gel Multiplex Solanum Multiplex Polymerase Chain Reaction Clavibacter michiganensis DNA Primers Plant Diseases |
| Zdroj: | Folia Microbiologica. 58:269-276 |
| ISSN: | 1874-9356 0015-5632 |
| Popis: | This work was aimed to develop a multiplex PCR assay to detect infectious agents such as Clavibacter michiganensis subsp. michiganensis, Fusarium sp, Leveillula taurica, and begomoviruses in tomato (Solanum lycopersicum) plants. Specific primer sets of each pathogen were designed based on intergenic ribosomal RNA sequences for the first three, whereas for begomoviruses, primers were designed based on conserved regions. The design also considered that the length (200-800 bp) of the PCR products was resolvable by electrophoresis; thus 296, 380, 457, and 731 bp fragments for Clavibacter, Fusarium, Leveillula, and begomoviruses, respectively, were considered. PCR conditions were optimized to amplify all the products in a single tube from genomic DNA and circumvent PCR inhibitors from infected plants. Finally, when the multiplex PCR assay was tested with tomato plants infected with any of the four pathogens, specific PCR products confirmed the presence of the pathogens. Optimized PCR multiplex allowed for the accurate and simultaneous detection of Clavibacter, Fusarium, Leveillula, and begomoviruses in infected plants or seeds from tomato. |
| Databáze: | OpenAIRE |
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