Abstract

BackgroundAspergillus westerdijkiae produces ochratoxin A (OTA) in Aspergillus section Circumdati. It is responsible for the contamination of agricultural crops, fruits, and food commodities, as its secondary metabolite OTA poses a potential threat to animals and humans. As a member of the filamentous fungi family, its capacity for enzymatic catalysis and secondary metabolite production is valuable in industrial production and medicine. To understand the genetic factors underlying its pathogenicity, enzymatic degradation, and secondary metabolism, we analysed the whole genome of A. westerdijkiae and compared it with eight other sequenced Aspergillus species.ResultsWe sequenced the complete genome of A. westerdijkiae and assembled approximately 36 Mb of its genomic DNA, in which we identified 10,861 putative protein-coding genes. We constructed a phylogenetic tree of A. westerdijkiae and eight other sequenced Aspergillus species and found that the sister group of A. westerdijkiae was the A. oryzae - A. flavus clade. By searching the associated databases, we identified 716 cytochrome P450 enzymes, 633 carbohydrate-active enzymes, and 377 proteases. By combining comparative analysis with Kyoto Encyclopaedia of Genes and Genomes (KEGG), Conserved Domains Database (CDD), and Pfam annotations, we predicted 228 potential carbohydrate-active enzymes related to plant polysaccharide degradation (PPD). We found a large number of secondary biosynthetic gene clusters, which suggested that A. westerdijkiae had a remarkable capacity to produce secondary metabolites. Furthermore, we obtained two more reliable and integrated gene sequences containing the reported portions of OTA biosynthesis and identified their respective secondary metabolite clusters. We also systematically annotated these two hybrid t1pks-nrps gene clusters involved in OTA biosynthesis. These two clusters were separate in the genome, and one of them encoded a couple of GH3 and AA3 enzyme genes involved in sucrose and glucose metabolism.ConclusionsThe genomic information obtained in this study is valuable for understanding the life cycle and pathogenicity of A. westerdijkiae. We identified numerous enzyme genes that are potentially involved in host invasion and pathogenicity, and we provided a preliminary prediction for each putative secondary metabolite (SM) gene cluster. In particular, for the OTA-related SM gene clusters, we delivered their components with domain and pathway annotations. This study sets the stage for experimental verification of the biosynthetic and regulatory mechanisms of OTA and for the discovery of new secondary metabolites.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2974-x) contains supplementary material, which is available to authorized users.

Highlights

  • Aspergillus westerdijkiae produces ochratoxin A (OTA) in Aspergillus section Circumdati

  • To assess the gene space in the A. westerdijkiae genome, we used a set of 248 core eukaryotic genes to perform CEGMA prediction, which showed that 234 out of 248 (94.35 %) genes were completely matched to our assembled genome

  • We found that the GH3 and AA3 enzymes could be classified into the close phylogenetic profiling clusters summarized in the heatmap of Carbohydrate activity enzyme (CAZyme), which suggested that the AA3 enzymes might act in conjunction with GH3 enzymes

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Summary

Introduction

Aspergillus westerdijkiae produces ochratoxin A (OTA) in Aspergillus section Circumdati. It is responsible for the contamination of agricultural crops, fruits, and food commodities, as its secondary metabolite OTA poses a potential threat to animals and humans. As a member of the filamentous fungi family, its capacity for enzymatic catalysis and secondary metabolite production is valuable in industrial production and medicine. Aspergillus westerdijkiae (CBS 112803 = NRRL 3174), a filamentous fungus branched from the A. ochraceus taxon [1], has a worldwide distribution and mainly colonizes agricultural crops and various food commodities, such as coffee, beer, wine, milk, grapes, oranges, and juice [2,3,4]. Researchers developed a real-time quantitative PCR protocol to detect and quantify A. westerdijkiae contamination in grapes and green coffee beans, focussing on the ITS1-5.8S-ITS2 region within the rDNA unit, which serves as a tag to evaluate A. westerdijkiae contamination and has been frequently used to discriminate at the species level [17]

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