[A Comparative Study Between Different Methods To Produce In vitro Virus Y-free Potato seeds]
2011
Massa,R.
This study was conducted to obtain a virus-free propagation material from two cultivars of potato (Burren and Binella) infected with potato virus Y (PVY), by using the meristem culture technique with three different therapies, thermo-, chemo- and electrotherapies. The Meristem (lengths were 0.1, 0.2, 0.3 μm) were excised from infected plants, solidified MS medium containing 30 g/l sucrose, and supplemented with Vitamins (5 mg/l Ascorbic acid, 5 mg/l Pyridoxine, 5 mg/l Nicotinic acid, 5 mg/l Thiamine, 200 mg/l Inositole) and Hormones ( 2 mg/l GA3 , 0.2 mg/l kinetine) was used. Virus status of in vitro plantlets was determined by double antibody sandwich enzyme linked immunosorbent assay (DASELISA). Results showed that the usage of the size 0.1μm gives the highest rate of virus-free plants. After treatment of thermotherapy at approximately 37±2°C during 40 days, the highest rate of PVY elimination achieved when the meristem in length of (100 μ m) were excised (75%) in Binella and (69%) in Burren. These results indicated that thermotherapy is moderately efficient for PVY elimination. Chemotherapy was undertaken with (10-20-30 ) mg L-1 ribavirin (RBV) , The highest numbers of virus free plantlets 87% in Binella and 82% in Burren were obtained from the ribavirin's concentration (20 mg/ l) combined with meristem-tips in length (100μm). This study indicated that PVY was not completely eliminated by the treatment of ribavirin alone and showed severe growth abnormalities specially when we added the ribavirin (30 mg/l) to the meristem's medium . Finally, the highest rates of PVY (93%in Binella and 87% in Burren) free plantlets were obtained from meristem-tips in length 100μ m excised after electric treatments (15 mA /10 min). so our study indicated that electric shock had higher efficiency for PVY elimination.
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