Evolution of a fumonisin biosynthetic gene cluster in Fusarium fujikuroi species complex

ABSTRACT Nine biological species, or mating populations (MPs), denoted by letters A to I, and at least 29 anamorphic Fusarium species have been identified within the Gibberella fujikuroi species complex. Members of this species complex are the only species of the genus Fusarium that contain the gibberellin (GA) biosynthetic gene cluster or at least parts of it. However, the ability of fusaria to produce GAs is so far restricted to Fusarium fujikuroi, although at least six other MPs contain all the genes of the GA biosynthetic gene cluster. Members of Fusarium proliferatum, the closest related species, have lost the ability to produce GAs as a result of the accumulation of several mutations in the coding and 5′ noncoding regions of genes P450-4 and P450-1, both encoding cytochrome P450 monooxygenases, resulting in metabolic blocks at the early stages of GA biosynthesis. In this study, we have determined additional enzymatic blocks at the first specific steps in the GA biosynthesis pathway of F. proliferatum: the synthesis of geranylgeranyl diphosphate and the synthesis of ent-kaurene. Complementation of these enzymatic blocks by transferring the corresponding genes from GA-producing F. fujikuroi to F. proliferatum resulted in the restoration of GA production. We discuss the reasons for Fusarium species outside the G. fujikuroi species complex having no GA biosynthetic genes, whereas species distantly related to Fusarium, e.g., Sphaceloma spp. and Phaeosphaeria spp., produce GAs.

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Frequency of three mutations in the fumonisin biosynthetic gene cluster of Fusarium fujikuroi that are predicted to block fumonisin production

World Mycotoxin Journal ◽

10.3920/wmj2020.2572 ◽

2021 ◽

Vol 14 (1) ◽

pp. 49-59

Author(s):

S. Sultana ◽

W.X. Bao ◽

M. Shimizu ◽

K. Kageyama ◽

H. Suga

Keyword(s):

Transcription Factor ◽

Secondary Metabolites ◽

Gene Cluster ◽

Biosynthetic Gene Cluster ◽

Pcr Analysis ◽

Fusarium Fujikuroi ◽

Biosynthetic Gene ◽

Transcription Factor Gene ◽

Factor Gene ◽

Group Strain

Fusarium fujikuroi is the most prominent pathogen found in rice. In addition to gibberellin, F. fujikuroi produces various secondary metabolites, including the polyketide mycotoxins, fumonisins. Fumonisin production is conferred by the fumonisin biosynthetic gene (FUM) cluster consisting of 15-17 genes. F. fujikuroi is phylogenetically subclassified into one group with fumonisin production (F-group) and another group in which fumonisin production is undetectable (G-group). In a previous study, a G-to-T substitution (FUM21_G2551T) in the FUM cluster transcription factor gene, FUM21, was identified as a cause of fumonisin-non-production in a G-group strain. In the current study, further analysis of G-group strains identified two additional mutations that involved FUM-cluster genes essential for fumonisin production: (1) a 22.4-kbp deletion in the FUM10-FUM19 region; and (2) a 1.4-kbp insertion in FUM6. PCR analysis of 44 G-group strains, indicated that 84% had the FUM21_G2551T mutation, 50% had the 22.4-kbp FUM10-FUM19 deletion, and 32% had the 1.4-kbp insertion in FUM6, and some strains had two or all the mutations. None of the mutations were detected in the 51 F-group strains examined. Each of the three mutations alone could account for the lack of fumonisin production in G-group strains. However, one G-group strain did not have any of the mutations. Therefore, another mutation(s) is likely responsible for the lack of fumonisin production in some G-group strains of F. fujikuroi.

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Isolation and Characterization of the Gibberellin Biosynthetic Gene Cluster in Sphaceloma manihoticola

Applied and Environmental Microbiology ◽

10.1128/aem.00694-08 ◽

2008 ◽

Vol 74 (17) ◽

pp. 5325-5339 ◽

Cited By ~ 52

Author(s):

Christiane Bömke ◽

Maria Cecilia Rojas ◽

Fan Gong ◽

Peter Hedden ◽

Bettina Tudzynski

Keyword(s):

Gene Cluster ◽

Gene Clusters ◽

Biosynthetic Gene Cluster ◽

Gene Replacement ◽

Gibberella Fujikuroi ◽

Cytochrome P450 Monooxygenases ◽

Fusarium Fujikuroi ◽

Biosynthetic Gene ◽

Isolation And Characterization ◽

Gene Functions

ABSTRACT Gibberellins (GAs) are tetracyclic diterpenoid phytohormones that were first identified as secondary metabolites of the fungus Fusarium fujikuroi (teleomorph, Gibberella fujikuroi). GAs were also found in the cassava pathogen Sphaceloma manihoticola, but the spectrum of GAs differed from that in F. fujikuroi. In contrast to F. fujikuroi, the GA biosynthetic pathway has not been studied in detail in S. manihoticola, and none of the GA biosynthetic genes have been cloned from the species. Here, we present the identification of the GA biosynthetic gene cluster from S. manihoticola consisting of five genes encoding a bifunctional ent-copalyl/ent-kaurene synthase (CPS/KS), a pathway-specific geranylgeranyl diphosphate synthase (GGS2), and three cytochrome P450 monooxygenases. The functions of all of the genes were analyzed either by a gene replacement approach or by complementing the corresponding F. fujikuroi mutants. The cluster organization and gene functions are similar to those in F. fujikuroi. However, the two border genes in the Fusarium cluster encoding the GA4 desaturase (DES) and the 13-hydroxylase (P450-3) are absent in the S. manihoticola GA gene cluster, consistent with the spectrum of GAs produced by this fungus. The close similarity between the two GA gene clusters, the identical gene functions, and the conserved intron positions suggest a common evolutionary origin despite the distant relatedness of the two fungi.

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Anacyclamide D8P, a prenylated cyanobactin from a Sphaerospermopsis sp. cyanobacterium

10.26434/chemrxiv.5346739.v1 ◽

2017 ◽

Cited By ~ 1

Author(s):

Joana Martins ◽

Niina Leikoski ◽

Matti Wahlsten ◽

Joana Azevedo ◽

Jorge Antunes ◽

...

Keyword(s):

Gene Cluster ◽

Cyclic Peptides ◽

Biosynthetic Gene Cluster ◽

The Novel ◽

Tyrosine Residue ◽

Biosynthetic Gene ◽

Post Translational Modification ◽

Biological Assays ◽

Mass Spectrometric Data ◽

Spectrometric Data

Cyanobactins are a family of linear and cyclic peptides produced through the post-translational modification of short precursor peptides. Anacyclamides are macrocyclic cyanobactins with a highly diverse sequence that are common in the genus Anabaena. A mass spectrometry-based screening of potential cyanobactin producers led to the discovery of a new prenylated member of this family of compounds, anacyclamide D8P (1), from Sphaerospermopsis sp. LEGE 00249. The anacyclamide biosynthetic gene cluster (acy) encoding the novel macrocyclic prenylated cyanobactin, was sequenced. Heterologous expression of the acy gene cluster in Escherichia coli established the connection between genomic and mass spectrometric data. Unambiguous establishment of the type and site of prenylation required the full structural elucidation of 1 using Nuclear Magnetic Resonance (NMR), which demonstrated that a forward prenylation occurred on the tyrosine residue. Compound 1 was tested in pharmacologically or ecologically relevant biological assays and revealed moderate antimicrobial activity towards the fouling bacterium Halomonas aquamarina CECT 5000.

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