Reverse Genetics for Functional Genomics of Phytopathogenic Fungi and Oomycetes

Abstract

Sequencing of over 40 fungal and oomycete genomes has been completed. The next major challenge in modern fungal/oomycete biology is now to translate this plethora of genome sequence information into biological functions. Reverse genetics has emerged as a seminal tool for functional genomics investigations. Techniques utilized for reverse genetics like targeted gene disruption/replacement, gene silencing, insertional mutagenesis, and targeting induced local lesions in genomes will contribute greatly to the understanding of gene function of fungal and oomycete pathogens. This paper provides an overview on high-throughput reverse genetics approaches to decode fungal/oomycete genomes.

Olive – Colletotrichum acutatum: An Example of Fruit-Fungal Interaction

“…Similarly to plants, the knowledge of the pathogens genome is very important to monitor global changes that occur during plant-fungal interactions. Several fungal genomes have been now sequenced, being some of them extremely important in terms of yield lost in key crops such as Botrytis cinerea (grape grey mould, and other host species), Fusarium graminearum (cereal head blight), Fusarium verticillioides (corn seed rot), Magnaporthe oryzae (rice blast), Mycosphaerella fijiensis (banana black leaf streak), Septoria tritici (wheat leaf blotch), Puccinia graminis (cereal rust), Phytophthora ramorum (sudden oak death) and Phytophthora sojae (soybean stem/root rot) (Bhadauria et al, 2009). …”

Post-transcriptional silencing of the SGE1 gene induced by a dsRNA hairpin in Fusarium oxysporum f. sp cubense, the causal agent of Panama disease

“…The construction of DNA vectors containing cassettes that encode for long dsRNA hairpins targeting specific genes, and the subsequent insertion of these cassettes into the genomes of organisms of interest allow for the permanent silencing of the target mRNA. This differs from the transfection of synthetic dsRNAs or small interfering RNAs (siRNAs), which produce transient gene silencing (Krajaejun et al, 2007;Shafran et al, 2008;Bhadauria et al, 2009;Caribé dos Santos et al, 2009;Mumbanza et al, 2013).…”

A Review beyond the borders: Proteomics of microclonial black fungi and black yeasts

“…by microarrays), while a proteomics approach is more direct, transcriptomics investigates the all over change of gene expression at mRNA level. Proteomics leads to more reliable conclusions, since proteins are directly related to biological functions and phenotypes [13].…”