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The goal of this study was to develop a tool specifically designed to identify iterative polyketide synthases (iPKSs) from predicted fungal proteomes. A fungi-based PKS prediction model, specifically for fungal iPKSs, was developed using profile hidden Markov models (pHMMs) based on two essential iPKS domains, the β-ketoacyl synthase (KS) domain and acyltransferase (AT) domain, derived from fungal iPKSs. This fungi-based PKS prediction model was initially tested on the well-annotated proteome of Fusarium graminearum, identifying 15 iPKSs that matched previous predictions and gene disruption studies. These fungi-based pHMMs were subsequently applied to the predicted fungal proteomes of Alternaria brassicicola, Fusarium oxysporum f.sp. lycopersici, Verticillium albo-atrum and Verticillium dahliae. The iPKSs predicted were compared against those predicted by the currently available mixed-kingdom PKS models that include both bacterial and fungal sequences. These mixed-kingdom models have been proven previously by others to be better in predicting true iPKSs from non-iPKSs compared with other available models (e.g. Pfam and TIGRFAM). The fungi-based model was found to perform significantly better on fungal proteomes than the mixed-kingdom PKS model in accuracy, sensitivity, specificity and precision. In addition, the model was capable of predicting the reducing nature of fungal iPKSs by comparison of the bit scores obtained from two separate reducing and nonreducing pHMMs for each domain, which was confirmed by phylogenetic analysis of the KS domain. Biological confirmation of the predictions was obtained by polymerase chain reaction (PCR) amplification of the KS and AT domains of predicted iPKSs from V. dahliae using domain-specific primers and genomic DNA, followed by sequencing of the PCR products. It is expected that the fungi-based PKS model will prove to be a useful tool for the identification and annotation of fungal PKSs from predicted proteomes., (© 2011 The Authors. Molecular Plant Pathology © 2011 BSPP and Blackwell Publishing Ltd.)

Unlabelled: SUMMARY The rapid global re-emergence of Fusarium head blight disease of wheat and barley in the last decade along with contamination of grains with mycotoxins attributable to the disease have spurred basic research on the fungal causal agent. As a result, Fusarium graminearum quickly has become one of the most intensively studied fungal plant pathogens. This review briefly summarizes current knowledge on the pathogenicity, population genetics, evolution and genomics of Fusarium graminearum., Taxonomy: Based on the sexual state Gibberella zeae (Schwein.) Petch: Superkingdom Eukaryota; Kingdom Fungi; Phylum Ascomycota; Subphylum Pezizomycotina; Class Sordariomycetidae; Subclass Hypocreomycetidae; Order Hypocreales; Family Nectriaceae; Genus Gibberella., Host Range: The pathogen is capable of causing head blight or 'scab' on wheat (Triticum), barley (Hordeum), rice (Oryza), oats (Avena) and Gibberella stalk and ear rot disease on maize (Zea). The fungus also may infect other plant species without causing disease symptoms. Other host genera cited for Gibberella zeae or F. graminearum sensu lato (see below) are Agropyron, Agrostis, Bromus, Calamagrostis, Cenchrus, Cortaderia, Cucumis, Echinochloa, Glycine, Hierochloe, Lolium, Lycopersicon, Medicago, Phleum, Poa, Schizachyrium, Secale, Setaria, Sorghum, Spartina and Trifolium. Disease symptoms and signs: For wheat, brown, dark purple to black necrotic lesions form on the exterior surface of the florets and glume (Fig. 1). Although these lesion symptoms sometimes are referred to as scab, they are not formally related to the hyperplasia and hypertrophic epidermal growth associated with other scab diseases such as apple scab. Peduncles immediately below the inflorescence may become discoloured brown/purple. With time, tissue of the inflorescence often becomes blighted, appearing bleached and tan, while the grain within atrophies. Awns often become deformed, twisted and curved downward. In barley, infections are not always readily apparent in the field. Infected spikelets may show a browning or water-soaked appearance. Infected barley kernels show a tan to dark brown discolouration that can be similar to that caused by other kernel blighting organisms. During prolonged wet periods, pink to salmon-orange spore masses of the fungus are often seen on infected spikelets, glumes and kernels in both wheat and barley. For maize ear rot, infection occurs by way of colonizing silk and thus symptoms first appear at the ear apex. White mycelium, turning pink to red with time, colonizes kernels and may progress basipetally, covering the entire ear., Useful Websites: http://www.broad.mit.edu/annotation/fungi/fusarium/mips.gsf.de/genre/proj/fusarium/ http://www.cdl.umn.edu/scab/gz-consort.html http://www.scabusa.org/