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Two-dimensional gel electrophoretic analysis and histological studies were performed on somatic embryos in cypress. Embryogenic cultures were obtained from in vitro culture of immature seeds. On a modified Murashige and Skoog (MS) medium they showed an intense and repetitive cleavage polyembryogenesis phenomenon which maintained them in a continuous proliferating status instead of undergoing a complete embryogenic development. Only the addition of bovine serum albumin to the culture allowed somatic embryo development and maturation. Major histological differences were noticed between developing and nondeveloping embryogenic cultures. Attempts to find proteins that could be associated with developmental stages of somatic embryos have been achieved. Proteins were extracted and analyzed by two-dimensional electrophoresis from nondeveloping embryogenic cultures (S0) and from embryogenic cultures at three different stages of somatic embryo development: small size and rounded shape embryos (S1), increased size embryos with a well-developed suspensor (S2) and embryos with two well-separated cotyledons (S3). The results revealed some qualitative and quantitative protein variations between the two cultures. Some could be connected with the induction of pro-embryo differentiation whereas others should be more related to the mechanisms involved in somatic embryo development and maturation. Specific polypeptides associated with the presence of bovine serum albumin (BSA) in the medium have been detected.

The proliferation of parenchymatous foliar tissue of Quercus ilex was obtained from current year leaves taken from old trees and cultivated in vitro on modified Murashige and Skoog's medium supplemented with benzylaminopurine (4 mg ∙ L−1) and naphthyl acetic acid (0.5 mg ∙ L−1). Only the fragments cultured in October reacted. The neoformations only appeared on calluses that had not been subcultured for 7 months. Primary nodules arising on these calluses were removed and subcultured on the same medium either in the dark or in the light. In the dark only, they produced secondary nodules, which were the source of somatic embryos both in light and dark. Presently, they seem to regulate their structure in the dark but they do not develop in a way that leads to germination.

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