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The relationship between cell expansion and cell cycling during somatic embryogenesis was studied in cultured bent-cotyledon-stage zygotic embryos of a transgenic stock of Arabidopsis thaliana harboring a cyclin 1 At:β-glucuronidase (GUS) reporter gene construct. In embryos cultured in a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), following a brief period of growth by cell expansion, divisions were initiated in the procambial cells facing the adaxial side at the base of the cotyledons. Cell division activity later spread to almost the entire length of the cotyledons to form a callus on which globular and heart-shaped embryos appeared in about 10 d after culture. Anatomical and morphogenetic changes observed in cultured embryos were correlated with patterns of cell cycling by histochemical detection of GUS-expressing cells. Although early-stage somatic embryos did not develop further during their continued growth in the auxin-containing medium, maturation of embryos ensued upon their transfer to an auxin-free medium. In a small number of cultured zygotic embryos the shoot apical meristem was found to differentiate a leaf, a green tubular structure, or a somatic embryo. Contrary to the results from previous investigations, which have assigned a major role for the shoot apical meristem and cells in the axils of cotyledons in the development of somatic embryos on cultured zygotic embryos of A. thaliana, the present work shows that somatic embryos originate almost exclusively on the callus formed on the cotyledons. Other observations such as the induction of somatic embryos on cultured cotyledons and the inability of the embryo axis (consisting of the root, hypocotyl, and shoot apical meristem without the cotyledons) to form somatic embryos, reaffirm the important role of the cotyledons in somatic embryogenesis in this plant.

Contents Summary  000 Introduction  000 Discovery of double fertilization  000 Seed development without double fertilization  000 A case for double fertilization in gymnosperms  000 Structural and cytological perspectives on double fertilization  000 In vitro double fertilization  000 Genetic and molecular perspectives  000 Concluding comments  000 Acknowledgements  000 References  000 Summary The fusion of one sperm with the egg cell to form the embryo and of the other sperm with the polar fusion nucleus to give rise to the endosperm (‘double fertilization’) was discovered by Nawaschin in 1898 in the liliaceous plants, Lilium martagon and Fritillaria tenella. The occurrence of two fusion events analogous to double fertilization has recently been described in some gymnosperm species although the product of the second fusion is a transient embryo, rather than the endosperm as in angiosperms. Recent investigations in angiosperms describe the cell biology and nuclear cytology of double fertilization and the successful in vitro demonstration of the two fusion events using isolated egg cells, central cells, and sperm cells and the development of the fusion products into the embryo and endosperm. Molecular and genetic studies on the component elements of double fertilization have focused on the identification of mutants of Arabidopsis thaliana that display developmental patterns in the seed that result in autonomous endosperm development and even partial embryogenesis in the absence of fertilization. Characterization of the genes and their protein products has provided evidence for a predominant effect of maternal gametophytic genes and of silencing of paternal genes during double fertilization.

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Objective Reinventing data extraction from electronic health records (EHRs) to meet new analytical needs is slow and expensive. However, each new data research network that wishes to support its own analytics tends to develop its own data model. Joining these different networks without new data extraction, transform, and load (ETL) processes can reduce the time and expense needed to participate. The Informatics for Integrating Biology and the Bedside (i2b2) project supports data network interoperability through an ontology-driven approach. We use i2b2 as a hub, to rapidly reconfigure data to meet new analytical requirements without new ETL programming.Materials and Methods Our 12-site National Patient-Centered Clinical Research Network (PCORnet) Clinical Data Research Network (CDRN) uses i2b2 to query data. We developed a process to generate a PCORnet Common Data Model (CDM) physical database directly from existing i2b2 systems, thereby supporting PCORnet analytic queries without new ETL programming. This involved: a formalized process for representing i2b2 information models (the specification of data types and formats); an information model that represents CDM Version 1.0; and a program that generates CDM tables, driven by this information model. This approach is generalizable to any logical information model.Results Eight PCORnet CDRN sites have implemented this approach and generated a CDM database without a new ETL process from the EHR. This enables federated querying within the CDRN and compatibility with the national PCORnet Distributed Research Network.Discussion We have established a way to adapt i2b2 to new information models without requiring changes to the underlying data. Eight Scalable Collaborative Infrastructure for a Learning Health System sites vetted this methodology, resulting in a network that, at present, supports research on 10 million patients’ data.Conclusion New analytical requirements can be quickly and cost-effectively supported by i2b2 without creating new data extraction processes from the EHR.