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Inoculum concentration and moisture chamber incubation period for inoculation of Xanthomonas citri subsp. citri, the causal agent of citrus canker

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Abstract

Xanthomonas citri subsp. citri is a plant bacterium that causes citrus canker, an important disease in several citrus-growing areas. Due to its relevance, this pathosystem has been frequently investigated, requiring suitable protocols of inoculation for symptom reproduction. This study aimed to evaluate the effect of the concentration of X. citri and the duration of the moisture chamber period on the incidence and severity of citrus canker symptoms under greenhouse conditions. Inoculum concentrations ranging from 100 to 108 cfu/mL were evaluated using both infiltration and spray-inoculation methods. The effect of the moisture chamber on the development of citrus canker lesions was assessed for 24 or 48 h before and/or after inoculation. The minimum concentrations required to infect 100% of the inoculated leaves using infiltration and spray-inoculation were 104 and 107 cfu/mL, respectively. Intermediate severity levels, e.g., 3.2 to 11.4 and 2.5 to 6.7 lesions/cm2, were observed using 103 – 104 cfu/mL for infiltration and 106 – 107 cfu/mL for spray-inoculation. This severity level is suitable when the number of lesions in a given treatment needs to be determined. Conversely, lower or higher inoculum concentrations resulted in severities either extremely low, with no or very few lesions, or excessively high, with coalesced lesions forming a large necrotic area, respectively. A moisture chamber period of 24 h after spray-inoculation was sufficient for reproducing citrus canker lesions under greenhouse conditions. Suitable inoculum concentrations and the use of a moisture chamber after spray-inoculation of X. citri are critical for the successful reproduction of citrus canker symptoms.

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Data availability

The data that support the findings of this study are not openly available due to reasons of sensitivity and are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors acknowledge the Fund for Citrus Protection (Fundecitrus) for the financial support (project #031687) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq—Brazil) for the research productivity fellowship to F. Behlau (grant #307163/2020-2).

Funding

This work was supported by Fundo de Defesa da Citricultura, (grant #031687) and Conselho Nacional de Desenvolvimento Científico e Tecnológico, (grant #307163/2020-2).

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Conceptualization and methodology: Talita Alves dos Santos, Tamiris Garcia da Silva Marin, and Franklin Behlau; Investigation: Talita Alves dos Santos and Tamiris Garcia da Silva Marin; Data curation: Eduardo Silva Gorayeb and Talita Alves dos Santos; Formal analysis: Eduardo Silva Gorayeb; Writing—Original Draft: Eduardo Silva Gorayeb and Talita Alves dos Santos; Resources: Franklin Behlau; Writing – Review and editing: Franklin Behlau.

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Correspondence to Franklin Behlau.

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dos Santos, T.A., Marin, T.G.d., Gorayeb, E.S. et al. Inoculum concentration and moisture chamber incubation period for inoculation of Xanthomonas citri subsp. citri, the causal agent of citrus canker. Eur J Plant Pathol (2024). https://doi.org/10.1007/s10658-024-02841-0

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  • DOI: https://doi.org/10.1007/s10658-024-02841-0

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