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Guta I.C.,National Research and Development Institute for Biotechnology in Horticulture | Buciumeanu E.-C.,National Research and Development Institute for Biotechnology in Horticulture
Romanian Biotechnological Letters | Year: 2011

Shoot apices and first axillary buds double infected by leafroll associated virus serotype 1 and vitivirus A, collected from grapevine plants variety Servant, were subjected to in vitro chemotherapy technique. Ribavirin of 80 μmol L-1 added in solid medium was used for 30-90 days followed by 1-3 subcultures on free-drug proliferating medium. The phytotoxic effect evaluated by micropropagation rate, microshoots differentiation and elongation process was observed in the exposure period, but its intensity decreased along with transfer to medium without viricide. Virus-free plants regeneration was assessed repeatedly by enzyme-linked immunosorbent assay, considering the type of ribavirin treated explant from which the new grapevine derived. The plants developed from shoot apices were found 33% negative for GVA but no GLRaV-1- free plant was identified. The plants originated from axillary buds were 100% GVA-free and 11% were GLRaV-1-free. In this case, ribavirin was not appropriate to eliminate the virus complex; low obtained percentage of healthy plants is due to synergistic activity of two viruses and, perhaps, to plant adaptation to multiple viral infection. Healthy plants were further investigated on the beneficial effect of viruses elimination, in terms of dynamics of shoots growth and wood maturation. © 2011 University of Bucharest.


Guta I.C.,National Research and Development Institute for Biotechnology in Horticulture | Buciumeanu E.-C.,National Research and Development Institute for Biotechnology in Horticulture | Visoiu E.,National Research and Development Institute for Biotechnology in Horticulture
Notulae Botanicae Horti Agrobotanici Cluj-Napoca | Year: 2014

Grapevine fleck virus produces a ubiquitous disease, latent in European grapevine varieties (Vitis vinifera L.) and in most American rootstocks, being found in all viticultural countries, in simple or complex infections with other more dangerous viruses. Different techniques for sanitation showed controversial results regarding fleck elimination in grapevine. In vitro culture of 'Tamâioasa româneasca' 3-2-2 with fleck virus and 'Burgund' 63 Mn with double infection (Grapevine fleck virus and Grapevine virus A), naturally infected have been submitted to chemotherapy using a combination of ribavirin and oseltamivir in three concentration variants (V1-40 mg/L ribavirin + 40 mg/L oseltamivir; V2-20 mg/L ribavirin + 40 mg/L oseltamivir; V3-20 mg/L ribavirin + 80 mg/L oseltamivir) and three consecutive subcultures. The plants regenerated after each subculture were evaluated by ELISA from the viewpoint of virus elimination and the RT-PCR was used for confirmation of the diagnostic. Due the phytotoxic effect of viricides, the multiplication rate decreased on experimental variants in the next subculture comparatively to the control, but no mortality of explants has been registered. Grapevine fleck virus has been 100% eliminated both from simple and mixed infections on all variants by in vitro chemotherapy, under the simultaneous action of two viricides. Unsatisfactory results have been achieved with Grapevine virus A elimination.


Gheorghe R.N.,National Research and Development Institute for Biotechnology in Horticulture | Popescu C.F.,National Research and Development Institute for Biotechnology in Horticulture | Pamfil D.,University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca | Ciocirlan C.N.,National Research and Development Institute for Biotechnology in Horticulture | Sestras R.,University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca
Romanian Biotechnological Letters | Year: 2010

The PCR-based DNA microsatellite analysis has been applied to define genetic relationships among eleven Romanian grapevine varieties. 87 alleles with a mean number of 7.90 alleles per locus were detected in the set of cultivars analyzed. The total number of alleles per locus varied between 4 (in VVMD24 and VVIQ52) and 10 (in VMC8G9, UDV125 and VMC5G6.1). The expected heterozygosity (gene diversity) ranged from 0.63 at locus VVMD24 to 0.92 at locus VVMD28. The observed heterozygosity varied between 0.63 at locus VVMD24 to 1 at locus VVMD28 and was higher than the expected one at 4 out of 11 loci. Microsatellite analysis proved to be useful for the genetic characterization of Romanian grapevines, allowing precise identification and good discrimination of all tested cultivars. © 2010 University of Bucharest.

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