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Imaging the invasion of rice roots by the bakanae agent Fusarium fujikuroi using a GFP-tagged isolate

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Abstract

Fusarium fujikuroi (teleomorph Gibberella fujikuroi) is the main seed-borne pathogen of rice, the causal agent of bakanae, a disease that in the last years has become of increasing economical concern in many Italian rice growing areas. A virulent F. fujikuroi isolate was tagged with the green fluorescent protein (gfp) gene using Agrobacterium tumefaciens mediated transformation, and the virulence of the GFP isolate has been confirmed. Little is known about the early interaction of the pathogen with its host, in this work fungal development during the F. fujikuroi/root interaction was analysed by Laser Scanning Confocal Microscopy (LSCM), by using the GFP isolate obtained. The infection of rice roots was investigated from 48 h to 8 days post-inoculation both in resistant and susceptible cultivars. Roots of resistant genotype seem to trigger a hypersensitive response at the infection site and LSCM analysis of root sections allowed the visualization of fungal growth within host tissues. Fungal growth occurred both in the resistant and the susceptible cultivar, even if it was less abundant in the resistant one. Expression analysis of Chitinase1, a gene involved in fungal pathogenesis, was investigated by qPCR on the F. fujikuroi infected rice roots. Chitinase1 expression increased greatly upon infection in the resistant cultivar Selenio.

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Acknowledgements

This work was supported by the AGER Foundation (grant 2010-2369), project RISINNOVA ‘Integrated genetic and genomic approaches for new Italian rice breeding strategies’; by the Ministerio de Ciencia, Innovación y Universidades/FEDER–Agencia Estatal de Investigación (RTI2018-101275-B-I00), the CERCA Programme (“Generalitat de Catalunya”), and MINECO (“Severo Ochoa Programme for Centres of Excellence in R&D” 2016-2019, SEV-2015-0533).

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Authors and Affiliations

  1. Council for Agricultural Research and Economics (CREA), Research Centre for Plant Protection and Certification (CREA-DC), Via C.G. Bertero 22, 00156, Rome, Italy

    Maria Aragona & Alessandro Infantino

  2. Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB, Edifici CRAG, Campus UAB Bellaterra (Cerdanyola del Vallés), 08193, Barcelona, Spain

    Lidia Campos-Soriano & Blanca San Segundo

  3. Department of Agricultural, Forest and Food Sciences (DISAFA) and AGROINNOVA - Centre of Competence, University of Torino, Largo Braccini 2, 10095, Grugliasco, TO, Italy

    Edoardo Piombo & Davide Spadaro

  4. AGROINNOVA – Centre of Competence for the Innovation in the Agro-environmental Sector, University of Turin, Largo Braccini 2, 10095, Grugliasco, TO, Italy

    Edoardo Piombo & Davide Spadaro

  5. Centre of Advanced Microscopy “P. Albertano”, Department of Biology, University of Rome “Tor Vergata”, 00133, Rome, Italy

    Elena Romano

  6. Consejo Superior de Investigaciones Científicas, Barcelona, Spain

    Blanca San Segundo

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  1. Maria Aragona
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  2. Lidia Campos-Soriano
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  3. Edoardo Piombo
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  4. Elena Romano
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  5. Blanca San Segundo
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  6. Davide Spadaro
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  7. Alessandro Infantino
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Contributions

Maria Aragona designed the project, performed the root infection and analysis and wrote the manuscript; Lidia Campos-Soriano performed the genetic transformation of F. fujikuroi; Edoardo Piombo performed the expression analyses and wrote the manuscript; Elena Romano performed LSCM analysis; Alessandro Infantino contributed to design the research work and cared the mycological part; Davide Spadaro and Blanca San Segundo contributed to design the research work and writing the manuscript. All authors commented on previous versions of the manuscript, read and approved the final manuscript.

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Correspondence to Maria Aragona.

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Supplementary Information

Online Resource 1

Fusarium fujikuroi strains used in this study. (XLSX 10 kb)

Online Resource 2

Primers used in this study. (XLSX 10 kb)

Online Resource 3

Phenotype of the GFP-tagged isolate Ff 297-GFP selected for microscopical analyses and the parental strain Ff 297, both grown on PDA plates. (PDF 80 kb)

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Aragona, M., Campos-Soriano, L., Piombo, E. et al. Imaging the invasion of rice roots by the bakanae agent Fusarium fujikuroi using a GFP-tagged isolate. Eur J Plant Pathol 161, 25–36 (2021). https://doi.org/10.1007/s10658-021-02301-z

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