Background: A protocol for micropropagation of the grape (Vitis vinifera L.) cultivar ‘Monastrell' was developed. Initial plant material was based on sanitary selection of grapevine plants performed by real-time RT-PCR, to confirm the absence of Grapevine fanleaf virus (GFLV), Arabis mosaic virus (ArMV), Grapevine leafroll-associated virus 1 (GLRaV-1), Grapevine leafroll-associated virus 3 (GLRaV-3), and Grapevine fleck virus (GFkV).

Results: The effects of the salt composition (comparing Lloyd and McCown woody plant medium and Murashige and Skoog medium ½ macronutrients) and the growth regulator benzylaminopurine (BAP), at 0 and 8.9 µM, on the multiplication step were evaluated using nodes as explants. The most-efficient procedure consisted of bud induction in the medium with Lloyd and McCown woody plant salts and 8.9 µM BAP for 30 days along with elongation in cytokinin-free medium for 60 days, which gave 22 nodes/explant (174 plants/initial plant). A second cycle of multiplication in a medium without the growth regulator for another 60 days could give around 10,000 nodes; this quantity of rooted plants could be obtained after another two months of culture. All plants acclimatized after the second cycle of multiplication were successfully transferred to soil.

Conclusion: We developed an optimal protocol for V. vinifera cv. Monastrell micropropagation, the first described for this cultivar.