A cryopreservation protocol for immature zygotic embryos of species of Ilex (Aquifoliaceae)

MROGINSKI, L.; SANSBERRO, P.; SCOCCHI, A.; LUNA, C.; REY, H

BIOCELL

Facultad de Ciencias Médicas, Universidad Nacional de Cuyo.

Lugar: Mendoza; Año: 2008 vol. 32 p. 33 - 39

0327-9545

    This report presents preliminary results from a study aimed to develop a protocol for fruit cryopreservation and the recovering of plants through zygotic embryo culture, as a tool for  the conservation of Ilex spp. (Aquifoliaceae) genetic resources. Fruits from eight plant species were harvested and maintained at 4ºC for 1 month.  Direct immersion of fruits in nitrogen liquid was lethal to the embryos of all the plant species tested. However, the embryos of five species (Ilex brasiliensis, I. dumosa, I. microdonta, I. paraguariensis, and I. pseudoboxus) dissected from fruits (with a cryoprotective solution containing 50% sucrose and 50% glycerol) cryopreserved using slow freezing method (1ºC.min-1 until – 40ºC) and stored in liquid nitrogen, when cultured in vitro, germinated with frequencies ranging from 3 to 23%. The embryos were cultured in vitro in darkness using a culture medium composed of ¼-strength Murashige and Skoog medium, with 3% sucrose, 0.65% agar and 0.1 mg.L-1 zeatin.     The germinated embryos developed into normal plantlets after 60 days in culture. These results suggest that this methodology might be used for long-term germplasm storage of Ilex species