Real-time PCR for quantification of eleven individual Fusarium species in cereals

Nicolaisen, Mogens; Supronienė, Skaidrė; Nielsen, Linda Kærgaard; Lazzaro, Irene; Spliid, Niels Henrik; Justesen, Annemarie Fejer

doi:10.1016/j.mimet.2008.10.016

Cited by 254 publications

(187 citation statements)

References 14 publications

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“…langsethiae is frequently detected in grains when using species-specific primers; quantitative PCR offers an efficient estimation of the biomass of individual species. Generally, a good correspondence was noted between DNA of F. langsethiae and the T-2/HT-2 toxins detected in grain samples [43][44][45][46][47].…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of Spontaneous Auxotrophic Mutants in <em>Fusarium langsethiae</em>

Gavrilova¹,

Skritnika²,

Gagkaeva³

2017

Preprint

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Analysis of 49 strains of F. langsethiae originating from northern Europe (Russia, Finland, Sweden, UK, Norway, and Latvia) revealed the presence of spontaneous auxotrophic mutants that reflect natural intraspecific diversity. Our investigations detected that 49.0% of F. langsethiae strains were auxotrophic mutants for biotin, and 8.2% of the strains required thiamine as a growth factor. They failed to grow on vitamin-free media. For both prototrophic and auxotrophic strains, no growth defect was observed in rich organic media. Without essential vitamins, a significant reduction in the growth of the auxotrophic strains results in a decrease of the formation of T-2 toxin and diacetoxyscirpenol. In addition, all analysed F. langsethiae strains were distinguished into two subgroups based on PCR product sizes. According to our results, 26 and 23 strains of F. langsethiae belong to subgroups I and II respectively. We determined that the deletion in the IGS region of the rDNA of F. langsethiae belonging to subgroup II is linked with temperature sensitivity and causes a decrease in strain growth at 30 °C. Four thiamine auxotrophic strains were found in subgroup I, while 21 biotin auxotrophic strains were detected in subgroups II. To the best of our knowledge, the spontaneous mutations in F. langsethiae observed in the present work have not been previously reported.

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Section: Discussionmentioning

confidence: 99%

Identification and Characterization of Spontaneous Auxotrophic Mutants in <em>Fusarium langsethiae</em>

Gavrilova¹,

Skritnika²,

Gagkaeva³

2017

Preprint

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“…This was the average genome size for ten of twelve sequenced Dothidiomycete genomes (March 2011; www.osti.gov/bridge/purl.cover.jsp?purl0/ 1012481-8TJMwv/). Nicolaisen et al (2009) achieved a lower detection limit of 2.5 genome equivalents (0.1 pg) for Fusarium spp. in maize using primers to the elongation factor 1 alpha gene.…”

Section: Discussionmentioning

confidence: 99%

“…Waalwijk et al (2008) developed a qPCR assay based on a mycotoxin biosynthesis gene to detect mycotoxin producing Fusarium verticillioides isolates in maize grain from subsistence farmers in South Africa, and correlated fungal DNA content with mycotoxin levels. Nicolaisen et al (2009) established a SYBR Green qPCR assay based on the elongation factor 1 alpha gene to detect and quantify eleven Fusarium sp. in maize and wheat field material.…”

Section: Introductionmentioning

confidence: 99%

Quantitative phenotyping of grey leaf spot disease in maize using real-time PCR

Korsman

Meisel

Kloppers³

et al. 2011

Eur J Plant Pathol

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“…Avocado roots of two rootstocks (Dusa ® and R0. 12) were submerged for an hour in a five liter container containing zoospore suspension at a concentration of 7.2 x 10 4 /ml (mock inoculated plants were submerged in sterile water) after which they were transplanted into 1.5 liter plastic bags filled with perlite (Chemisphere technologies, Gauteng, South Africa). Once transplanted, the zoospore suspension that was used to infect was divided into even portions and added to treated plants (50 ml per plant).…”

Section: Methodsmentioning

confidence: 99%

“…However, pathogen quantification by these methods is not entirely reliable as calculations of pathogen biomass by microscopy are laborious and results can differ greatly between investigators (12). Chemical methods such as fatty acid ergosterol and carbohydrate chitin are used to determine the amount of a specific bio-molecule either present within pathogen cells or released into the environment (5,17).…”

Section: Introductionmentioning

confidence: 99%

Development of a Nested Quantitative Real-Time PCR for DetectingPhytophthora cinnamomiinPersea americanaRootstocks

2013

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Phytophthora cinnamomi causes Phytophthora root rot (PRR) in avocado (Persea americana), an important disease that causes severe economic losses to the avocado industry globally. To date, no PRR-resistant avocado rootstock variety has been discovered, although certain rootstock varieties have been shown to be more tolerant than others. In this study we developed an accurate, low cost assay for in planta quantification of P. cinnamomi to evaluate disease tolerance. A nested real time PCR assay was developed to sensitively detect pathogen DNA in plant tissues. Root samples from a highly tolerant (Dusa ® ) and less tolerant (R0.12) rootstock were collected at 0, 3, 7, 14 and 21 days after inoculation with P. cinnamomi and used for pathogen quantification. Nested primers developed in this study were specific and sensitive and could detect P. cinnamomi in root tissues. The amount of P. cinnamomi quantified in roots was significantly higher in the less tolerant R0.12 plants when compared to the highly tolerant Dusa ® plants at all time points. This study has confirmed the known status of disease tolerance of Dusa ® and R0.12 avocado rootstocks in a quantitative manner and provides a reliable molecular tool to J. Engelbrecht, 2, Plant Disease assist with industry breeding programs for the selection of PRR-resistant avocado rootstock varieties.

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Real-time PCR for quantification of eleven individual Fusarium species in cereals

Cited by 254 publications

References 14 publications

Identification and Characterization of Spontaneous Auxotrophic Mutants in <em>Fusarium langsethiae</em>

Identification and Characterization of Spontaneous Auxotrophic Mutants in <em>Fusarium langsethiae</em>

Quantitative phenotyping of grey leaf spot disease in maize using real-time PCR

Development of a Nested Quantitative Real-Time PCR for DetectingPhytophthora cinnamomiinPersea americanaRootstocks

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