Detection of Banana mild mosaic virus and banana virus X by polyvalent degenerate oligonucleotide reverse transcription PCR (PDO-RT-PCR)

Teycheney Pierre-Yves, Acina Isabelle Nina, Lockhart Benham E.L., Candresse Thierry. 2007. Detection of Banana mild mosaic virus and banana virus X by polyvalent degenerate oligonucleotide reverse transcription PCR (PDO-RT-PCR). In : 11 èmes Rencontres de virologie végétale, Aussois, 28 janvier au 1er février 2007. s.l. : s.n., 1 p. Rencontres de virologie végétale. 11, Aussois, France, 28 Janvier 2007/1 Février 2007.

Résumé : An existing polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) assay [1] was adapted for the detection of Banana mild mosaic virus (BanMMV) and Banana virus X, two Flexiviridae infecting Musa spp. PDO inosine-containing primers were found to be well suited for the detection of BanMMV, despite its high sequence variability [2], but not for that of the highly conserved BVX [3], for which species-specific primers were therefore designed. The sampling and sample processing steps were optimized in order to avoid nucleic acid purification prior to the reverse transcription step. A polyclonal anti-BanMMV antiserum was raised and successfully used for the immunocapture (IC) of BanMMV viral particles from leaf extracts, leading to the development of a PDO-IC-RT-nested PCR assay [4]. Although the anti-BanMMV antiserum could to some extent recognize BVX particles, direct binding (DB) was shown to be a more efficient method for processing BVX-infected samples and a PDO-DB-RT-nested PCR assay was developed for the detection of BVX from leaf extracts [4]. This work completes existing PCR-based detection of other viral species infecting Musa spp. [5, 6] and will benefit movement and propagation of Musa germplasm, for which viruses are important constraints.

Mots-clés Agrovoc : Musa, virus des végétaux, identification, PCR

Classification Agris : H20 - Maladies des plantes
F30 - Génétique et amélioration des plantes

Auteurs et affiliations

• Teycheney Pierre-Yves, CIRAD-BIOS-UPR Multiplication végétative (GLP) ORCID: 0000-0002-9754-0745
• Acina Isabelle Nina, CIRAD-BIOS-UPR Multiplication végétative (GLP)
• Lockhart Benham E.L., University of Minnesota (USA)
• Candresse Thierry, INRA (FRA)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/536864/)

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