The Cloning and Molecular Characterisation of the Coat Protein Gene of a South African Isolate of a Grapevine Leafroll-Associated Virus 3


Project Number
IWBT 04-05

Project title
The cloning and molecular characterisation of the coat protein gene of a South African isolate of a grapevine leafroll-associated virus 3

Project leader
Burger, J T

Institution
University of Stellenbosch. Faculty of AgriSciences. Institute for Wine Biotechnology

Team members
Bellstedt, D
Maree, H J
Vivier, M A

Project description
We aim to introduce virus resistance in grapevine cultivars by genetic engineering. The existing project targets a specific virus; grapevine leafroll-associated virus 3 (GLRaV-3), which is the most destructive virus infecting grapevines in South Africa. Our approach is to clone and characterise the coat protein gene of an South African isolate of GLRaV-3. Clones will be genetically engineered for subsequent expression in grapevines, as a possible resistance mechanism to this virus. Most virus diseases in grapevine seem to be caused by complexes of viruses. We wish to expand our efforts to include a broad-spectrum virus resistance approach in grapevines. We propose to do this by expressing a ribosome inactivating protein in transgenic grapevines.

This project comprises a first attempt to introduce pathogen-derived resistance against grapevine leafroll disease in Vitis vinifera cultivars. So far, we have isolated and characterised the coat protein gene of the viral pathogen, causing this disease. We have constructed vectors to express the GLRaV-3 CP-gene on a transcriptional and translational level in plants, and used these to transform tobacco and grapevine. In a second approach, we propose to introduce broad-spectrum virus resistance in grapevine by expressing a ribosome inactivating protein gene, from an unrelated plant, in grapevine. A few candidate genes have been isolated and cloned into different expression systems. These are in the process of being evaluated for expression efficiency.

Presentation(s)
Van Straten, C D and Burger, J T. 1998. The isolation and characterisation of the coat protein of a South African isolate of GLRaV-3. 22nd National Congress of the South African Society for Enology and Viticulture, Somerset West, South Africa.

Wilsen, K L and Burger, J T. 1998. Screening the grapevine genome for an RIP gene. 22nd National Congress of the South African Society for Enology and Viticulture, Somerset West, South Africa.

Burger, J T, Van Straten, C D and Wilsen, K L. 1999. Grapevine viral disease: can we control them? 23rd National Congress of the South African Society for Enology and Viticulture, Somerset West, South Africa.

Van Straten, C D and Burger, J T. 1999. Cloning the coat protein gene of a South African isolate of GLRaV-3 in a plant expression vector. 37th Congress of the South African Society for Plant Pathology, Pietermaritzburg, South Africa.

Wilsen, K L and Burger, J T. 1999. Investigating the introduction of a non-specific antiviral mechanism into grapevine. 37th Congress of the South African Society for Plant Pathology, Pietermaritzburg, South Africa.

Burger, J T, Graf, J A and Van Straten, C D. 2000. Construction of pDVC+. A plant expression vector bearing GFLV and GLRaV-3 coat protein genes. BioY2K Combined Millennium Meeting. Grahamstown. South Africa.

Burger, J T, Van Straten, C D, Vivier, M A and, Pretorius I S. 2000. Progress in the establishment of grapevine leafroll disease resistant grape varieties. 2nd International Congress of the South African Society for Viticulture and Enology. Cape Town. South Africa.

Burger, J T and Wilsen, K L. 2000. Towards the introduction of a broad-spectrum antiviral mechanism into grapevine. 13th Meeting of the International Council for the Study of Viruses of Grapevine, Adelaide, Australia.

Maree, H J and Burger, J T. 2002. Expression of Dianthin, a ribosome inactivating protein for broad-spectrum virus resistance. 40th Congress of the South African Society for Plant Pathology, Dikhololo, South Africa.

Article
Burger, J. 1999. Rolblaarsiekte: ‘n nuwe beheerstrategie? Wynboer, Mnth Dec (p. T4-5)

Thesis
Van Straten, C D. 2000. The construction of plant expression for the introduction of leafroll disease resistance in grapevine. MSc. University of Stellenbosch, Stellenbosch.

Wilsen, K L. 2000. Investigating the introduction of a broad-spectrum antiviral mechanism into grapevine. MSc. University of Stellenbosch, Stellenbosch.

Maree, H J. 2002. Molecular characterisation of ribosome inactivating protein-expression clones. MSc (Wine Biotechnology) University of Stellenbosch, Stellenbosch.

FinalReport.pdf

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