Optimisation of a quantitative RT-PCR assay for the reproducible detection of grapevine viruses in rootstock material
Burger, J T
University of Stellenbosch. Department of Genetics
Burger, J T
Freeborough, M J
The supply of virus free grapevine material for planting is one of the most effective ways of controlling the spread of viral disease in South Africa. To achieve this end, plat material is subject to regulations and control by the Vine Improvement Association (VIA). All plant material passing through this scheme needs to be tested for virus infection. A major challenge to this scheme is the detection of viruses in rootstock material, which have proved difficult for detection assays in the past. This project plans to focus on establishing and validating a sensitive and reliable detection assay of GLRaV-3 and GVA using a SYBR Green quantitative (real-time) reverse transcription polymerase chain reaction (SG-qRT-PCR).
Objectives for the project are:
1. Identification and establishment of reliable sources of GLRaV-3 and GVA infected rootstock material.
2. Design GLRaV-3 and GVA specific qRT-PCR primer sets
3. Optimisation of sample extraction protocols
4. Optimisation of SG-qRT-PCR for both GLRaV-3 and GVA in infected rootstock material.
5. Trial detection in the final year.