Spread of Grapevine Leafroll-Associated Virus 3 (GLRaV-3) by Scale Insects


Project Number
AD 936

Project Title
Spread of grapevine leafroll-associated virus 3 (GLRaV-3) by scale insects

Project Leader
Krüger, K

Institution
University of Pretoria

Team Members
Mawela, K V
Saccaggi, D

Project Description
A transmission protocol – assay, which is required as support technology for the development of virus-resistant grapevines has been developed based on the transmission characteristics of Planococcus ficus. The transmission experiments have demonstrated that an acquisition access period (AAP) of 24 hours is sufficient for P. ficus to acquire the virus, and not more than 24 hours as stated in the literature. The experiments have further shown that an inoculation access period (IAP) of 24 hours or less is enough for the vector to inoculate healthy grapevine plants. Experiments on the persistence of the virus in P. ficus indicate that the virus persists for at least 24 hours in the vector when feeding on healthy grapevine plants.

The present study is the first to develop a method to detect GLRaV in individual mealybugs, including first-instar nymphs. This result is of great importance as it is now possible directly to determine the percentage of mealybugs or the percentage of different developmental stages in a population that carry the virus.

Trials are under way to determine alternative vectors, their transmission biology and life cycle in controlled laboratory experiments. A survey to determine alternative host plants of vectors is ongoing.

The project has been expanded to examine the movement of mealybugs in the field to determine their role in the secondary spread of the disease. In addition, the transmission efficiency of GLRaV-3 in the presence and absence of GVA (grapevine virus A) will be evaluated.

Presentation(s)

Kruger, K. 2001. Spread of grapevine leafroll-associated virus 3 (GLRaV-3) by scale insects. Presentation at the 1st Grapevine Virus Workshop. May, Simondium, South Africa.

Kruger, K. 2002. Spread of grapevine leafroll-associated virus 3 (GLRaV-3) by scale insects. Presentation at the 2nd Grapevine Virus Workshop. May, Simondium, South Africa.

Kruger, K. 2003. Spread of grapevine leafroll-associated virus 3 (GLRaV-3) by scale insects (Hemiptera: Coccoidea). Presentation at the 3rd Grapevine Virus Workshop. May, Simondium, South Africa.

Kruger, K. 2003. Transmission of grapevine leafroll-associated virus 3 (GLRaV-3) by Planococcus ficus (Hemiptera: Pseudococcidae). Paper presented at the 14th Entomological Congress of the Entomological Society of Southern Africa. 6-9 July, Pretoria, South Africa.

Kruger, K. 2004. The role of insects vectors in the transmission of grapevine leafroll disease. Presentation at the 4th Grapevine Virus Workshop, May, Simondium, South Africa.

Kruger, K. 2005. The role of insects vectors in the transmission of grapevine leafroll disease. Presentation at the 5th Grapevine Virus Workshop. May, Simondium, South Africa

Mawela, K V, Douglas, N, Van der Merwe, J and Kruger, K. 2006. Temperature dependent development of the longtailed mealybug Pseudococcus longispinus Targioni-Tozzetti (Hemiptera: Pseudococcidae) on grapevine. Paper presented at the 15th Entomological Congress of the Entomological Society of Southern Africa. 10-13 July, Grahamstown, South Africa.

Saccaggi, D L, Kruger, K, Pietersen, G and MacDonald, C. 2006. A PCR-based method for identification of three mealybug species (Hemiptera: Pseudococcidae). Paper presented at the 15th Entomological Congress of the Entomological Society of Southern Africa. 10-13 July, Grahamstown, South Africa.

Douglas, N and Kruger, K. 2005. Persistence of grapevine leafroll-associated virus 3 (GLRaV-3) in Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae).

Kruger, K. 2006. Spread of GLRaV-3 by scale insects (Coccoide). Presentation at the 6th Winetech Grapevine Virus Workshop. May, Simondium, South Africa.

