title: Label-Free Quantitative Proteomics of Embryogenic and Non-Embryogenic Callus during Sugarcane Somatic Embryogenesis.
abstract: The development of somatic cells in to embryogenic cells occurs in several stages and ends in somatic embryo formation, though most of these biochemical and molecular changes have yet to be elucidated. Somatic embryogenesis coupled with genetic transformation could be a biotechnological tool to improve potential crop yields potential in sugarcane cultivars. The objective of this study was to observe somatic embryo development and to identify differentially expressed proteins in embryogenic (E) and non-embryogenic (NE) callus during maturation treatment. E and NE callus were cultured on maturation culture medium supplemented with different concentrations (0.0, 0.75, 1.5 and 2.0 g L(-1)) of activated charcoal (AC). Somatic embryo formation and differential protein expression were evaluated at days 0 and 21 using shotgun proteomic analyses. Treatment with 1.5 g L(-1) AC resulted in higher somatic embryo maturation rates (158 somatic embryos in 14 days) in E callus but has no effect in NE callus. A total of 752 co-expressed proteins were identified through the SUCEST (The Sugarcane EST Project), including many housekeeping proteins. E callus showed 65 exclusive proteins on day 0, including dehydrogenase, desiccation-related protein, callose synthase 1 and nitric oxide synthase. After 21 days on maturation treatment, 14 exclusive proteins were identified in E callus, including catalase and secreted protein. NE callus showed 23 exclusive proteins on day 0 and 10 exclusive proteins after 21 days on maturation treatment, including many proteins related to protein degradation. The induction of maturation leads to somatic embryo development, which likely depends on the expression of specific proteins throughout the process, as seen in E callus under maturation treatment. On the other hand, some exclusive proteins can also specifically prevent of somatic embryos development, as seen in the NE callus.
authors.affiliation: Laboratório de Biotecnologia, Centro de Biociências e Biotecnologia (CBB), Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF). Campos dos Goytacazes, RJ, Brazil.
authors.forename: Angelo Schuabb
authors.initials: AS
authors.lastname: Heringer
authors.affiliation: Laboratório de Biotecnologia, Centro de Biociências e Biotecnologia (CBB), Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF). Campos dos Goytacazes, RJ, Brazil.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Barroso
authors.affiliation: Laboratório de Biologia Celular de Plantas, Instituto de Biociências, Universidade de São Paulo (USP), São Paulo, SP, Brazil.
authors.forename: Amanda Ferreira
authors.initials: AF
authors.lastname: Macedo
authors.affiliation: Laboratório de Biologia Celular e Tecidual, CBB-UENF, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Claudete
authors.initials: C
authors.lastname: Santa-Catarina
authors.affiliation: Mass Spectrometry Research and Development Laboratory, Waters Corporation, São Paulo, SP, Brazil.
authors.forename: Gustavo Henrique Martins Ferreira
authors.initials: GH
authors.lastname: Souza
authors.affiliation: Laboratório de Biologia Celular de Plantas, Instituto de Biociências, Universidade de São Paulo (USP), São Paulo, SP, Brazil.
authors.forename: Eny Iochevet Segal
authors.initials: EI
authors.lastname: Floh
authors.affiliation: Laboratório de Biotecnologia, Centro de Biociências e Biotecnologia (CBB), Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF). Campos dos Goytacazes, RJ, Brazil.
authors.forename: Gonçalo Apolinário
authors.initials: GA
authors.lastname: de Souza-Filho
authors.affiliation: Laboratório de Biotecnologia, Centro de Biociências e Biotecnologia (CBB), Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF). Campos dos Goytacazes, RJ, Brazil.
authors.forename: Vanildo
authors.initials: V
authors.lastname: Silveira
mesh_terms: D010940:Plant Proteins / Q000378:metabolism*; D058439:Plant Somatic Embryogenesis Techniques / Q000379:methods*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D031786:Saccharum / Q000196:embryology* / Q000378:metabolism*; D012639:Seeds / Q000254:growth & development / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D010940:Plant Proteins; D020543:Proteome