Two-dimensional electrophoretic analysis of proteins associated with somatic embryogenesis development in Cupressus sempervirens L.

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 polyembryogenes is phenomenon which maintained them in a continuous proliferating status in stead of undergoing a complete embryogenic development. Only the addition o f bovine serum albumin to the culture allowed somatic embryo development an d maturation. Major histological differences were noticed between developin g and nondeveloping embryogenic cultures. Attempts to find proteins that co uld be associated with developmental stages of somatic embryos have been ac hieved. Proteins were extracted and analyzed by two-dimensional electrophor esis from nondeveloping embryogenic cultures (SO) and from embryogenic cult ures at three different stages of somatic embryo development: small size an d rounded shape embryos (S1), increased size embryos with a well-developed suspensor (S2) and embryos with two well-separated cotyledons (S3). The res ults 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 inv olved in somatic embryo development and maturation. Specific polypeptides a ssociated with the presence of bovine serum albumin (BSA) in the medium hav e been detected.