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Annals of Botany 74: 547-555, 1994
© 1994 Annals of Botany Company

Cryopreservation of Somatic Embryos: A Tool for Germplasm Storage and Commercial Delivery of Selected Plants

H. Tessereau, B. Florin, M. C. Meschine, C. Thierry and V. Pétiard

Francereco S.A., 101 Avenue Gustave Eiffel, 37390 Notre Dame d'Oé, France

The development of long-term preservation methods of somatic embryos was investigated as a contribution to synthetic seed technology, Thus, cryopreservation in liquid nitrogen was attempted with somatic embryos of two species. Coffea canephora Pierre and Daucus carota L. Cryopreservation of carrot embryos simply required a 21-h sucrose pretreatment followed by a prefreezing step at -20°C. A freeze-hardening treatment consisting of embryo culture in the presence of high sucrose molarity and 1 µM ABA was developed for coffee embryos. Following desiccation at 75% relative humidity (RH) and +24°C for 7 d, embryos were frozen by rapid immersion in liquid nitrogen. Direct regrowth of frozen-thawed embryos was obtained for the majority of both carrot and coffee embryos. Even large carrot embryos were able to survive cryopreservation. The size of embryos and the age of the embryogenic strain from which the embryos originated determined their ability to develop into normal plantlets. Direct regrowth without any secondary callogenesis and/or embryogenesis is a prerequisite for practical use of preserved embryos either in a nursery or through direct planting.Copyright 1994, 1999 Academic Press

Coffea canephora Pierre, Daucus carota L. cryopreservation, desiccation, sucrose, abscisic acid, somatic embryos, synthetic seeds


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