title: A novel and highly efficient method for genetic transformation of fungi employing shock waves.
abstract: Genetic transformation of filamentous fungi is an essential tool in many areas such as biotechnology, medicine, phytopathology and genetics. However, available protocols to transform fungi are inefficient, laborious and have low reproducibility. We report the use of underwater shock waves as a novel method to transform filamentous fungi. An experimental piezoelectric shock wave generator was designed to expose fungal conidia to heterologous DNA. The device was successfully tested in Aspergillus niger, Fusarium oxysporum, Trichoderma reesei and Phanerochaete chrysosporium. The transformation frequency per number of conidia was between two and four orders of magnitude higher in comparison to previously published methods. For example, the frequency of transformation in A. niger was improved up to 5400-fold as compared with Agrobacterium protocols. Transformation was verified by expression of the green fluorescent protein, PCR and Southern blot. Our method offers new possibilities for fast, easy and efficient genetic manipulation of diverse fungal species.
authors.affiliation: Departamento de Ingeniería Genética de Plantas, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del IPN, 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Denis
authors.initials: D
authors.lastname: Magaña-Ortíz
authors.forename: Nancy
authors.initials: N
authors.lastname: Coconi-Linares
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Ortiz-Vazquez
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernández
authors.forename: Achim M
authors.initials: AM
authors.lastname: Loske
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Gómez-Lim
mesh_terms: D005658:Fungi / Q000235:genetics*; D018014:Gene Transfer Techniques / Q000295:instrumentation*; D005827:Genetics, Microbial / Q000295:instrumentation / Q000379:methods*; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods*; D013172:Spores, Fungal / Q000235:genetics; D013314:Stress, Mechanical; D014170:Transformation, Genetic