biotechnology - SimpBTH - Universitatea De Stiinte Agronomice Si ...
biotechnology - SimpBTH - Universitatea De Stiinte Agronomice Si ...
biotechnology - SimpBTH - Universitatea De Stiinte Agronomice Si ...
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4 th International Symposium of Biotechnology10 – 11 th November 2011, Bucharest, RomaniaP.I.8. EFFECT OF DIRECT ALTERNATING CURRENTSTIMULATION ON VIRUS – ELIMINATION IN GRAPEVINEIonela Cătălina GUŢĂ 1 , Elena-Cocuţa BUCIUMEANU 1 , Benedict OPRESCU 2 , LaviniaTĂTARU 21 National Research& <strong>De</strong>velopment Institute for Biotechnology in Horticulture, 37 Bucureşti-PiteştiRoad, 117715-Ştefăneşti-Argeş, România, Tel./Fax.: 0248-866214,E-mail: gutaionelacatalina@yahoo.com; ebuciumeanu@yahoo.com2 University of Piteşti, Faculty of Science, 1 Târgu din Vale, 110040-Piteşti, România,Tel.: 0348-453100, Fax: 0348-453123, E-mail: b.oprescu@yahoo.com; lavdiana@yahoo.comKeywords: Vitis, virus, eradication, electrotherapyThe meristem type culture and thermotherapy are individually or in combination the mostfrequent methods of obtaining virus-free plants. The application of heat treatment for viruseliminationin plants is time and energy consuming and needs special equipments (chambers for heattreatment). In the case of the meristem culture, the percent of virus elimination is influenced by thegenotype, the type of the virus and especially of the difficulty of meristem excision (the percent ofvirus elimination is inverse proportion to the explants dimensions).The use of electrotherapy with discontinuous electric current followed by in vitro culturewas investigated as alternative technique to eliminate grapevine viruses, in simple or mixed infectionsat a time, in seven Vitis vinifera L. varieties naturally infected. The study had in view the mostdangerous viruses of this crop: fanleaf virus (GFLV), arabis mosaic virus (ArMV), leafroll associatedvirus 1 (GLRaV-1), leafroll associated virus 3 (GLRaV-3), and fleck virus (GFkV). Alternatingelectric current of 1; 100; 1000 and 10000 kHz was applied for 5; 10 and 20 min at the cut ends ofone budded herbaceous cuttings. Treated axillary buds were grown on MS-medium containinggrowth regulators. The efficacy of virus elimination process was assessed by enzyme-linkedimmunosorbent assay (ELISA) testing of regenerated acclimated grapevine plants. The analysisshowed some encouraging results for elongated viruses GLRaV-1 and GLRaV-3 in single infectionsand no virus eradication in mixed infections had been obtained. The electrotherapy of GLRaV-1-infected material produced maximum 12,5% virus-free plants at 1000 kHz for 10 min of exposure,and 4,5% GLRaV-3-free vines at 10000 kHz for 10 min of exposure. The result regarding GLRaV-1elimination was not reproducible in the case of mixed infection containing it. No satisfactory resultshave been achieved for isometric viruses eradication (GFLV, ArMV, GFkV) either in simple or mixedinfections. This treatement did not influenced significantly the in vitro regeneration process ofgrapevine.38