Multiplex Polymerase Chain Reaction Assay for the Differential Detection of Trichothecene- and Fumonisin-Producing Species of Fusarium in Cornmeal

Bluhm, Burt H.; Flaherty, Joseph E.; Cousin, M. A.; Woloshuk, Charles P.

doi:10.4315/0362-028x-65.12.1955

Cited by 116 publications

(68 citation statements)

References 25 publications

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“…To assess the ability of FFSC isolates to produce fumonisins, the presence of the FUM1 genes was checked by PCR amplification using primer sets Fum-1/Fum-2 (Baird et al, 2008) and Fum-3/Fum-4 (Bluhm et al, 2002). Primer sets Gfmat1a/Gfmat1b and Gfmat2c/Gfmat2d were used to identify the mating type (MAT-1 or -2) of FFSC strains (Steenkamp et al, 2000).…”

Section: Chemotype and Mating Types Identificationmentioning

confidence: 99%

Crops are a main driver for species diversity and the toxigenic potential ofFusarium isolates in maize ears in China

Zhang

Brankovics

Luo

et al. 2016

WMJ

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In recent years increasing demands and the relatively low-care cultivation of the crop have resulted in an enormous expansion of the acreage of maize in China. However, particularly in China, Fusarium ear rot forms an important constraint to maize production. In this study, we showed that members of both the Fusarium fujikuroi species complex (FFSC) and the Fusarium graminearum species complex are the causal agents of Fusarium ear rot in the main maize producing areas in China. Fumonisin producing Fusarium verticillioides was the most prevalent species, followed by fumonisin producing Fusarium proliferatum and 15-acetyldeoxynivalenol producing F. graminearum. Both Fusarium temperatum and Fusarium boothii were identified for the first time in the colder regions in China, extending their known habitats to colder environments. Mating type analysis of the different heterothallic FFSC species, showed that both types co-occur in each sampling site suggestive of the possibility of sexual recombination. Virulence tests with F. boothii (from maize) and F. graminearum from maize or wheat showed adaptation to the host. In addition, F. graminearum seems to outcompete F. boothii in wheat-maize rotations. Based on our findings and previous studies, we conclude that wheat/maize rotation selects for F. graminearum, while a wheat/rice rotation selects for F. asiaticum. In contrast, F. boothii is selected when maize is cultivated without rotation. A higher occurrence of F. temperatum is observed on maize in colder climatological regions in China, while Fusarium meridionale seems restricted to mountain areas. Each of these species has their characteristic mycotoxin profile and deoxynivalenol and fumonisin are the potential threats to maize production in Northern China.

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Section: Chemotype and Mating Types Identificationmentioning

confidence: 99%

Crops are a main driver for species diversity and the toxigenic potential ofFusarium isolates in maize ears in China

Zhang

Brankovics

Luo

et al. 2016

WMJ

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“…When molecular biology techniques such as PCR are used, they employ as template the obtained isolates genomic DNA and not the genomic DNA obtained directly from the food (BLUHM et al, 2002). Therefore, this methodology was also employed in this study.…”

Section: Resultsmentioning

confidence: 99%

Use of the polymerase chain reaction for detection of Fusarium graminearum in bulgur wheat

et al. 2012

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The detection of mycotoxigenic fungi in foodstuff is important because their presence may indicate the possible associated mycotoxin contamination. Fusarium graminearum is a wheat pathogen and a producer of micotoxins. The polymerase chain reaction (PCR) has been employed for the specific identification of F. graminearum. However, this methodology has not been commonly used for detection of F. graminearum in food. Thus, the objective of the present study was to develop a molecular methodology to detect F. graminearum in commercial samples of bulgur wheat. Two methods were tested. In the first method, a sample of this cereal was contaminated with F. graminearum mycelia. The genomic DNA was extracted from this mixture and used in a F. graminearum specific PCR reaction. The F. graminearum species was detected only in samples that were heavily contaminated. In the second method, samples of bulgur wheat were inoculated on a solid medium, and isolates having F. graminearum culture characteristics were obtained. The DNA extracted from these isolates was tested in F. graminearum specific PCR reactions. An isolate obtained had its trichothecene genotype identified by PCR. The established methodology could be used in surveys of food contamination with F. graminearum.

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“…The yield and purity of the DNA isolated from foodstuffs are crucial determining the success of PCR-based microbial examinations, and a number of improvements to DNA extraction procedures have been reported 5,9,14,25) . However, most of these methods require a series of extraction and purification steps and an appreciable quantity of starting material.…”

Section: Discussionmentioning

confidence: 99%

Microbial Diversity in Milled Rice as Revealed by Riosomal Intergenic Spacer Analysis

et al. 2007

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Microbial diversity in milled rice was examined using culture-independent methods. Environmental DNA samples were extracted from commercial products of milled rice, and the intergenic spacer regions between the small and large subunit ribosomal RNA genes were amplified by ribosomal intergenic spacer analysis (RISA). The results indicated the presence of microbial communities in milled rice and subsequent sequencing of RISA amplicons has identified 17 unique microbial sequences. These include several sequences highly similar to known plant-associated microbes, including Pseudomonas fluorescens, Xanthomonas sacchari, and Pseudozyma antarctica, as well as to several uncultured bacteria. In addition, one sequence shows significant similarity to the sequence of Fusarium graminearum, a potent pathogenic fungus in foodstuffs that produces trichothecene mycotoxins. The potential presence of a pathogenic Fusarium fungus in milled rice was also suggested by a diagnostic PCR procedure that detects the genes involved in the production of trichothecene mycotoxins.

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Multiplex Polymerase Chain Reaction Assay for the Differential Detection of Trichothecene- and Fumonisin-Producing Species of Fusarium in Cornmeal

Cited by 116 publications

References 25 publications

Crops are a main driver for species diversity and the toxigenic potential ofFusarium isolates in maize ears in China

Crops are a main driver for species diversity and the toxigenic potential ofFusarium isolates in maize ears in China

Use of the polymerase chain reaction for detection of Fusarium graminearum in bulgur wheat

Microbial Diversity in Milled Rice as Revealed by Riosomal Intergenic Spacer Analysis

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