Spanu, Francesca (2012) Functional analysis of the Fusarium culmorum genome: gene silencing, transposon tagging and gene knock-out by homologous recombination. Doctoral Thesis.
F. culmorum is an important cereal pathogen having a widespread distribution and able to produce mycotoxins dangerous to humans and animals. The genome of F. culmorum is being sequenced but for many genes the function is yet unknown. A high-throughput strategy for gene identification and for functional genomics analysis is needed. In this project we evaluated the efficiency of 3 complementary approaches to explore the F. culmorum genome. A silencing approach was adopted to downregulating TRI6 gene which regulates the expression of multiple biosynthetic genes. This demonstrated that the type B trichothecenes play an important role as virulence factors in the crown and foot rot (CFR) disease of durum wheat caused by F. culmorum. Hence, this biosynthetic pathway can be regarded as a suitable molecular target for the development of new CFR control tools. The deletion of FcstuA, a member of APSES protein in F. culmorum, enabled to understand its role in CFR pathogenicity, in the biosynthesis of pectolytic enzymes such as polygalacturonase and in the main metabolic or morphological processes regulating the biological cycle of the fungus. Random mutagenesis performed by the mimp1/impala double-component system allowed a complete vision of F. culmorum genome suggesting a valuable exploratory role of this tool. Our results demonstrate that the 3 approaches are useful tools to improve the knowledge on F. culmorum genome and offer new insights for future studies.
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