Simultaneous elimination of Cucumber mosaic virus and Cymbidium mosaic virus infecting Vanilla planifolia through meristem culture

Abstract

Vanilla (Vanilla planifolia G.jackson), a perennial fleshy vine infected by Cucumber mosaic virus (CMV) and Cymbidium mosaic virus (CymMV) was freed from infection through meristem culture. Apical meristem measuring 0.1-0.25 mm were isolated and cultured in Murashige and Skoog (MS) medium supplemented with 0.45 µM thidiazuron for 40-45 days to initiate the growth. Following enlargement of meristem, it was transferred to MS medium supplemented with 4.43 µM 6-benzyl aminopurine (BAP) and 2.68 µM α-naphthalene acetic acid (NAA) for regeneration. The regenerated plantlets were hardened in insect-free glasshouse and confirmed for the elimination of viruses using reverse transcription PCR (RT-PCR) with virus specific primer pairs. The frequency of CMV elimination was 79.4% while that of CymMV was 82.4% when tested individually. We obtained a frequency of 75% simultaneous elimination of both viruses. This is the first report of meristem culture and virus elimination in vanilla.