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Citrus tristeza virus: Host RNA Silencing and Virus Counteraction

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Book cover Citrus Tristeza Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2015))

Abstract

To dissect the host RNA silencing response incited by citrus tristeza virus (CTV, genus Closterovirus), a (+) ssRNA of ~19300 nt, and the counter reaction deployed by the virus via its three RNA silencing suppressors (RSS), the small RNAs (sRNAs) of three virus-host combinations were deep sequenced. The subsequent analysis indicated that CTV sRNAs (1) constitute more than half of the total sRNAs in the susceptible Mexican lime and sweet orange, while only 3.5% in the restrictive sour orange; (2) are mostly of 21–22 nt, with those of (+) sense predominating slightly; and (3) derive from all the CTV genome, as evidenced by its entire recomposition from viral sRNA contigs but adopt an asymmetric pattern with a hotspot mapping at the 3′-terminal ~2500 nt. The citrus homologues of Arabidopsis Dicer-like (DCL) 4 and 2 most likely generate the 21 and 22 nt CTV sRNAs, respectively, by dicing the gRNA and the 3′ co-terminal sgRNAs and, particularly, their double-stranded forms accumulating in infected cells. The plant sRNA profile, very similar and dominated by the 24 nt sRNAs in the three mock-inoculated controls, displayed a major reduction of the 24 nt sRNAs in Mexican lime and sweet orange, but not in sour orange. CTV infection also influences the levels of certain microRNAs.

The high accumulation of CTV sRNAs in two of the citrus hosts examined suggests that it is not their synthesis, but their function, the target of the RSS encoded by CTV: p25 (intercellular), p23 (intracellular) and p20 (both). The two latter might block the loading of CTV sRNAs into the RNA silencing complex or interfere with it through alternative mechanisms. Of the three CTV RSS, p23 is the one that has been more thoroughly studied. It is a multifunctional RNA-binding protein with a putative Zn finger domain and basic motifs that (1) has no homologues in other closteroviruses, (2) accumulates in the nucleolus and plasmodesmata, (3) regulates the asymmetric balance of CTV (+) and (−) RNA strands, and (4) induces CTV syndromes and stimulates systemic infection in certain citrus species when expressed as a transgene ectopically or in phloem-associated cells.

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Acknowledgments

This research was supported by a grant (Prometeo/2008/121) from the Generalitat Valenciana, Spain, and by a grant (AGL2009-08052) from the Ministerio de Ciencia e Innovación-Fondo Europeo de Desarrollo Regional. S. Ruiz-Ruiz was additionally supported by a postdoctoral contract from the Generalitat Valenciana (APOSTD/2012/020, Program VALi+d).

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Authors and Affiliations

  1. Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia-Consejo Superior de Investigaciones Científicas, Valencia, Spain

    Susana Ruiz-Ruiz, Leandro Peña & Ricardo Flores

  2. Istituto per la Protezione Sostenibile delle Piante, Consiglio Nazionale delle Ricerche, Bari, Italy

    Beatriz Navarro & Francesco Di Serio

  3. Instituto Valenciano de Investigaciones Agrarias, Moncada, Spain

    Leandro Peña

  4. Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Valencia, Spain

    Luis Navarro & Pedro Moreno

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  1. Susana Ruiz-Ruiz
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  2. Beatriz Navarro
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  4. Luis Navarro
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  5. Pedro Moreno
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  6. Francesco Di Serio
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  7. Ricardo Flores
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Correspondence to Ricardo Flores .

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Editors and Affiliations

  1. Department of Phytosanitary Sciences, University of Catania, Science and Technology Park of Sicily, Catania, Italy

    Antonino F. Catara

  2. The S. Talkowski Laboratory, Department of Plant Pathology and Weed Research, The Volcani Center, Agricultural Research Organization, Bet Dagan, Israel

    Moshe Bar-Joseph

  3. Consiglio per la Ricerca in agricoltura e l’analisi dell’Economia Agraria, Centro di Olivicoltura, Frutticoltura e Agrumicoltura (CREA-OFA), Acireale (Catania), Italy

    Grazia Licciardello

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Ruiz-Ruiz, S. et al. (2019). Citrus tristeza virus: Host RNA Silencing and Virus Counteraction. In: Catara, A., Bar-Joseph, M., Licciardello, G. (eds) Citrus Tristeza Virus. Methods in Molecular Biology, vol 2015. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9558-5_14

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