Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR

Bertolini, Edson; Moreno, Aránzazu; Capote, Nieves; Olmos, Antonio; Luis, Ana de; Vidal, Eduardo; Pérez-Panadés, Jordi; Cambra, M.

doi:10.1007/s10658-007-9206-9

Cited by 90 publications

(68 citation statements)

References 33 publications

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“…With the exception of the positive controls, all samples tested negative for PVX, PYVV, PVS and PVV (Table 1). The RT-qPCR results match well with the NGS data, validating its potential as the molecular diagnostic tool of choice in the asymptomatic detection of known viruses, since it has been estimated that RT-qPCR is about 10 3 -10 5 times more sensitive than either ELISA or conventional RT-PCR (Bertolini, Moreno, Capote, Olmos, de Luis, Vidal, Pérez-Panadés & Cambra, 2008;Medina, Gutiérrez & Marín, 2015).…”

Section: Discussionsupporting

confidence: 62%

Detection and sequencing of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in a volunteer plant of Solanum tuberosum L. cv. Diacol-Capiro

2017

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Viral diseases are among the most limiting factors in the production of potato in Colombia and the rest of the world. The best strategy to control plant viruses consists on the use of certified seed tubers, control of arthropod vectors and the use of adequate crop management practices that reduce mechanical transmission and the presence of viral reservoirs like weeds and volunteer plants. However, the successful implementation of these practices relies on the availability of highly sensitive techniques that allow for the asymptomatic detection of viruses. In this work, we tested the performance of Next-generation sequencing (NGS) and real time RT-PCR (RT-qPCR) on a single volunteer potato plant (cv. Diacol-Capiro) growing naturally in a seed-tuber storage facility in Yarumal (Antioquia). Our NGS results demonstrate a mixed infection with Potato virus Y (PVY) and Potato leafroll virus (PLRV). RT-qPCR was performed in roots, main stolons, crown (root collar) and upper, middle and lower leaves using specific primers for PVY, PLRV, Potato virus S (PVS), Potato virus V (PVV), Potato virus X (PVX) and Potato yellow vein virus (PYVV). Only PVY and PLRV were detected in good agreement with the NGS data. This work demonstrates the usefulness of both techniques for supporting integrated management of plant viruses in potato, including virus detection in natural reservoirs such as volunteer plants and weeds.

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Section: Discussionsupporting

confidence: 62%

Detection and sequencing of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in a volunteer plant of Solanum tuberosum L. cv. Diacol-Capiro

2017

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“…The universal primers Pin 1 and Pin 2 (Bertolini et al, 2008), the T36CP primers (Hilf and Garnsey, 2000) and those designed by Bertolini et al (2008) for real-time PCR were capable of recognizing 100% of the Chilean CTV isolates used in this study.…”

Section: Molecular Characterizationmentioning

confidence: 95%

Aggressive Citrus tristeza virus isolates in Chile are MCA13-positive and VT type, while mild isolates are MCA13-negative and T30 type

et al. 2015

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Citrus tristeza virus (CTV) was reported in the 1960's to affect Meyer lemon trees in Chile, but no field symptoms were observed. This study performed a complete biological, serological and molecular characterization of one hundred CTV isolates obtained from different hosts and geographical origins. Decline symptoms (DI) were not found on trees grafted onto sour orange. In Pica Oasis in northern Chile, stem pitting (SP) was found to affect grapefruit and Mexican lime trees. Most isolates present in the central area were considered mild (MCA-13-negative), while in the northern area aggressive isolates were observed and detected. Some of these isolates were capable of causing SP on grapefruit under field and greenhouse conditions and on sweet orange under greenhouse conditions. Almost all of these isolates were MCA13-positive and had nucleotide sequences associated with the VT genotype.

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“…Same procedure was used to estimate the number of CTV-viruliferous aphids in Field 2. The squashed aphid individuals (Figure 1) from both plots were analysed using the squash real-time RT-PCR procedure (Bertolini et al, 2008).…”

Section: Aphid Species Monitoring and Estimation Of The Number Of Ctvmentioning

confidence: 99%

Epidemiology of Citrus tristeza virus in nursery blocks of Citrus macrophylla and evaluation of control measures

Vidal

Moreno

Bertolini

et al. 2012

Span. j. agric. res.

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The control of Citrus tristeza virus (CTV), the causal agent of tristeza disease, is essential to guarantee a productive citrus industry. The specific features of a nursery block make the control of the natural spread of viruses transmitted by vectors more difficult. Thus, the knowledge of the epidemiology of CTV in nursery blocks is basic to the design control strategies of the disease. Two experimental nursery plots of alemow (Citrus macrophylla) were planted in open field in Moncada (Valencia, Spain) (with a high CTV prevalence, 85%), and in Alcanar (Tarragona, Spain) with 700 and 843 nursery plants, respectively. The plants were analysed by tissue print-ELISA using 3DF1 and 3CA5 CTVspecific monoclonal antibodies. The CTV prevalence estimated after one year of cultivation in both plots was of 24.37% and 3.91%, respectively. At the same time, a commercial nursery plot grown under plastic-net covers established in Alcanar with 1,200 plants was also analysed as above, being the CTV prevalence estimated of 0.00%. The aphid activity present in the nursery plots was monitored. Aphis spiraecola was the most abundant aphid species visiting the plants in the nursery plots grown in the open field, whereas the aphid activity registered in the plants grown under the plasticnet covers was practically zero. The percentage of individual aphids carrying CTV PCR-amplifiable targets detected by squash real-time RT-PCR in the open-field nursery plots was 17.5% in Moncada and 1.67% in Alcanar. No significant differences in the CTV prevalence between treated and non-treated plants with horticultural mineral oils were found in the area with the high CTV-inoculum pressure.Additional key words: alemow; horticultural mineral oil treatments; physical barriers; semi-persistent virus; squash real-time RT-PCR; tissue print-ELISA; vector intensity. Resumen Epidemiología del virus de la tristeza de los cítricos en bloques de vivero de Citrus macrophylla y evaluación de medidas de controlEl control del virus de la tristeza de los cítricos (Citrus tristeza virus, CTV), agente causal de la enfermedad de la tristeza, es esencial para garantizar una industria citrícola productiva. Las características específicas de los viveros hacen más difícil el control de los virus de vegetales transmitidos por insectos vectores. Así, el conocimiento de la epidemiología de CTV en bloques de plantas de vivero es básico para diseñar estrategias de control de la enfermedad. Dos parcelas experimentales de vivero cultivadas en campo abierto de Citrus macrophylla se establecieron en Moncada (Valencia, España) (con una alta prevalencia de CTV, 85%) y Alcanar (Tarragona, España) con 700 y 843 plantas de vivero, respectivamente. Las plantas se analizaron mediante Inmunoimpresión-ELISA usando los anticuerpos monoclonales específicos 3DF1 y 3CA5 de CTV. La prevalencia de CTV estimada en ambas parcelas después de un año de cultivo fue del 24,37% y del 3,91%, respectivamente. Al mismo tiempo, una parcela de vivero comercial cultivada bajo cubierta de plástico con...

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Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR

Cited by 90 publications

References 33 publications

Detection and sequencing of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in a volunteer plant of Solanum tuberosum L. cv. Diacol-Capiro

Detection and sequencing of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in a volunteer plant of Solanum tuberosum L. cv. Diacol-Capiro

Aggressive Citrus tristeza virus isolates in Chile are MCA13-positive and VT type, while mild isolates are MCA13-negative and T30 type

Epidemiology of Citrus tristeza virus in nursery blocks of Citrus macrophylla and evaluation of control measures

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