Saccaggi, D L, Kruger, K and Pietersen, G. 2006. Rapid identification of three mealybug species by multiplex PCR. Paper presented at the 15th Meeting of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 3-7 April, Stellenbosch, South Africa.

Kruger, K, Douglas, N and Saccaggi. 2006. Grapevine leafroll-associated virus 3 – vector interactions: Transmission by the mealybugs Planococcus ficus and pseudococcus longispinus (Hemiptera: Pseudococcidae). Paper presented at the 15th Meeting of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 3-7 April, Stellenbosch, South Africa.

Douglas, N and Kruger, K. 2006. Grapevine leafroll-associated virus 3 transmission efficiency of Planococcus ficus and Pseudococcus longispinus (Hemiptera: Pseudococcidae). Paper presented at the 15th Meeting of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 3-7 April, Stellenbosch, South Africa.

Kruger, K, Douglas, N and Ferreira, M. 2007. Grapevine leafroll disease, mealybugs and vineyards. Presentation at the 7th Grapevine Virus Workshop. May, Stellenbosch, South Africa.

Douglas, N, Malherbe, G P and Kruger, K. 2008. Grapevine leafroll-associated virus 3 (GLRaV-3) extraction from grapevines and mealybugs (Hemiptera: Pseudococcidae) for quantitative real-time PCR. Paper presented at the 23rd International Congress of Entomology. 6-12 July, Durban, South Africa.

Ferreira, M, Pietersen, G, Barros, E and Kruger, K. 2008. Determining dispersal patterns of Planococcus ficus (Hemiptera: Pseudococcidae) among South African vineyards using RAPD markers. Paper presented at the 23rd International Congress of Entomology. 6-12 July, Durban, South Africa.

Kruger, K and Douglas, N. 2008. Grapevine leafroll virus extraction from grapevines and mealybugs for quantitative real-time PCR. Presentation at the 8th Grapevine Virus Workshop. August, Stellenbosch, South Africa.

Ferreira, M, Barros, E, Pietersen, G and Kruger, K. 2008. Secondary spread of GLRaV-3 through the vine mealybug Planococcus ficus. Presentation at the 8th Grapevine Virus Workshop. August, Stellenbosch, South Africa.

Ferreira, M, Pietersen, G, Barros, E and Krüger, K. 2009. Development of a RAPD protocol to determine dispersal patterns of Planococcus ficus (Hemiptera: Pseudococcidae) among South African vineyards using RAPD markers. Bioinformatics and genomics approaches for functional gene expression in vector-virus-host interactions. Paper presented at the Joint Symposium and University of the Witwatersrand and University of Arizona. March, Johannesburg, South Africa.

Kruger, K. 2010 Control of leafroll disease through vector control. Paper presented at the 4th International Congress of the South African Society for Enology and Viticulture. 8-10 July, Cape Town, South Africa.

Krüger, K and Douglas, N. 2009. Transmission of grapevine leafroll-associated virus 3 (GLRaV-3) by three soft scale insect species (Hemiptera: Coccidae) and notes on their developmental biology on grapevine. Paper presented at the 16th Congress of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 31 August – 4 September 2009, Dijon, France.

Douglas, N, Pietersen, G and Krüger, K. 2009. A real-time RT-PCR assay for the detection and quantification of grapevine leafroll-associated virus 3 (GLRaV-3) in Vitis vinifera L. (Vitaceae) and Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae).
Paper presented at the 16th Congress of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 31 August – 4 September 2009, Dijon, France.

Ferreira, M, Barros, E, Pietersen, G and Kruger, K. 2009. Determining dispersal patterns of Planococcus ficus (Hemiptera: pseudococcidae) among South African vineyards using RAPD markers. Presentation at the 9th Grapevine Virus Workshop. September, Stellenbosch, South Africa.

