Utilization of Single Nodal Cuttings and Therapies for Eradicating Double-Infected Potato Virus (PLRV, PVY) from In Vitro Plantlets of Potato (Solatium tuberosutri)

摘要

In vitro plantlets of potato genotype 'F9-99' infected with PLRV and PVY were used for the virus eradication using heat and chemical treatments. Subculture medium was added with acetylsalicylic acid (ASA) (10~(-6), 10~(-5), and 10~(-4)M) and ribavirin(10, 15, and 20 mg.L~(-1)) alone or in combinations. The plantlets were incubated at 25°C (control), and alternating heat treatments 35/30°C for 1, 4, and 8 h or 42/25°C for 4 h. Virus status of the plantlets were examined by means of double antibodysandwich-enzyme linked immunosorbent assay and reverse transcription polymerase chain reaction. After 4-8 weeks of treatment, 40, 51, 41, 36, and 0% survival were observed in the control (MS and 25°C), ASA plus ribavirin (25°C), MS plus heat treatment(35/30°C), ASA plus ribavirin (35/30°C), and MS medium with heat treatment (42/25°C), respectively. As for virus eradication, heat treatment (35/30°C) with 1 h alternating thermo-cycle resulted in 100% PVY elimination, but it was much more difficultto remove the PLRV from the double-infected potato plantlets with PVY and PLRV. The highest rate of PLRV eradication was 60% from the combined treatment of 10~(-5) M ASA, 20 mg-L~(-1) ribavirin, and heat treatment at 35/30°C for 8 h alternating temperature.