A multiplex real-time PCR method using hybridization probes for the detection and the quantification of Fusarium proliferatum, F. subglutinans, F. temperatum, and F. verticillioides

Scauflaire, Jonathan; Godet, Marie; Gourgue, Mélanie; Liénard, Charlotte; Munaut, Françoise

doi:10.1016/j.funbio.2012.07.011

Cited by 28 publications

(16 citation statements)

References 22 publications

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“…Some assays are described in the present work (F. temperatum and F. fujikuroi), whereas other protocols were previously described by our group for their use in detecting F. proliferatum [27] and F. verticillioides [28]. The specific primer pair for F. subglutinans was designed by Scauflaire et al (2012) [29], although it was used with the amplification protocol described for F. temperatum (Section 2.2). PCR protocols were also applied to detect the trichothecenes (TCT)-and zearalenone (ZEA)-producing Fusarium species.…”

Section: Detection Aspergillus and Fusarium Species By Specific Pcr mentioning

confidence: 99%

A Comprehensive Study on the Occurrence of Mycotoxins and Their Producing Fungi during the Maize Production Cycle in Spain

et al. 2020

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Mycotoxin contamination is one of the main problems affecting corn production, due to its significant risk to human and animal health. The Fusarium and Aspergillus species are the main producers of mycotoxins in maize, infecting both pre-harvest and during storage. In this work, we evaluated the presence of mycotoxins and their producing species along maize production cycles in three different stages (anthesis, harvest, and storage) during three consecutive seasons (2016–2018). Fungal occurrences were studied using species-specific PCR protocols, whereas mycotoxin levels were determined by LC-MS/MS. Fumonisin-producing Fusarium species (F. verticillioides and F. proliferatum), as well as the aflatoxin producer Aspergillus flavus, were the most predominant species at all stages; although, during some seasons, the presence of F. graminearum and A. niger aggregate species were also identified. Contrastingly, fumonisins were the only mycotoxins detected and levels were always under legal regulations. The results presented here demonstrate that even when fungal contamination occurs at the early stages of the maize production cycle, the application of good agricultural and storage practices might be crucial to ensure mycotoxin-free grains.

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Section: Detection Aspergillus and Fusarium Species By Specific Pcr mentioning

confidence: 99%

A Comprehensive Study on the Occurrence of Mycotoxins and Their Producing Fungi during the Maize Production Cycle in Spain

et al. 2020

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“…in P. sativum roots (Zitnick‐Anderson et al ., ), and several Fusarium spp. in maize (Scauflaire et al ., ; Preiser et al ., ). However, a multiplex qPCR assay designed to quantify the DNA of multiple Fusarium spp.…”

Section: Introductionmentioning

confidence: 99%

Detection of interactions between the pea root rot pathogens Aphanomyces euteiches and Fusarium spp. using a multiplex qPCR assay

et al. 2018

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Pea root rot complex (PRRC) describes a group of closely associated soilborne pathogens that cause root rot disease in field pea. Aphanomyces euteiches and several Fusarium spp. are the most prevalent and damaging microorganisms within this complex, although the impact of interspecific interactions on disease progression remains largely unexplored. Furthermore, a fast and reliable method of detecting and quantifying these pathogens is not currently available. The objectives of this experiment were to: (i) investigate the effect of microbial interactions on root rot severity in pea under greenhouse conditions; and (ii) characterize changes in colonization rates when multiple pathogens are present using qPCR. Seeds were exposed to three species of Fusarium and were planted into A. euteiches‐infested soil in varying combinations. For each experimental treatment, an index of disease severity was used to visually rate disease symptoms. Additionally, two triplex quantitative PCR (qPCR) assays were designed to detect and quantify changes in pathogen population dynamics on the roots. Both assays demonstrated a high degree of sensitivity and efficiency. Results from two independent greenhouse trials indicated an increase in disease severity in the presence of multiple pathogen species compared to single inoculations. Specifically, roots infected with A. euteiches were more susceptible to fusarium root rot than those exposed only to Fusarium spp. These observations were confirmed by qPCR results, which revealed significant changes in colonization rates when multiple species were present. These findings suggest an increased risk of yield loss in regions where A. euteiches and Fusarium spp. co‐occur.

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“…A good correlation between the crossing point value and the DNA content was determined with the r 2 > 0.99 for all the studied Fusarium species. The assay was also successfully applied for the qualitative and quantitative analysis of Fusarium species in contaminated maize samples . Li et al conducted a real‐time PCR assay based on the FOW1 gene encoding a mitochondrial carrier protein for detection and quantification of F. oxysporum , by which a strong positive correlation was found between the copy number (×10) of the F. oxysporum FOW1 gene in 5 g of soil and the colony forming unit (CFU) number .…”

Section: The Use Of Polymerase Chain Reaction For Detecting Fusarium mentioning

confidence: 99%

“…The assay was also successfully applied for the qualitative and quantitative analysis of Fusarium species in contaminated maize samples. 39 Li et al conducted a real-time PCR assay based on the FOW1 gene encoding a mitochondrial carrier protein for detection and quantification of F. oxysporum, 40 by which a strong positive correlation was found between the copy number (×10) of the F. oxysporum FOW1 gene in 5 g of soil and the colony forming unit (CFU) number. 41 Furthermore, a DNA microarray for easy and fast detection of trichothecene-and moniliformin-producing Fusarium species has been developed, which has the capture probes designed by the tef-1 sequence.…”

Section: The Use Of Polymerase Chain Reaction For Detecting Fusarium mentioning

confidence: 99%

Molecular strategies for detection and quantification of mycotoxin‐producing Fusarium species: a review

2014

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Fusarium contamination is considered a major agricultural problem, which could not only significantly reduce yield and quality of agricultural products, but produce mycotoxins that are virulence factors responsible for many diseases of humans and farm animals. One strategy to identify toxigenic Fusarium species is the use of modern molecular methods, which include the analysis of DNA target regions for differentiation of the Fusarium species, particularly the mycotoxin-producing Fusarium species such as F. verticillioides and F. graminearum. Additionally, polymerase chain reaction assays are used to determine the genes involved in the biosynthesis of the toxins in order to facilitate a qualitative and quantitative detection of Fusarium-producing mycotoxins. Also, it is worth mentioning that some factors that modulate the biosynthesis of mycotoxins are not only determined by their biosynthetic gene clusters, but also by environmental conditions. Therefore, all of the aforementioned factors which may affect the molecular diagnosis of mycotoxins will be reviewed and discussed in this paper.

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A multiplex real-time PCR method using hybridization probes for the detection and the quantification of Fusarium proliferatum, F. subglutinans, F. temperatum, and F. verticillioides

Cited by 28 publications

References 22 publications

A Comprehensive Study on the Occurrence of Mycotoxins and Their Producing Fungi during the Maize Production Cycle in Spain

A Comprehensive Study on the Occurrence of Mycotoxins and Their Producing Fungi during the Maize Production Cycle in Spain

Detection of interactions between the pea root rot pathogens Aphanomyces euteiches and Fusarium spp. using a multiplex qPCR assay

Molecular strategies for detection and quantification of mycotoxin‐producing Fusarium species: a review

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