Article
Douglas, N, Kruger, K. 2008. Transmission efficiency of Grapevine leafroll-associated virus 3 (GLRaV-3) by the mealybugs (Planococcus ficus and Pseudococcus longispinus) European Journal of Plant Pathology, Mnth Oct v. 122 (2) (p. 207-212)

Saccaggi, D L, Kruger, K, Pietersen, G. 2008. A multiplex PCR assay for the simultaneous identification of three mealybug species (Hemiptera: Pseudococcidae) Bulletin of Entomological Research, Mnth Feb v. 98 (1) (p. 27-33)

Walton, V M, Kruger, K, Saccaggi, D L, Millar, I M. 2009. A survey of scale insects (Sternorryncha: Coccoidea) occurring on table grapes in South Africa, Journal of Insect Science, v. 9 (47) (p. 1-6)

Paper
Kruger, K. 2003. Transmission of grapevine leafroll-associated virus 3 (GLRaV-3) by Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) IN: Stals, R (ed). Proceedings of the 14th Congress of the Entomological Society of Southern Africa. 6-9 July 2003, Pretoria, South Africa. (p. 46-47) Entomological Society of Southern Africa, Pretoria, South Africa.

Douglas, N, Kruger, K. 2005. Grapevine leafroll-associated virus 3 transmission efficiency of Planococcus ficus and Pseudococcus longispinus (Hemiptera: Pseudococcidae) IN: Stals, R (ed). Proceedings of the 15th Entomological Congress of the Entomological Society of Southern Africa. 10-13 July 2005, Grahamstown, South Africa. (p. 191-192)

Douglas, N, Kruger, K. 2005. Persistence of grapevine leafroll-associated virus 3 in Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) IN: Stals, R (ed). Proceedings of the 15th Entomological Congress of the Entomological Society of Southern Africa. 10-13 July 2005, Grahamstown, South Africa. (p. 19) Entomological Society of Southern Africa, Pretoria, South Africa.

Mawela, K V, Douglas, N, Van der Merwe, J, Kruger, K. 2005. Temperature dependent development of the longtailed mealybug Pseudococcus longispinus Targioni-Tozzetti (Hemiptera: Pseudococcidae) on grapevine, IN: Stals, R (ed). Proceedings of the 15th Congress of the Entomological Society of Southern Africa. 10-13 July 2005, Grahamstown, South Africa. (p. 73)

Saccaggi, D L, Kruger, K, Pietersen, G, MacDonald, C. Stals, R (ed) (ed). 2005. A PCR-based method for the identification of three mealybug species (Hemiptera: Pseudococidae) IN: Proceedings of the 15th Congress of the Entomological Society of Southern Africa. 10-13 July 2005, Grahamstown, South Africa. (p. 51)

Kruger, K, Saccaggi, D L, Douglas, N. 2006. Grapevine leafroll-associated virus 3 vector interactions: Transmission by the mealybug Planococcus ficus and Pseudococcus longispinus (Hemiptera: Pseudococcidae) IN: Proceedings of the 16th Congress of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 3-7 April 2006, Stellenbosch, South Africa. (p. 130-131)

Saccaggi, D L, Kruger, K, Pietersen, G. 2006. Rapid identification of three mealybug species by multiplex PCR, IN: Proceedings of the 16th Congress of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 3-7 April 2006, Stellenbosch, South Africa. (p. 124-125)

Thesis
Mawela, K V. 2005. Temperature requirements of the longtailed mealybugs Pseudococcus longispinus (Targionie Tozzetti) Hemiptera: Pseudococcidae) on grapevine. MiniAgric Thesis. University of Pretoria, Pretoria, South Africa.

Saccaggi, D L. 2006. Development of molecular techniques to identify mealybugs (Hemiptera: Pseudoccidae) of importance on grapevines in South Africa. MSc. Universtity of Pretoria, Pretoria, South Africa.

Smit, N. 2008. A real time RT-PCR assay for the detection and quantification of grapevine leafroll-associated virus 3 in Vitis vinifera (Vitaceae) and Planococcus ficus (Signoret) Hemiptera: Pseudococcidae. MSc. University of Pretoria, Pretoria, South Africa.

FinalReport.pdf

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