EXTRA SPINDLE POLES (Separase) controls anisotropic cell expansion in Norway spruce (Picea abies) embryos independently of its role in anaphase progression

The caspase-related protease separase (EXTRA SPINDLE POLES, ESP) plays a major role in chromatid disjunction and cell expansion in Arabidopsis thaliana. Whether the expansion phenotypes are linked to defects in cell division in Arabidopsis ESP mutants remains elusive. Here we present the identification, cloning and characterization of the gymnosperm Norway spruce (Picea abies, Pa) ESP. We used the P. abies somatic embryo system and a combination of reverse genetics and microscopy to explore the roles of Pa ESP during embryogenesis. Pa ESP was expressed in the proliferating embryonal mass, while it was absent in the suspensor cells. Pa ESP associated with kinetochore microtubules in metaphase and then with anaphase spindle midzone. During cytokinesis, it localized on the phragmoplast microtubules and on the cell plate. Pa ESP deficiency perturbed anisotropic expansion and reduced mitotic divisions in cotyledonary embryos. Furthermore, whilst Pa ESP can rescue the chromatid nondisjunction phenotype of Arabidopsis ESP mutants, it cannot rescue anisotropic cell expansion. Our data demonstrate that the roles of ESP in daughter chromatid separation and cell expansion are conserved between gymnosperms and angiosperms. However, the mechanisms of ESP-mediated regulation of cell expansion seem to be lineage-specific

Involvement of Plant Hormones and Plant Growth Regulators on in vitro Somatic Embryogenesis

In spite of the importance attained by somatic embryogenesis and of the many studies that have been conducted on this developmental process, there are still many aspects that are not fully understood. Among those features, the involvement of plant hormones and plant growth regulators on deTermining the conversion of somatic onto embryogenic tissues, and on allowing progression and maturation of somatic embryos, are far away from being completely comprehended. Part of these difficulties relies on the frequent appearance of contradictory results when studying the effect of a particular stimulus over a specific stage in somatic embryogenesis. Recent progress achieved on understanding the interaction between exogenously added plant growth regulators over the concentration of endogenous hormones, together with the involvement of sensitivity of the tissues to particular hormone groups, might help clarifying the occurrence of divergent patterns in somatic embryogenesis, and in tissue culture in general. The aspects described above, emphasizing on the effect of the concentration of plant hormones and of the addition of plant growth regulators during the different phases of somatic embryogenesis, will be reviewed in this paper. Citations will be limited to review articles as much as possible and to individual articles only in those cases in which very specific or recent information is presented.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro para Investigaciones en Granos y Semillas (CIGRAS