Abstract
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 4°C for 1 month. Direct immersion of fruits in liquid nitrogen was lethal to the embryos of all the plant species tested. However, the embryos of five species (Ilex brasiliensis, L dumosa, I. microdonta, I. paraguariensis and I. pseudoboxus) dissected from fruits (with a cryoprotective solution containing 50% sucrose and 50% glycerol), cryopreserved using a slow freezing method (1°C.min -1 until -40°C) 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 1/4-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.
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