Transposition of the miniature inverted-repeat transposable element mimp1 in the wheat pathogen Fusarium culmorum.
High-throughput methods are needed for functional genomics analysis in Fusarium culmorum, the cause of crown and foot rot on wheat and a type B trichothecene producer. Our aim was to develop and test the efficacy of a double-component system based on the ability of the impala transposase to transactivate the miniature inverted-repeat transposable element mimp1 of Fusarium oxysporum. We report, for the first time, the application of a tagging system based on a heterologous transposon and of splinkerette-polymerase chain reaction to identify mimp1 flanking regions in the filamentous fungus F. culmorum. Similar to previous observations in Fusarium graminearum, mimp1 transposes in F. culmorum by a cut-and-paste mechanism into TA dinucleotides, which are duplicated on insertion. mimp1 was reinserted in open reading frames in 16.4% (i.e. 10 of 61) of the strains analysed, probably spanning throughout the entire genome of F. culmorum. The effectiveness of the mimp1/impala double-component system for gene tagging in F. culmorum was confirmed phenotypically for a putative aurofusarin gene. This system also allowed the identification of two genes putatively involved in oxidative stress-coping capabilities in F. culmorum, as well as a sequence specific to this fungus, thus suggesting the valuable exploratory role of this tool.
Dipartimento di Agraria - Sezione di Patologia Vegetale ed Entomologia and Centro interdisciplinare per lo sviluppo della ricerca biotecnologica e per lo studio della biodiversità della Sardegna e dell'area Mediterranea, Università degli Studi di Sassari, Via E. De Nicola 9, I-07100 Sassari, Italy.
Francesca
F
Spanu
Matias
M
Pasquali
Barbara
B
Scherm
Virgilio
V
Balmas
Angela
A
Marcello
Giuseppe
G
Ortu
Marie
M
Dufresne
Lucien
L
Hoffmann
Marie-Josée
MJ
Daboussi
Quirico
Q
Migheli
D015139:Blotting, Southern; D004251:DNA Transposable Elements / Q000235:genetics*; D005670:Fusarium / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics; D055029:Inverted Repeat Sequences / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000235:genetics; D014908:Triticum / Q000382:microbiology*
D004251:DNA Transposable Elements