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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

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Abstract

A qPCR approach was developed to specifically monitor in soils Fusarium graminearum, the main agent responsible for Fusarium Head Blight, and the biocontrol agent Gliocladium catenulatum J1446 (Prestop®). For both fungi, the amplification efficacy of standard curves obtained by mixing pure fungal DNA and soil background DNA was high (qPCR efficacy>96% with R2 > 0.97) with a linear range from 10−3 ng to 10 ng/μL. Our qPCR method allowed quantifying down to 1 μg of F. graminearum and G. catenulatum J1446 mycelium per g of soil. The strong correlation observed between fungal biomass and quantified DNA (R2 = 0.9927 and 0.9356 for F. graminearum and G. catenulatum J1446, respectively) supported the use of the primers to monitor both fungi in soils. Under our experimental conditions, the ability of Prestop® to reduce F. graminearum growth was significantly higher in autoclaved soil compared to living soils, suggesting that there is an antagonistic effect of the soil microbial communities. In contrast, G. catenulatum J1446 growth was mostly not affected by the presence of F. graminearum and was able to persist in both autoclaved and living soils after 15 days of incubation. These results indicate that our qPCR approach may be used to assess the success of soil colonization by a biocontrol agent and its control efficacy by monitoring the dynamics of the BCA and the targeted pathogen in soil.

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Acknowledgements

We gratefully thank Triskalia as project partners and the farmers for kindly giving us access to their field. We also thank the UBO Culture Collection for kindly providing us with fungal strains.

Funding

This work was supported by the Brittany Region [Grant #13008022 Rhisotox] and the French Association for Research and Technology ANRT [Grant #2014/108]. It was certified by the Foodstuff Cluster of the Future (Valorial).

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Authors and Affiliations

  1. EA 3882, Laboratoire Universitaire de Biodiversité et Écologie Microbienne, IBSAM, ESIAB, Technopôle Brest-Iroise, Université de Brest, 29280, Plouzané, France

    Fabienne Legrand, Adeline Picot, José Francisco Cobo-Díaz, Georges Barbier & Gaétan Le Floch

  2. Lallemand SAS, 4 route de Beaupuy, 31180, Castelmaurou, France

    Fabienne Legrand & Olivier Cor

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  1. Fabienne Legrand
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  2. Adeline Picot
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  3. José Francisco Cobo-Díaz
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  4. Olivier Cor
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  5. Georges Barbier
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  6. Gaétan Le Floch
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Correspondence to Adeline Picot.

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Authors confirm that this study is original and has not been published previously, and is not under consideration for publication elsewhere. All authors have approved the manuscript and agree with submission to this journal. This research does not contain any conflict of interest, nor research involving humans or animals.

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Legrand, F., Picot, A., Cobo-Díaz, J.F. et al. Development of qPCR assays to monitor the ability of Gliocladium catenulatum J1446 to reduce the cereal pathogen Fusarium graminearum inoculum in soils. Eur J Plant Pathol 152, 285–295 (2018). https://doi.org/10.1007/s10658-018-1473-0

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