Development of a Large-Scale Detection Method for Grapevine Leafroll-Associated Virus 3


Project Number
PPRI GP 02

Project title
Development of a large-scale detection method for grapevine leafroll-associated virus 3

Project leader
Pietersen, G

Institution
ARC Plant Protection Research Institute

Project description
Grapevine leafroll disease is a serious problem for the South African wine industry but in recent years a successful control strategy based on rouging of infected vines, control of mealybug numbers and dispersal has been developed. Rouging can be effectively achieved in red cultivars by visual vine-for-vine detection annually in late autumn. Visual assessment of leafroll infection can however not be done with white cultivars as leafroll symptoms are not obvious in these and to achieve control of leafroll vines must be subjected to a virus detection method annually prior to rouging. Currently the sensitive techniques, PCR or real-time PCR, which allow combining of large numbers of vines for tests (to make the tests cheaper) are too expensive, because of their infrastructure requirements, to be done on the scale required to achieve yearly testing of individual white cultivar vines. Furthermore these tests must be conducted in laboratories with relatively well trained personnel. We propose a project to develop and assess the ability of relatively new nucleic acid isothermal amplification techniques called loop mediated isothermal amplification and cycleave isothermal and chimeric primer-initiated amplification (Cycleave ICAN) for the detection of grapevine leafroll-associated virus 3 the cause of leafroll disease in South Africa. The techniques will be adapted to do large-scale decentralized detection of GLRaV-3, in order to identify individually infected white cultivar vines to allow their ultimate removal in order to control the disease. Should one or both of these techniques work well the project will be expanded by two years to develop similar techniques for grapevine virus A and B (which currently are tested by ELISA using expensive important reagents.

Presentation(s)
Walsh, H A and Pietersen, G. 2010. A novel method for the detection of Candidatus Liberibacter africanus using isothermal and chimeric primer-initiated amplification of nucleic acid. Paper presented at the 6th Citrus Research Symposium. 15-18 August, Drakensberg, South Africa.

Walsh, H A and Pietersen, G. 2010. Novel methods for the detection of grapevine leafroll-associated virus type 3 and Candidatus Liberibacter africanus using isothermal amplification techniques. Presentation at the ARC Plant Protection Research Institute Science Day. November, Pretoria, South Africa.

Pietersen, G and Walsh, H.A. 2011. Further progress towards grapevine leafroll control in South Africa 2010 – 2011. Paper presented at 11th Winetech Grapevine Virus Workshop. May 2011, ARC Infruitec-Nietvoorbij, Stellenbosch, South Africa.

Walsh, H A and Pietersen, G. 2011. Detecting grapevine leafroll-associated virus-3 using loop-mediated isothermal amplification (LAMP) and isothermal and chimeric primer-initiated amplification of nucleic acid (ICAN). Paper presented at the FABI Information Seminar. 31 March

Walsh, H A and Pietersen, G. 2011. Detecting grapevine leafroll-associated virus-3 using loop-mediated isothermal amplification (LAMP) and isothermal and chimeric primer-initiated amplification of nucleic acid (ICAN). Lecture. University of Pretoria, Department of Microbiology and Plant Pathology. 15 May, Pretoria, South Africa.

Walsh, H A and Pietersen, G. 2013. Detecting grapevine leafroll-associated virus type 3 using loop-mediated amplification. Paper presented at the 18th Conference of the South African Society for Microbiology. 24-27 November, Warmbaths, South Africa.

Pietersen, G and Walsh, H A. 2012. Development of a LAMP technique for the control of grapevine leafroll associated virus 3 (GLRaV3) in infected white cultivar vines by rogueing. Paper presented at the 17th International Council for the Study of Virus and Virus-like Diseases of the Grapevine Conference. 7-14 October, Davis, California, United States.

Article
Walsh, H A, Pietersen, G. 2013. Rapid detection of grapevine leafroll-associated virus 3 using a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method, Journal of Virological Methods, Mnth 194 (p. 308-316)

FinalReport.pdf

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