Potato leafroll virus detection by RT-PCR in field-collected aphids

Singh, R. P.; Kurz, Jolanta; Boiteau, Gilles; Moore, L. M.

doi:10.1007/bf02851574

Cited by 21 publications

(6 citation statements)

References 13 publications

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“…Conventional polymerase chain reaction (PCR) or reverse transcription-PCR (RT-PCR) which have high sensitivity and specificity over ELISA-based methods and have been used to detect DNA viruses or RNA viruses, respectively (Bostan and Peker, 2009;Stark et al, 2008;Zaghloul, 2011). RT-PCR and immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) have been employed for detection of PLRV in plant or in aphid vectors (Ahouee et al, 2010;Hadidi et al, 1993;Peiman and Xie, 2006;Singh et al, 1995;1997). Notomi et al (2000) developed loop-mediated isothermal amplification (LAMP) to amplify DNA with high specificity.…”

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confidence: 99%

“…RT-PCR (Hadidi et al, 1993;Peiman and Xie, 2006;Singh et al, 1995;1997) and IC-RT-PCR (Ahouee et al, 2010) have been developed for the rapid and accurate detection of PLRV in plants or/and aphids. Although these conventional PCR-based methods increased rapidness and accuracy for PLRV detection, these methods require expensive and sophisticated instruments for the PLRV amplification and detection of the PCR products.…”

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju¹

2011

The Plant Pathology Journal

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A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RT-PCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju¹

2011

The Plant Pathology Journal

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“…RT-PCR is an extremely sensitive technique and has been used widely to detect plant viruses that occur at relatively low concentrations in their host plants or that are unevenly distributed in the plants (16). It also has been used to detect several persistently and nonpersistently transmitted viruses in single aphids (1,20,(26)(27)(28)(29)(30)32,37). All these studies emphasized that virus detection in aphids by RT-PCR has a potential application in monitoring virus incidence in the fields.…”

Section: Discussionmentioning

confidence: 99%

“…The reverse-transcription polymerase chain reaction (RT-PCR) methods have been widely used for the detection of RNA plant viruses that occur in low concentrations. Previous studies have shown that viruses can be detected in individual viruliferous aphids using RT-PCR (1,20,(26)(27)(28)(29)(30)32,37). In addition to evaluating the transmission efficiency of several different plant viruses by A. glycines, the objectives of our study included the development of an RT-PCR assay for detecting SbDV in single viruliferous aphids as a means for predicting possible incidence of SbDV in soybean fields.…”

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Aphis glycines as a Vector of Persistently and Nonpersistently Transmitted Viruses and Potential Risks for Soybean and Other Crops

et al. 2006

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The recently introduced soybean aphid (Aphis glycines), which is widespread in the soybean-growing regions in the United States, is the only aphid able to develop large colonies on soybean. Although its potential as a vector of plant viruses is recognized, reports on virus transmission efficiency by this aphid species are limited. In the present study, we examined the ability of A. glycines to transmit several economically important viruses. The results showed that A. glycines transmitted the potyviruses Bean yellow mosaic virus (BYMV) and Soybean mosaic virus from soybean to soybean more efficiently than Myzus persicae. However, M. persicae transmitted the alfamovirus Alfalfa mosaic virus and the potyviruses Tobacco etch virus (TEV) and Tobacco vein mottling virus (TVMV) from tobacco to tobacco more efficiently than A. glycines. This is the first report to demonstrate that the soybean aphid can vector TEV and TVMV, two economically important tobacco viruses. This also is the first report to document successful transmission of BYMV by A. glycines. All attempts to transmit the nepovirus Tobacco ringspot virus by A. glycines were unsuccessful, regardless of the length of the acquisition and inoculation feeding periods. Although the luteovirus Soybean dwarf virus (SbDV) was widely distributed in red and white clover in Kentucky, it was not detected in soybean. All transmission experiments of SbDV by A. glycines were unsuccessful. A reverse-transcription polymerase chain reaction (RT-PCR) assay was developed to detect SbDV in single aphids using a pair of primers designed to amplify a 372-bp PCR fragment in the coding region of SbDV coat protein. Although A. glycines was not a vector of SbDV, the virus was detected in 100% of tested aphids by RT-PCR after a 24- to 48-h virus acquisition access feeding. The practical applications of RT-PCR in detecting persistently transmitted viruses are discussed.

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“…Sample extraction from sticky traps using alkane solvents has been found in other entomological studies to preserve the integrity of samples for identification (Davidson et al, 2015) and of viral RNA for assay (Boonham et al, 2002;Congdon et al, 2019). Singh et al, 1997, found detectability of viral RNA by RT-PCR to be retained in individual aphids stored for up to seven years in 70% ethanol.…”

Section: Introductionmentioning

confidence: 99%

Can a PCR assay of aphids caught in‐crop on yellow sticky traps inform field level barley yellow dwarf virus risk assessment?

Bates

Pope

Holland

2020

Annals of Applied Biology

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Infection with barley yellow dwarf virus (BYDV), caused by strains of virus belonging to the 5 family Luteovirideae including BYDV-PAV, can result in significant yield losses in autumn sown 6 cereals following transmission by aphid vectors such as Rhopalosiphum padi (Homoptera: 7Aphididae) and Sitobion avenae (Homoptera: Aphididae). Spatial and temporal variance in the 8 infectivity of alate populations influences risk to crops from the disease, which is greatest on infection at early crop growth stages. A decision support system (DSS) to guide optimised integration of crop protection strategies through risk assessment would help avoid unnecessary application of synthetic insecticides. This study contributes to the development of a DSS by exploring the viability and relevance of a methodology to detect virus levels in individual aphids trapped in-crop using yellow sticky traps. Using a reverse transcription polymerase chain reaction (RT-PCR) assay, the detectability of virus from a BYDV-PAVpositive control colony was found not to be reduced by the process of trapping, extraction and cold storage, but did drop significantly after between three and seven days of exposure on trap. This method has potential to contribute to localised risk assessment and guide optimisation of crop protection strategies.

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Potato leafroll virus detection by RT-PCR in field-collected aphids

Cited by 21 publications

References 13 publications

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Aphis glycines as a Vector of Persistently and Nonpersistently Transmitted Viruses and Potential Risks for Soybean and Other Crops

Can a PCR assay of aphids caught in‐crop on yellow sticky traps inform field level barley yellow dwarf virus risk assessment?

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