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This work was supported by project PID2021-123600OR-C44, funded by MICIU/AEI/https://doi.org/10.13039/501100011033 and by ERDF, EU.

Analysis of institutional authors

Cubero, JaimeAuthor

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November 26, 2024
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A reliable qPCR technique for detecting viable Xanthomonas arboricola pv. pruni cells

Publicated to:Applied Microbiology And Biotechnology. 108 (1): 472- - 2024-12-01 108(1), DOI: 10.1007/s00253-024-13288-y

Authors: Sabuquillo, Pilar; Berruete, Isabel M; Cubero, Jaime; Palacio-Bielsa, Ana

Affiliations

CSIC, Inst Nacl Invest & Tecnol Agr & Alimentaria, Madrid, Spain - Author
Univ Zaragoza, Ctr Invest & Tecnol Agroalimentaria Aragon CITA, Inst Agroalimentario Aragon IA2, Zaragoza, Spain - Author

Abstract

Xanthomonas arboricola pv. pruni (Xap) is the causal agent of bacterial spot of stone fruits and almond (Prunus spp). Detection of Xap is typically carried out using quantitative real-time PCR (qPCR) combined with culture-based isolation. However, qPCR does not differentiate between viable and dead cells, potentially leading to an overestimation of the infective population in a sample. Such overestimation could result in unnecessary phytosanitary measures. The present study aims to develop a specific protocol ideally targeting to detection of only live Xap bacterial cells. To address this challenge, the viable quantitative PCR (v-qPCR) method was evaluated using three nucleic acid-binding dyes: propidium monoazide (PMA), a combination of PMA and ethidium monoazide (EMA), and PMAxx (TM), an improved version of PMA. PMAxx (TM) proved to be the most suitable dye for the detection and quantification of living bacterial cells. This methodology was also evaluated in infected plant material over time and can be considered a rapid and reliable alternative to PCR methods for detecting only those putative infective Xap that may pose a risk for Prunus crops.

Keywords

8-azidoethidiumÁrboles frutalesAzidesBiofilmsColorantesCombinationCut-off ctEnfermedades bacterianasFrutas de huesoIntercalating dyIntercalating dyeLodMicrobial viabilityMonoazidePlant diseasesPropidiumPropidium monoazideProtocoProtocolosPrunusQuantitative pcrReacción en cadena de polimerasa cuantitativaReal-time polymerase chain reactionV-qpcrViable cell detectionXanthomonasXanthomonas arboricola pv. pruni

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Applied Microbiology And Biotechnology due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2024 there are still no calculated indicators, but in 2023, it was in position 42/177, thus managing to position itself as a Q1 (Primer Cuartil), in the category Biotechnology & Applied Microbiology.

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-08-01:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 3.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 2 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 0.5.
  • The number of mentions on the social network X (formerly Twitter): 1 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.
  • Assignment of a Handle/URN as an identifier within the deposit in the Institutional Repository: http://hdl.handle.net/10532/7244

Leadership analysis of institutional authors

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: Last Author (Palacio-Bielsa, Ana).

the author responsible for correspondence tasks has been Palacio-Bielsa, Ana.