The CYP51C gene, a reliable marker to resolve interspecific phylogenetic relationships within the Fusarium species complex and a novel target for species-specific PCR

Fernández-Ortuño, Dolores; Loza-Reyes, Elisa; Atkins, Sarah; Fraaije, Bart A.

doi:10.1016/j.ijfoodmicro.2010.10.013

Cited by 34 publications

(28 citation statements)

References 54 publications

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“…The latest findings suggest that sequences which are unique at a species (or group of species) level (e.g. mycotoxin biosynthetic genes) serve as better targets for designing species-specific markers (Alexander et al 2011; Fernández-Ortuño et al 2010; Niessen et al 2004; Proctor et al 2009; Stępień et al 2011a). …”

Section: Discussionmentioning

confidence: 99%

Polymorphism of mycotoxin biosynthetic genes among Fusarium equiseti isolates from Italy and Poland

2012

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Fusarium equiseti (Corda) Saccardo is a soil saprophyte and a weak pathogen, associated with several diseases of fruit and other crops in subtropical and tropical areas, but also in countries with temperate climate. A wide range of secondary metabolites has been identified among natural F. equiseti populations, with zearalenone (ZEA), fusarochromanone and fusarenon-X being the most common. In present study, the genetic diversity of strains from two populations (from Italy and Poland) was evaluated by analysing the translation elongation factor 1α (tef-1α) sequences, two polyketide synthases from the ZEA biosynthetic pathway (PKS13 and PKS4) and the TRI5 gene from the trichothecene biosynthetic pathway. ZEA was produced in rice cultures by 20 of the 27 tested isolates in concentrations ranging from 1.34 ng/g to 34,000 ng/g). The ability to produce enniatins and trichothecenes was evaluated in all strains by identifying esyn1, TRI13 and TRI4 genes. The presence of PKS4 and PKS13 genes was confirmed by polymerase chain reaction (PCR) in only some ZEA-producing isolates. Similarly, the TRI5 gene was found in 14 of the 27 isolates tested. This is likely to have been caused by the divergence of those genes between F. equiseti and F. graminearum (the latter species was used for the primers design) and can be exploited in phylogenetic studies. The analysis of the mycotoxin biosynthetic gene sequences can be used to differentiate the studied genotypes even more precisely than the analysis of the non-coding regions (like tef-1α).

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Section: Discussionmentioning

confidence: 99%

Polymorphism of mycotoxin biosynthetic genes among Fusarium equiseti isolates from Italy and Poland

2012

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“…The CYP51A, CYP51B, and CYP51C genes, which are 1,574, 1,749, and 1,655 nt in length and encode proteins of 507, 526, and 517 aa, respectively (37), are transcribed both in mycelium and fungal conidia. The CYP51C gene is found exclusively in Fusarium species, and is ubiquitous across the genus (38), whereas all other species in the subphylum Pezizomycotina with multiple CYP51 paralogs possess CYP51A and CYP51B genes, with duplications of CYP51A or CYP51B generating the third paralog in some species. Phylogenetic studies have revealed that the amino acid sequence of FgCYP51A is 64.8% identical to MgCYP51A of Magnaporthe grisea, FgCYP51B is 77.7% identical to NcCYP51 from Neurospora crassa, and FgCYP51C shows 80.9% identity to FsCYP51 of Fusarium solani (37).…”

Section: Significancementioning

confidence: 99%

Host-induced gene silencing of cytochrome P450 lanosterol C14α-demethylase–encoding genes confers strong resistance to Fusarium species

