Development of qPCR assays to monitor the ability of Gliocladium catenulatum J1446 to reduce the cereal pathogen Fusarium graminearum inoculum in soils

Legrand, Fabienne; Picot, Adeline; Cobo‐Díaz, José F.; Cor, O.; Barbier, Georges; Floch, Gaétan Le

doi:10.1007/s10658-018-1473-0

Cited by 7 publications

(4 citation statements)

References 43 publications

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“…The authors used universally primed-PCR (UP-PCR) and randomly amplified polymorphic DNA (RAPD) techniques to identify sequence-characterized amplified region (SCAR) markers. Recently, Legrand et al (2018) applied these SCAR markers for J1446 to develop a qPCR assay and monitored the establishment of the BCA in non-sterile and sterilized soils artificially inoculated with F. graminearum . The interaction of the antagonist and the pathogen (also monitored by qPCR, using the RAPD markers of Nicholson et al, 1998) in the two environments, revealed significant growth inhibition of the pathogen by up to 50% when the co-introduced BCA was growing in sterilized soil, demonstrating the usefulness of sensitive quantification methods applicable to different matrices.…”

Section: Discussionmentioning

confidence: 99%

“…Relatively little is known about the survival and distribution of C. rosea after the application and available quantification methods are based on laboratory cultivation techniques (Pan et al, 2013) or on strain-specific molecular markers currently used for quantitative real-time polymerase chain reaction (qPCR) detection in soils (Legrand et al, 2018). Hence, for the monitoring of crop residues and grains, a widely applicable DNA based quantification method on the species level is needed.…”

Section: Introductionmentioning

confidence: 99%

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TaqMan qPCR for Quantification of Clonostachys rosea Used as a Biological Control Agent Against Fusarium graminearum

et al. 2019

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Clonostachys rosea is a biological control agent against Fusarium graminearum in small grain cereals and maize. Infections with F. graminearum do not only reduce the yield but, due to the production of mycotoxins, also affect the entire value chain of food and feed. In addition, production of other secondary metabolites such as hydrophobins, also known as gushing inducers, may cause quality challenges for the malting and brewing industry. Sustainable disease control strategies using C. rosea are treatment of infected residues of the previous crop, direct treatment of the actual cereal crop or post-harvest treatment during malting processes. Follow-up of growth and survival of biocontrol organisms during these different stages is of crucial importance. In the current study, we developed a quantitative real-time PCR detection method that amends the currently available culture-dependent techniques by using TaqMan chemistry with a highly specific primer and probe set, targeting the actin gene. We established a sensitive assay that detects the biological control agent down to 100 genome copies per reaction, with PCR efficiencies between 90 and 100%. The specificity of the assay was confirmed against a panel of 30 fungal and 3 bacterial species including 12 members of the Fusarium head blight complex and DNA of barley, maize and wheat. The DNA of C. rosea was detected in Fusarium -infected maize crop residues that were either treated in the laboratory or in the field with C. rosea and followed its DNA throughout the barley malting process to estimate its growth during grain germination. We used a standardized DNA extraction protocol and showed that C. rosea can be quantified in different sample matrices. This method will enable the monitoring of C. rosea during experiments studying the biological control of F. graminearum on cereal crop residues and on cereal grains and will thus contribute to the development of a new disease control strategy.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

TaqMan qPCR for Quantification of Clonostachys rosea Used as a Biological Control Agent Against Fusarium graminearum

et al. 2019

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“…Fusarium Head Blight (FHB) is currently one of the most important diseases, caused primarily by Fusarium graminearum sensu stricto [1], affecting wheat, barley, and other small grain cereal crops [2,3]. The disease causes yield and quality losses and seriously threatens animal and human health due to grains contaminated with mycotoxins [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

Weeds in Cereal Crop Rotations May Host Fusarium Species That Cause Fusarium Head Blight and Grain Weight Losses in Wheat

et al. 2022

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Fusarium Head Blight (FHB) is one of the most common worldwide wheat and other small grain diseases. The infection is caused by Fusarium graminearum and other related species, which significantly reduce grain yield and contaminate grains with mycotoxins which are harmful for humans and animals. Fusarium pathogen survives the winter well in plant debris left on the field. Weeds around and within crops are alternative hosts of Fusarium fungi when an economically important host plant is not present. This article focuses on the determination of DNA content of Fusarium species (F. graminearum and F. avenaceum) in artificially inoculated wheat plants with isolates from weeds, as well as its influence on the severity of FHB and spring wheat 1000-grain weight under field conditions. Fungal DNA content in grains was evaluated by quantitative real-time PCR. The results showed that the DNA concentration of F. graminearum was significantly higher in the grain than F. avenaceum. The severity of FHB when wheat heads were inoculated with F. graminearum was significantly higher than with F. avenaceum. All F. graminearum strains statistically significantly reduced the weight of spring wheat grains, while F. avenaceum did not affect the weight of 1000 grain. This investigation has shown that weeds in crop rotations are a potential source of FHB infection. However, the severity of the disease is more affected by the Fusarium species than the host plant. This experiment is, to our knowledge, the first report on the estimation of Fusarium DNA content in artificially inoculated wheat plants with isolates from weeds, as well as its comparison with pathogenicity to wheat and its effect on 1000-grain weight.

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“…It has been used for the detection and quantification of various soilborne fungi such as Gliocladium spp. (Legrand et al., 2018), Rhizoctonia spp. (Woodhall, Adams, Peters, Harper, & Boonham, 2013), and Fusarium spp.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of substrates and additives to Trichoderma harzianum development by qPCR

et al. 2020

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Biocontrol strains can activate defense genes and promote plant growth. The aim of this study is to evaluate the development of Trichoderma in substrates with additives by quantitative polymerase chain reaction (qPCR). The T. harzianum, T019 isolated from the Protected Geographical Indication Alubia La Bañeza‐León was tested in peat and vermiculite with and without additives. The development was evaluated by qPCR with the α‐actin gene and a standard curve. Statistical analysis was performed using an analysis of variance and mean completely randomized. The qPCR technique has served to quantify precisely the growth of Trichoderma in the different substrates. T. harzianum develops better in the peat substrates supplemented with cornmeal and with a mixture of cornmeal and bentonite than in any of the vermiculate‐based substrates. T. harzianum development was improved by growing in peat substrates supplemented with cornmeal.

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Development of qPCR assays to monitor the ability of Gliocladium catenulatum J1446 to reduce the cereal pathogen Fusarium graminearum inoculum in soils

Cited by 7 publications

References 43 publications

TaqMan qPCR for Quantification of Clonostachys rosea Used as a Biological Control Agent Against Fusarium graminearum

TaqMan qPCR for Quantification of Clonostachys rosea Used as a Biological Control Agent Against Fusarium graminearum

Weeds in Cereal Crop Rotations May Host Fusarium Species That Cause Fusarium Head Blight and Grain Weight Losses in Wheat

Evaluation of substrates and additives to Trichoderma harzianum development by qPCR

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