Cryopreservation for the elimination of cucumber mosaic and banana streak viruses from banana (Musa spp.)

The utilisation of cryopreservation for the eradication of cucumber mosaic virus (CMV) or banana streak virus (BSV) from Musa spp. was investigated. Banana plants, cv. Williams (AAA, Cavendish subgroup), were mechanically infected with CMV or naturally infected with BSV, and proliferating meristems were produced from the infected plants. Excised meristematic clumps were cryopreserved through vitrification using PVS-2 solution. The health status of regenerated in vitro plants was first checked by means of ELISA. The putative virus-free material was subsequently tested a second time following greenhouse acclimatisation. The frequency of virus eradication for CMV and BSV was 30% and 90%, respectively, following cryopreservation. In comparison, the frequency of virus-free plants regenerated directly from highly proliferating meristems, corresponding to a spontaneous eradication rate, reached 0% and 52% for CMV and BSV, respectively. The conventional meristem culture resulted in 0% CMV-free plants and 76% BSV-free plants, while the cryoprotective treatment resulted in 2% CMV-free plants and 87% BSV-free plants. To understand the mode of action of cryopreservation for the eradication of viral particles, we examined the structure of the meristem tips by light microscopy. The cryopreservation method used only allowed survival of small areas of cells located in the meristematic dome and at the base of the primordia.