title: Efficient transformation of Mycosphaerella fijiensis by underwater shock waves.
abstract: Black leaf streak disease, also known as black Sigatoka, causes dramatic losses in production of banana and plantains fruits. The disease is caused by the pathogenic fungus Mycosphaerella fijiensis (anamorph Pseudocercospora fijiensis; Mycosphaerellaceae). Genetic transformation of M. fijiensis would allow a better understanding of molecular basis of pathogenicity and design novel approaches to control the infection caused by this pathogen. However, transformation of this fungus has not been easy. We report here a protocol for genetic transformation of M. fijiensis employing underwater shock waves and intact conidia. The recombinant strains recovered showed genetic stability over >10 generations. The frequency of transformation obtained was between 75 and 150 times higher than the efficiency reported in the only article published on transformation of M. fijiensis using spheroplasts. This improvement allowed the use of a thousand times less cells than the amount employed before, avoiding the need for cumbersome successive batch cultures. Our protocol is simple, highly efficient, fast and reproducible and together with the available genomes of M. fijiensis and Musa acuminata, it offers new possibilities to study the diverse mechanisms of pathogenesis of the fungus.
authors.affiliation: Departamento de Ingeniería Genética, CINVESTAV Irapuato, Km 9.6 Libramiento Norte Carretera Irapuato-León, 36821 Irapuato, Guanajuato, Mexico.
authors.forename: Lina
authors.initials: L
authors.lastname: Escobar-Tovar
authors.affiliation: Departamento de Ingeniería Genética, CINVESTAV Irapuato, Km 9.6 Libramiento Norte Carretera Irapuato-León, 36821 Irapuato, Guanajuato, Mexico.
authors.forename: Denis
authors.initials: D
authors.lastname: Magaña-Ortíz
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Blvd. Juriquilla 3001, 76230 Juriquilla, Querétaro, Mexico.
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernández
authors.affiliation: Dirección de Investigaciones, Sección de Fitopatología, Corporación Bananera Nacional, 390-7210 La Rita, Guápiles, Costa Rica.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Guzmán-Quesada
authors.affiliation: Dirección de Investigaciones, Sección de Fitopatología, Corporación Bananera Nacional, 390-7210 La Rita, Guápiles, Costa Rica.
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Sandoval-Fernández
authors.affiliation: Instituto Tecnológico de Mérida, Av. Tecnológico, 97118 Mérida, Yucatán, Mexico.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Ortíz-Vázquez
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Blvd. Juriquilla 3001, 76230 Juriquilla, Querétaro, Mexico. Electronic address: loske@fata.unam.mx.
authors.forename: Achim M
authors.initials: AM
authors.lastname: Loske
authors.affiliation: Departamento de Ingeniería Genética, CINVESTAV Irapuato, Km 9.6 Libramiento Norte Carretera Irapuato-León, 36821 Irapuato, Guanajuato, Mexico. Electronic address: mgomez@ira.cinvestav.mx.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Gómez-Lim
mesh_terms: D001203:Ascomycota / Q000235:genetics*; D005821:Genetic Techniques; D028521:Musa / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology; D013172:Spores, Fungal / Q000235:genetics; D014170:Transformation, Genetic; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water
keywords: Black Sigatoka; Genetic transformation; Musa spp.; Mycosphaerella fijiensis; Shock waves