Identification and functional analysis of brassicicene C biosynthetic gene cluster in Alternaria brassicicola

Minami, Atsushi; Tajima, Naoto; Higuchi, Yusuke; Toyomasu, Tomonobu; Sassa, Takeshi; Kato, Nobuo; Dairi, Tohru

doi:10.1016/j.bmcl.2008.11.108

Cited by 45 publications

(39 citation statements)

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“…Fusicoccanes are diterpenoids with a variety of effects on plant physiology, including opening of stomata [68]. A blastp search was performed of the M. fijiensis fusicoccadiene synthase using the non-redundant protein sequence database through NCBI, which identified the best homolog as being from Alternaria brassicicola with 83 % sequence similarity (Additional file 10: Tables S7C and Additional file 11: Table S8); this enzyme catalyzes the first step in the synthesis of a fusicoccane called brassicicene C [69]. The top homologs (Additional file 11: Table S8) were used to create a phylogenetic tree, which showed a bootstrap value of 100 for the relationship between the M. fijiensis fusicoccadiene synthase and the A. brassicicola and Bipolaris victoriae fusicoccadiene synthases (Additional file 12: Figure S4).…”

Section: Resultsmentioning

confidence: 99%

Transcriptome sequencing of Mycosphaerella fijiensis during association with Musa acuminata reveals candidate pathogenicity genes

Noar

Daub

2016

BMC Genomics

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BackgroundMycosphaerella fijiensis, causative agent of the black Sigatoka disease of banana, is considered the most economically damaging banana disease. Despite its importance, the genetics of pathogenicity are poorly understood. Previous studies have characterized polyketide pathways with possible roles in pathogenicity. To identify additional candidate pathogenicity genes, we compared the transcriptome of this fungus during the necrotrophic phase of infection with that during saprophytic growth in medium.ResultsTranscriptome analysis was conducted, and the functions of differentially expressed genes were predicted by identifying conserved domains, Gene Ontology (GO) annotation and GO enrichment analysis, Carbohydrate-Active EnZymes (CAZy) annotation, and identification of genes encoding effector-like proteins. The analysis showed that genes commonly involved in secondary metabolism have higher expression in infected leaf tissue, including genes encoding cytochrome P450s, short-chain dehydrogenases, and oxidoreductases in the 2-oxoglutarate and Fe(II)-dependent oxygenase superfamily. Other pathogenicity-related genes with higher expression in infected leaf tissue include genes encoding salicylate hydroxylase-like proteins, hydrophobic surface binding proteins, CFEM domain-containing proteins, and genes encoding secreted cysteine-rich proteins characteristic of effectors. More genes encoding amino acid transporters, oligopeptide transporters, peptidases, proteases, proteinases, sugar transporters, and proteins containing Domain of Unknown Function (DUF) 3328 had higher expression in infected leaf tissue, while more genes encoding inhibitors of peptidases and proteinases had higher expression in medium. Sixteen gene clusters with higher expression in leaf tissue were identified including clusters for the synthesis of a non-ribosomal peptide. A cluster encoding a novel fusicoccane was also identified. Two putative dispensable scaffolds were identified with a large proportion of genes with higher expression in infected leaf tissue, suggesting that they may play a role in pathogenicity. For two other scaffolds, no transcripts were detected in either condition, and PCR assays support the hypothesis that at least one of these scaffolds corresponds to a dispensable chromosome that is not required for survival or pathogenicity.ConclusionsOur study revealed major changes in the transcriptome of Mycosphaerella fijiensis, when associating with its host compared to during saprophytic growth in medium. This analysis identified putative pathogenicity genes and also provides support for the existence of dispensable chromosomes in this fungus.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3031-5) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Transcriptome sequencing of Mycosphaerella fijiensis during association with Musa acuminata reveals candidate pathogenicity genes

Noar

Daub

2016

BMC Genomics

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“…A combination of in vitro assays with recombinant enzymes and gene disruption allowed Dairi's group to then establish the complete fusicoccin A pathway [189]. His group also identified and functionally characterized the brassicicene C biosynthetic pathway from A. brassicola, which includes, in addition to a fusicoccadiene synthase (AbFs), a dioxygenase and oxidoreductase as well as five P450s [188,190,191].…”

