title: Multiplex PCR to detect four different tomato-infecting pathogens.
abstract: 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.
authors.affiliation: Genetics and Molecular Biology Department, Centro de Investigaciones y Estudios Avanzados del IPN, Av IPN 2508. Delegación Gustavo A, Madero, Federal District, México.
authors.forename: Gabriela Alejandra
authors.initials: GA
authors.lastname: Quintero-Vásquez
authors.forename: María Luisa
authors.initials: ML
authors.lastname: Bazán-Tejeda
authors.forename: Eva
authors.initials: E
authors.lastname: Martínez-Peñafiel
authors.forename: Luis
authors.initials: L
authors.lastname: Kameyama-Kawabe
authors.forename: Rosa María
authors.initials: RM
authors.lastname: Bermúdez-Cruz
mesh_terms: D000192:Actinomycetales / Q000235:genetics / Q000302:isolation & purification; D001203:Ascomycota / Q000235:genetics / Q000302:isolation & purification; D053839:Begomovirus / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005670:Fusarium / Q000235:genetics / Q000302:isolation & purification; D018551:Solanum lycopersicum / Q000382:microbiology* / Q000821:virology*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology* / Q000821:virology*
chemical_list: D017931:DNA Primers