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 to 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.