Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection

Spiroplasma citri is a phloem-limited bacterium that causes citrus stubborn disease and vegetative disorders in Apiaceae, such as carrot and celery, and in other hosts. Detection methods include indexing in indicator plants, in vitro culturing in specific media, and serological and molecular amplifi...

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Autores principales: Alfaro-Fernández, Ana, Ibanez, Isabel, Bertolini, Edson, Hernandez-Llopis, D., Cambra, Mariano, Font-San-Ambrosio, Isabel
Formato: Artículo
Lenguaje:Inglés
Publicado: 2018
Acceso en línea:http://hdl.handle.net/20.500.11939/6016
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author Alfaro-Fernández, Ana
Ibanez, Isabel
Bertolini, Edson
Hernandez-Llopis, D.
Cambra, Mariano
Font-San-Ambrosio, Isabel
author_browse Alfaro-Fernández, Ana
Bertolini, Edson
Cambra, Mariano
Font-San-Ambrosio, Isabel
Hernandez-Llopis, D.
Ibanez, Isabel
author_facet Alfaro-Fernández, Ana
Ibanez, Isabel
Bertolini, Edson
Hernandez-Llopis, D.
Cambra, Mariano
Font-San-Ambrosio, Isabel
author_sort Alfaro-Fernández, Ana
collection ReDivia
description Spiroplasma citri is a phloem-limited bacterium that causes citrus stubborn disease and vegetative disorders in Apiaceae, such as carrot and celery, and in other hosts. Detection methods include indexing in indicator plants, in vitro culturing in specific media, and serological and molecular amplification methods, e.g., conventional and real-time polymerase chain reaction (PCR). A protocol for the detection of S. citri in carrot seeds, roots and leaves by real-time PCR with a TaqMan probe was developed and assayed. The protocol was specific and sensitivity was 102 times higher than conventional PCR for S. citri detection. One and two samples out of 20, which consisted of five whole seeds per sample, were positive for S. citri by real-time PCR, which implies an infection rate of at least 1-2%. Seed transmission was assessed when analysing the roots of the seedlings grown in an insect-free containment growth chamber (P2 level) 240 days post-sowing. Transmission rates of 30.1% and 37.6% were estimated. The symptoms observed in all the seedlings that tested positive for the pathogen included twisted petioles, filiform, mosaic, yellowing or red leaves, and protuberances, deformation or branching in roots. The epidemiological consequences of S. citri transmission in carrot seeds are discussed. The role of S. citri inoculum in carrot crops cannot be ruled out at increasing infection rates in other hosts. The control of carrot seed lots for this quarantine pathogen must be undertaken.
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institution Instituto Valenciano de Investigaciones Agrarias (IVIA)
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spelling ReDivia60162025-04-25T14:45:54Z Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection Alfaro-Fernández, Ana Ibanez, Isabel Bertolini, Edson Hernandez-Llopis, D. Cambra, Mariano Font-San-Ambrosio, Isabel Spiroplasma citri is a phloem-limited bacterium that causes citrus stubborn disease and vegetative disorders in Apiaceae, such as carrot and celery, and in other hosts. Detection methods include indexing in indicator plants, in vitro culturing in specific media, and serological and molecular amplification methods, e.g., conventional and real-time polymerase chain reaction (PCR). A protocol for the detection of S. citri in carrot seeds, roots and leaves by real-time PCR with a TaqMan probe was developed and assayed. The protocol was specific and sensitivity was 102 times higher than conventional PCR for S. citri detection. One and two samples out of 20, which consisted of five whole seeds per sample, were positive for S. citri by real-time PCR, which implies an infection rate of at least 1-2%. Seed transmission was assessed when analysing the roots of the seedlings grown in an insect-free containment growth chamber (P2 level) 240 days post-sowing. Transmission rates of 30.1% and 37.6% were estimated. The symptoms observed in all the seedlings that tested positive for the pathogen included twisted petioles, filiform, mosaic, yellowing or red leaves, and protuberances, deformation or branching in roots. The epidemiological consequences of S. citri transmission in carrot seeds are discussed. The role of S. citri inoculum in carrot crops cannot be ruled out at increasing infection rates in other hosts. The control of carrot seed lots for this quarantine pathogen must be undertaken. 2018-05-09T16:30:55Z 2018-05-09T16:30:55Z 2017 article Alfaro-Fernandez, A., Ibanez, I., Bertolini, E., Hernandez-Llopis, D., Cambra, M., Font, M. I. (2017a). Transmission of spiroplasma citri in carrot seeds and development of a real-time pcr for its detection. Journal of Plant Pathology, 99(2), 371-379. 1125-4653 http://hdl.handle.net/20.500.11939/6016 10.4454/jpp.v99i2.3878 en Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ electronico
spellingShingle Alfaro-Fernández, Ana
Ibanez, Isabel
Bertolini, Edson
Hernandez-Llopis, D.
Cambra, Mariano
Font-San-Ambrosio, Isabel
Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection
title Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection
title_full Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection
title_fullStr Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection
title_full_unstemmed Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection
title_short Transmission of Spiroplasma Citri in Carrot Seeds and Development of a Real-Time Pcr for its Detection
title_sort transmission of spiroplasma citri in carrot seeds and development of a real time pcr for its detection
url http://hdl.handle.net/20.500.11939/6016
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