Section: Fusicoccanes and Other Diterpenoids Made By Monofunctional Ementioning

confidence: 98%

Biosynthesis of Terpenoid Natural Products in Fungi

Schmidt‐Dannert

2014

Advances in Biochemical Engineering/Biotechnology

108

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Tens of thousands of terpenoid natural products have been isolated from plants and microbial sources. Higher fungi (Ascomycota and Basidiomycota) are known to produce an array of well-known terpenoid natural products, including mycotoxins, antibiotics, antitumor compounds, and phytohormones. Except for a few well-studied fungal biosynthetic pathways, the majority of genes and biosynthetic pathways responsible for the biosynthesis of a small number of these secondary metabolites have only been discovered and characterized in the past 5-10 years. This chapter provides a comprehensive overview of the current knowledge on fungal terpenoid biosynthesis from biochemical, genetic, and genomic viewpoints. Enzymes involved in synthesizing, transferring, and cyclizing the prenyl chains that form the hydrocarbon scaffolds of fungal terpenoid natural products are systematically discussed. Genomic information and functional evidence suggest differences between the terpenome of the two major fungal phyla--the Ascomycota and Basidiomycota--which will be illustrated for each group of terpenoid natural products.

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“…We searched the database for a homologue of the cytochrome P450 orf1 gene that had been cloned from Alternaria brassicicola, a producer of brassicicene (BC) C (an FC-related compound); it was confirmed to catalyze 8b-hydroxylation of fusicocca-2,10(14)-diene. [43,44] As the 8b-hydroxylated compound might be a intermediate common to both BC and FC biosynthesis, an FC producer should have an orthologue of the orf1 gene. From a BLAST search, a probable truncated candidate gene (62 % amino acid identity to the BC enzyme) was identified in a 2.9 kb contig region, as expected.…”

Section: Identification Of the Papt Genementioning

confidence: 99%

An Enzyme Catalyzing O‐Prenylation of the Glucose Moiety of Fusicoccin A, a Diterpene Glucoside Produced by the Fungus Phomopsis amygdali

et al. 2012

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Isoprenoids form the largest family of compounds found in nature. Isoprenoids are often attached to other moieties such as aromatic compounds, indoles/tryptophan, and flavonoids. These reactions are catalyzed by three phylogenetically distinct prenyltransferases: soluble aromatic prenyltransferases identified mainly in actinobacteria, soluble indole prenyltransferases mostly in fungi, and membrane-bound prenyltransferases in various organisms. Fusicoccin A (FC A) is a diterpene glycoside produced by the plant-pathogenic fungus Phomopsis amygdali and has a unique O-prenylated glucose moiety. In this study, we identified for the first time, from a genome database of P. amygdali, a gene (papt) encoding a prenyltransferase that reversibly transfers dimethylallyl diphosphate (DMAPP) to the 6'-hydroxy group of the glucose moiety of FC A to yield an O-prenylated sugar. An in vitro assay with a recombinant enzyme was also developed. Detailed analyses with recombinant PAPT showed that the enzyme is likely to be a monomer and requires no divalent cations. The optimum pH and temperature were 8.0 and 50 °C, respectively. K(m) values were calculated as 0.49±0.037 μM for FC P (a plausible intermediate of FC A biosynthesis) and 8.3±0.63 μM for DMAPP, with a k(cat) of 55.3±3.3×10⁻³ s. The enzyme did not act on representative substrates of the above-mentioned three types of prenyltransferase, but showed a weak transfer activity of geranyl diphosphate to FC P.

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Identification and functional analysis of brassicicene C biosynthetic gene cluster in Alternaria brassicicola

Cited by 45 publications

References 16 publications

Transcriptome sequencing of Mycosphaerella fijiensis during association with Musa acuminata reveals candidate pathogenicity genes

Transcriptome sequencing of Mycosphaerella fijiensis during association with Musa acuminata reveals candidate pathogenicity genes

Biosynthesis of Terpenoid Natural Products in Fungi

An Enzyme Catalyzing O‐Prenylation of the Glucose Moiety of Fusicoccin A, a Diterpene Glucoside Produced by the Fungus Phomopsis amygdali

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