ISHS Acta Horticulturae 692: II International Symposium on Biotechnology of Tropical and Subtropical Species
GERMPLASM CONSERVATION, VIRUS ERADICATION AND SAFE STORAGE OF TRANSFORMATION COMPETENT CULTURES IN BANANA: THE IMPORTANCE OF CRYOPRESERVATION |
| Authors: | B. Panis, B. Helliot, H. Strosse, S. Remy, P. Lepoivre, R. Swennen |
| Keywords: | Musa spp., germplasm collections, meristem culture, embryogenic cell suspensions |
| Abstract:
Currently cryopreservation is applied mainly for safe long-term germplasm conservation of seedless and thus vegetatively propagated crops like banana (Musa spp.). Three cryopreservation methods for shoot-tip cultures of banana are currently available. The first method relies on rapid freezing of highly proliferating meristem cultures precultured for 2 weeks on 0.4 M sucrose. The second method is based on vitrification of tiny meristems excised from rooted in vitro plants. The third, and until now most successful protocol, is a combination of the previous ones; vitrification of highly proliferating, sucrose-precultured meristem cultures. Post-thaw regeneration rates are up to 75 %, depending on the cryopreservation protocol and the cultivar. Besides its traditional application for germplasm storage, cryopreservation of meristem cultures can also result in virus eradication of BSV and CMV from infected plants. Only the most meristematic, and thus the least virus-infected, part of these cultures regenerates after freezing. Finally, cryopreservation can also be applied to plant material with specific characteristics, such as medicinal and alcohol producing cell lines, genetically transformed tissues and transformation competent tissues. The safe storage of transformation competent embryogenic cell suspensions of banana is of utmost importance since their initiation is difficult and time-consuming and their morphogenic capacity decreases with time. Cell cultures were recovered after 4 years of storage in liquid nitrogen. We showed that viability and regeneration capacity remainded intact, as well as competence to Agrobacterium mediated transformation. | |
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