Koch¹,

Kumar²,

Weber

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

369

332

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Head blight, which is caused by mycotoxin-producing fungi of the genus Fusarium, is an economically important crop disease. We assessed the potential of host-induced gene silencing targeting the fungal cytochrome P450 lanosterol C-14α-demethylase (CYP51) genes, which are essential for ergosterol biosynthesis, to restrict fungal infection. In axenic cultures of Fusarium graminearum, in vitro feeding of CYP3RNA, a 791-nt double-stranded (ds)RNA complementary to CYP51A, CYP51B, and CYP51C, resulted in growth inhibition [half-maximum growth inhibition (IC 50 ) = 1.2 nM] as well as altered fungal morphology, similar to that observed after treatment with the azole fungicide tebuconazole, for which the CYP51 enzyme is a target. Expression of the same dsRNA in Arabidopsis and barley rendered susceptible plants highly resistant to fungal infection. Microscopic analysis revealed that mycelium formation on CYP3RNA-expressing leaves was restricted to the inoculation sites, and that inoculated barley caryopses were virtually free of fungal hyphae. This inhibition of fungal growth correlated with in planta production of siRNAs corresponding to the targeted CYP51 sequences, as well as highly efficient silencing of the fungal CYP51 genes. The high efficiency of fungal inhibition suggests that host-induced gene-silencing targeting of the CYP51 genes is an alternative to chemical treatments for the control of devastating fungal diseases.T he diseases Fusarium head blight (FHB) and root rot, caused by pathogenic ascomycete fungi of the genus Fusarium, such as Fusarium graminearum (Fg), are devastating diseases of cereal crops (1). Fusarium represents one of the most important cereal killers worldwide, exerting great economic and agronomic impact on global grain production and the grain industry. In addition to considerable yield losses, food quality is detrimentally affected by grain contamination with mycotoxins, which are produced by the fungi during plant infection (2-4). These contaminants represent a serious threat to human and animal health (5).Plant protection and toxin reduction strategies are presently mediated by chemical treatments, resistance breeding strategies, biological control, and genetic engineering. The latter relies on the use of antifungal transgenes, such as chitinase, defensins, polygalacturonase, and the use of mycotoxin detoxifying enzymes (6). However, the use of antifungal traits has not provided convincing practical solutions in terms of efficiency and reliability under agronomical practice.Currently, the application of systemic fungicides, such as sterol demethylation inhibitors (DMIs), is essential for controlling Fusarium diseases and thereby reaching the attainable production level of modern high-yield cultivars. DMI fungicides, such as tebuconazole, triadimefon, and prochloraz, act as ergosterol biosynthesis inhibitors because of cytochrome P450 lanosterol C-14α-demethylase (CYP51) binding, which subsequently disturbs fungal membrane integrity (7). Because of a shortage of alternative chemicals, D...

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“…Uniquely, the third CYP51 paralogue in Fusarium species forms a distinct clade, CYP51C . The CYP51C gene is found exclusively in Fusarium species, and is ubiquitous across the genus, as demonstrated by its use as a reliable phylogenetic marker (Fernández‐Ortuño et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Characterization of the sterol 14α‐demethylases of Fusarium graminearum identifies a novel genus‐specific CYP51 function

et al. 2013

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SummaryCYP51 encodes the cytochrome P450 sterol 14a-demethylase, an enzyme essential for sterol biosynthesis and the target of azole fungicides. In Fusarium species, including pathogens of humans and plants, three CYP51 paralogues have been identified with one unique to the genus. Currently, the functions of these three genes and the rationale for their conservation within the genus Fusarium are unknown.Three Fusarium graminearum CYP51s (FgCYP51s) were heterologously expressed in Saccharomyces cerevisiae. Single and double FgCYP51 deletion mutants were generated and the functions of the FgCYP51s were characterized in vitro and in planta.FgCYP51A and FgCYP51B can complement yeast CYP51 function, whereas FgCYP51C cannot. FgCYP51A deletion increases the sensitivity of F. graminearum to the tested azoles. In DFgCYP51B and DFgCYP51BC mutants, ascospore formation is blocked, and eburicol and two additional 14-methylated sterols accumulate. FgCYP51C deletion reduces virulence on host wheat ears.FgCYP51B encodes the enzyme primarily responsible for sterol 14a-demethylation, and plays an essential role in ascospore formation. FgCYP51A encodes an additional sterol 14a-demethylase, induced on ergosterol depletion and responsible for the intrinsic variation in azole sensitivity. FgCYP51C does not encode a sterol 14a-demethylase, but is required for full virulence on host wheat ears. This is the first example of the functional diversification of a fungal CYP51.

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The CYP51C gene, a reliable marker to resolve interspecific phylogenetic relationships within the Fusarium species complex and a novel target for species-specific PCR

Cited by 34 publications

References 54 publications

Polymorphism of mycotoxin biosynthetic genes among Fusarium equiseti isolates from Italy and Poland

Polymorphism of mycotoxin biosynthetic genes among Fusarium equiseti isolates from Italy and Poland

Host-induced gene silencing of cytochrome P450 lanosterol C14α-demethylase–encoding genes confers strong resistance to Fusarium species

Characterization of the sterol 14α‐demethylases of Fusarium graminearum identifies a novel genus‐specific CYP51 function

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