Project title
Development of ELISA for simultaneous detection of grapevine virus A (GVA) and grapevine virus B (GVB)
Project leader
Goszczynski, D E
Institution
ARC Plant Protection Research Institute
Team members
Jooste, A E C
Goszczynski, T
Project description
Rugose wood complex of grapevines comprising four graft transmissible diseases: Kober stem grooving (KSG), Corky bark (CB), LN33 stem grooving (LNSG) and Rupestris stem pitting (RSP) are a serious problem for the grapevine industry world-wide. The diseases affect yield, rooting ability and graft take of grapevines. Infected grapevines may decline and die within a few years from planting. Eradication of these diseases from vineyards is very difficult as they frequently remain latent and the existing method of detection by biological indexing is time-consuming and expensive.
There is growing evidence that viruses GVA and GVB from Vitivirus genus are involved in inducing symptoms of KSG and CB respectively. Recent studies of these viruses resulted in development of ELISA (based on serology) and PCR (based on nucleotide sequence data) techniques for their identification in grapevines. Although PCR is undoubtedly more sensitive then ELISA, disadvantages of this technique are lack of automation and recently discovered extensive variation of nucleotide sequence of GVB. The latter is especially dangerous as some strains of GVB (and perhaps GVA) may escape undetected by PCR, even if they occur in high concentration in grapevines. Such strains can then be propagated without being noticed.
An important advantage of ELISA for detection of GVA and GVB is that these viruses are serologically related and that strains of these viruses, which differ in reaction to polyclonal antisera, have not been detected yet. The viruses have identical cryptotope(s), which induce(s) a strong immunological response in virus-immunised rabbits Results suggest that GVA and GVB are serologically related to other Vitiviruses. This may enable detection by a single test of other vitiviruses that are increasingly being detected in grapevines. GVA and GVB-specific antibodies will be purified from rabbit antisera by immunoaffinity, using columns with immobilised capsid proteins of viruses. Purified antibodies will be applied in ELISA.
Presentation(s)
1. Goszczynski, D E, Jooste, A E C and Kasdorf, G G F. 2002. Development of ELISA for simultaneous detection of Grapevine virus A and Grapevine virus B. Winetech Grapevine Virus Workshop, 6 May, Simondium.
2. Goszczynski, D E, Jooste, A E C and Kasdorf G, G F. 2003. ELISA for the detection of Grapevine virus A and Grapevine B: Production and some properties of goat antisera to GVA. Winetech Grapevine Virus Workshop, 5 May, Infruitec, Stellenbosch.
Final report
http://www.sawislibrary.co.za/dbtextimages/finalreport90.pdf
Marais, J. 1995. Sauvignon blanc cultivar aroma - a review. South African Journal of Enology + Viticulture. v. 15 n. 2 p. 41-45
Article
Marais, J. 1996. Pienkwordingsvatbaarheid van Suid-Afrikaanse Sauvignon blanc wyne. Wynboer. Mnth Feb p. 3
Marais, J. 1997. Druiftemparatuur: 'n Belangrike kwaliteitsparameter. Wynboer. Mnth Oct p. 8-9
Marais, J. 1997. Voorkoming van pienkwording van Sauvignon blanc-wyn. Wynboer. Mnth Nov p. T11-12
Marais, J. 2001. Effect of grape temperature and yeast strain on Sauvignon blanc wine aroma composition and quality. South African Journal of Enology + Viticulture. v. 22 n. 1 p. 47-51
Swart, E, Marais, J, Britz, T J. 2001. Effect of ascorbic acid and yeast strain on Sauvignon blanc win quality. South African Journal of Enology + Viticulture. v. 22 n. 1 p. 41-46
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