Genetic markers for the analysis of variability and for production of specific diagnostic sequences in fumonisin-producing strains of Fusarium verticillioides

González-Jaén, M. Teresa; Mirete, Salvador; Patiño, Belén; López‐Errasquín, Elena; Vázquez, Carmen M.

doi:10.1007/978-1-4020-2285-2_7

Cited by 18 publications

(20 citation statements)

References 44 publications

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“…In contrast, a PCR‐based method allows simple, accurate, rapid, and precise detection of potential fumonisin‐producing Fusarium species (Bluhm et al. 2004; González‐Jaén et al. 2004; Baird et al.…”

Section: Introductionmentioning

confidence: 99%

Fumonisin B₁production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Dissanayake

Tanaka

Ito

2009

Letters in Applied Microbiology

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Aims: To test the fumonisin B1‐producing ability of Fusarium proliferatum strains isolated from Welsh onion (Allium fistulosum) plants and seeds of commercial cultivars in Japan and to examine the applicability of PCR‐based assays to discriminate between fumonisin B1‐producing and nonproducing isolates. Methods and Results: Fumonisin B1 levels in 20 Fusarium isolates obtained from Welsh onion plants and seeds of seven commercial cultivars were determined by HPLC. Thirteen of the 20 isolates produced fumonisin B1. PCR assay with FUM1 gene‐specific primers amplified a DNA fragment (700 bp) only from fumonisin‐producing isolates. Conclusions: Fusarium proliferatum isolates that can produce fumonisin B1 were often associated with wilted Welsh onion plants and seeds of some commercial cultivars. The PCR assay with FUM1 gene‐specific primers has the potential to discriminate between fumonisin B1‐producing and nonproducing isolates. Significance and Impact of the Study: This study revealed that F. proliferatum producing fumonisin B1 is associated with Welsh onion plants and that commercial cultivar seeds may be contaminated with the fungus. PCR amplification of FUM1 gene can be a useful tool for the rapid identification of fumonisin B1‐producing F. proliferatum isolates.

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Section: Introductionmentioning

confidence: 99%

Fumonisin B₁production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Dissanayake

Tanaka

Ito

2009

Letters in Applied Microbiology

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“…Preliminary studies showed that RAPD was not sensitive enough to distinguish fumonisin forming fungi (Baird, unpublished data). Therefore, specific primers developed from known sequences of genes for fumonisin forming fungi should be investigated [40,41]. González-Jaén et al [40] compared DNA-based strategies for development of primers for identification of F. verticillioides fumonisin producing and non-producing strains.…”

Section: Introductionmentioning

confidence: 99%

Identification of Select Fumonisin Forming Fusarium Species Using PCR Applications of the Polyketide Synthase Gene and its Relationship to Fumonisin Production in vitro

Baird

Abbas

Windham

et al. 2008

IJMS

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A polymerase chain reaction (PCR) based diagnostic assay was used to develop markers for detection of Fusarium verticillioides (=F. moniliforme), a fumonisin producing fungus in maize tissues. Species-specific primers were designed based on sequence data from the polyketide synthase (PKS) gene (FUM1-previously FUM5) responsible for fumonisin production in fungi. Four sets of oligonucleotide primers were tested for their specificity using 24 strains of F. verticillioides, 10 F. proliferatum, and 12 of other Fusarium species. In addition, 13 species of other fungal genera, from four phyla, were tested as negative controls. Among the four sets, primer set B consistently amplified a 419-bp fragment from the DNA 96% of all F. verticillioides strains and 83% of F. proliferatum. All other fungi tested were negative using primer set B. A total of 38% of the F. verticillioides strains grown on a selective liquid medium produced fumonisin and 92% formed the toxin on standard rice medium. When fumonisin formed in culture, PCR assay using primer set B detected every strain of F. verticillioides, but only amplified 80% of F. proliferatum strains that produced the toxin. PCR detection was consistent at 100 pg/µl concentration of genomic DNA from 4 F. verticillioides strains, but varied at 10 pg/µl. Two duplicate greenhouse tests using artificially inoculated maize plants, had greater levels of F. Int. J. Mol. Sci. 2008, 9555 verticillioides detected after re-evaluting using primer set B than from culturing of the tissues. The molecular protocols described in this study requires only 1 day for completion compared to approximately 10 days for cultural work and morphological determination. In conclusion, conventional PCR assay using primer set B provides a sensitive and accurate detection assay that can be used as a primary or secondary confirmation method for identification and occurrence of F. verticillioides within the maize tissues. However, studies using primer set B for fumonisin production determined by strains of F. verticillioides and F. proliferatum will require further verification.

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“…Many researchers use genes from the FUM cluster as a good additional marker for phylogenetic and taxonomic studies of the fumonisin‐producing Fusarium species (González‐Jaén et al . ; Baird et al . ; Stepien et al .…”

Section: Introductionmentioning

confidence: 99%

Molecular diagnostics on the toxigenic potential of F usarium spp. plant pathogens

et al. 2014

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AimsWe propose and test an efficient and rapid protocol for the detection of toxigenic Fusarium isolates producing three main types of Fusarium-associated mycotoxins (fumonisins, trichothecenes and zearelanone).Methods and ResultsThe novel approach utilizes partially multiplexed markers based on genes essential for mycotoxin biosynthesis (fumonisin—fum6, fum8; trichothecenes—tri5, tri6; zearalenone, zea2) in Fusarium spp. The protocol has been verified by screening a collection of 96 isolates representing diverse species of filamentous fungi. Each Fusarium isolate was taxonomically identified through both molecular and morphological techniques. The results demonstrate a reliable detection of toxigenic potential for trichothecenes (sensitivity 100%, specificity 95%), zearalenone (sensitivity 100%, specificity 100%) and fumonisins (sensitivity 94%, specificity 88%). Both presence and identity of toxin biosynthetic genes were further confirmed by direct sequencing of amplification products.ConclusionsThe cross-species-specific PCR markers for key biosynthetic genes provide a sensitive detection of toxigenic fungal isolates, contaminating biological material derived from agricultural fields.Significance and Impact of the StudyThe conducted study shows that a PCR-based assay of biosynthetic genes is a reliable, cost-effective, early warning system against Fusarium contamination. Its future use as a high-throughput detection strategy complementing chemical assays enables effective targeted application of crop protection products.

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Genetic markers for the analysis of variability and for production of specific diagnostic sequences in fumonisin-producing strains of Fusarium verticillioides

Cited by 18 publications

References 44 publications

Fumonisin B₁production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Fumonisin B₁production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Identification of Select Fumonisin Forming Fusarium Species Using PCR Applications of the Polyketide Synthase Gene and its Relationship to Fumonisin Production in vitro

Molecular diagnostics on the toxigenic potential of F usarium spp. plant pathogens

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Genetic markers for the analysis of variability and for production of specific diagnostic sequences in fumonisin-producing strains of Fusarium verticillioides

Cited by 18 publications

References 44 publications

Fumonisin B1production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Fumonisin B1production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Identification of Select Fumonisin Forming Fusarium Species Using PCR Applications of the Polyketide Synthase Gene and its Relationship to Fumonisin Production in vitro

Molecular diagnostics on the toxigenic potential of F usarium spp. plant pathogens

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Fumonisin B₁production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan

Fumonisin B₁production byFusarium proliferatumstrains isolated fromAllium fistulosumplants and seeds in Japan