Grapevine cultivation is penalized by pathological problems with significant impact on production, quality and related costs. This species is affected by numerous viral diseases, such as “fanleaf (GFLV)” and “leaf roll (GLRaV)” diseases, which are the most diffused in Europe. The application of rigorous certification criteria is the only strategy available to control the diffusion of viruses. Traditional breeding techniques are limited in that there is reduced genetic resources and an increase in variability that is unacceptable for the preservation of traditional grapevine clones. Post Transcriptional Gene Silencing (PTGS) has emerged as alternative tool to induce resistance to virus in several plant species, even by using rootstocks as a source of RNAi controlling plant virus infection. For the application of this technology in grapevine it is really important to have efficient regeneration and transformation protocols for the most important cultivars and rootstocks. For this aim, the regeneration and transformation protocol via organogenesis (Mezzetti et al., 2002) was optimized for different grapevine cultivars (Vermentino, Albana, Pignoletto, SanGiovese), in comparison with the efficient table grape cultivar Thompson Seedless, and rootstocks (1103 Paulsen, 110 Richter and Kober 5BB). The meristematic bulks created for each clone were used as explants for Agrobacterium-mediated genetic transformation protocols with a gene construct that express the e-GFP as marker gene. Genotypes having the highest regeneration and transformation efficiency were also used for transformation experiments using a hairpin gene construct designed to silence the RNA-dependent RNA polymerase (RpRd) of the GFLV and GLRaV3, which would induce multiple virus resistances.

IMPLEMENTATION OF A REGENERATION AND TRANSFORMATION PROTOCOL FOR VITIS VINIFERA VARIETIES AND ROOTSTOCK TO INDUCE GENE SILENCING AGAINST GFLV-GLRaV VIRUS

B. MOLESINI;T. PANDOLFINI;
2018-01-01

Abstract

Grapevine cultivation is penalized by pathological problems with significant impact on production, quality and related costs. This species is affected by numerous viral diseases, such as “fanleaf (GFLV)” and “leaf roll (GLRaV)” diseases, which are the most diffused in Europe. The application of rigorous certification criteria is the only strategy available to control the diffusion of viruses. Traditional breeding techniques are limited in that there is reduced genetic resources and an increase in variability that is unacceptable for the preservation of traditional grapevine clones. Post Transcriptional Gene Silencing (PTGS) has emerged as alternative tool to induce resistance to virus in several plant species, even by using rootstocks as a source of RNAi controlling plant virus infection. For the application of this technology in grapevine it is really important to have efficient regeneration and transformation protocols for the most important cultivars and rootstocks. For this aim, the regeneration and transformation protocol via organogenesis (Mezzetti et al., 2002) was optimized for different grapevine cultivars (Vermentino, Albana, Pignoletto, SanGiovese), in comparison with the efficient table grape cultivar Thompson Seedless, and rootstocks (1103 Paulsen, 110 Richter and Kober 5BB). The meristematic bulks created for each clone were used as explants for Agrobacterium-mediated genetic transformation protocols with a gene construct that express the e-GFP as marker gene. Genotypes having the highest regeneration and transformation efficiency were also used for transformation experiments using a hairpin gene construct designed to silence the RNA-dependent RNA polymerase (RpRd) of the GFLV and GLRaV3, which would induce multiple virus resistances.
2018
Vitis vinifera, gene silencing, virus resistance, GFLV-GLRaV
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/991469
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