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Protoplasma

, Volume 253, Issue 2, pp 487–501 | Cite as

Toward establishing a morphological and ultrastructural characterization of proembryogenic masses and early somatic embryos of Araucaria angustifolia (Bert.) O. Kuntze

  • Neusa Steiner
  • Francine L. Farias-Soares
  • Éder C. Schmidt
  • Maria L. T. Pereira
  • Bruna Scheid
  • Gladys D. Rogge-Renner
  • Zenilda L. Bouzon
  • Daniela Schmidt
  • Sara Maldonado
  • Miguel P. Guerra
Original Article

Abstract

Somatic embryogenesis is a morphogenetic route useful for the study of embryonic development, as well as the large-scale propagation of endangered species, such as the Brazilian pine (Araucaria angustifolia). In the present study, we investigated the morphological and ultrastructural organization of A. angustifolia somatic embryo development by means of optical and electron microscopy. The proembryogenic stage was characterized by the proliferation of proembryogenic masses (PEMs), which are cellular aggregates composed of embryogenic cells (ECs) attached to suspensor-like cells (SCs). PEMs proliferate through three developmental stages, PEM I, II, and III, by changes in the number of ECs and SCs. PEM III-to-early somatic embryo (SE) transition was characterized by compact clusters of ECs growing out of PEM III, albeit still connected to it by SCs. Early SEs showed a dense globular embryonic mass (EM) and suspensor region (SR) connected by embryonic tube cells (TCs). By comparison, early somatic and zygotic embryos showed similar morphology. ECs are round with a large nucleus, nucleoli, and many cytoplasmic organelles. In contrast, TCs and SCs are elongated and vacuolated with cellular dismantling which is associated with programmed cell death of SCs. Abundant starch grains were observed in the TCs and SCs, while proteins were more abundant in the ECs. Based on the results of this study, a fate map of SE development in A. angustifolia is, for the first time, proposed. Additionally, this study shows the cell biology of SE development of this primitive gymnosperm which may be useful in evolutionary studies in this area.

Keywords

Somatic embryogenesis Histochemistry Pluripotency Suspensor cell Conifers 

Abbreviations

ABA

Abscisic acid

CA

Cytochemical analysis

CBB

Coomassie Brilliant Blue

CLSM

Confocal laser scanning microscopy

DAPI

4′,6-Diamidino-2-phenylindole dihydrochloride

ECs

Embryogenic cells

EM

Embryonic mass

FLD

Fluridone

LM

Light microscopy

PAS

Periodic acid-Schiff

PCD

Programmed cell death

PEG

Polyethylene glycol 3350

PEM

Proembryogenic mass

PEMs

Proembryogenic masses

PGRs

Plant growth regulators

SCs

Suspensor-like cells

SR

Suspensor region

SE

Somatic embryo

SEM

Scanning electron microscopy

TB-O

Toluidine blue

TCs

Embryonic tube cells

TEM

Transmission electron microscopy

Notes

Acknowledgments

The authors acknowledge the staff of the Central Laboratory of Electron Microscopy (LCME) of the Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil. This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil) and the Fundação de Apoio à Pesquisa Cientifica e Inovação Tecnológica do Estado de Santa Catarina (FAPESC).

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Neusa Steiner
    • 1
  • Francine L. Farias-Soares
    • 2
  • Éder C. Schmidt
    • 3
  • Maria L. T. Pereira
    • 4
  • Bruna Scheid
    • 4
  • Gladys D. Rogge-Renner
    • 3
    • 7
  • Zenilda L. Bouzon
    • 3
    • 5
  • Daniela Schmidt
    • 4
  • Sara Maldonado
    • 6
  • Miguel P. Guerra
    • 4
  1. 1.Plant Physiology Laboratory, Department of BotanyFederal University of Santa CatarinaFlorianópolisBrazil
  2. 2.Graduate Program in Plant Genetic Resources, Department of Plant ScienceFederal University of Santa CatarinaFlorianópolisBrazil
  3. 3.Plant Cell Biology Laboratory, Department of Cell Biology, Embryology and GeneticsFederal University of Santa CatarinaFlorianópolisBrazil
  4. 4.Laboratory of Plant Developmental Physiology and Genetics, Department of Plant ScienceFederal University of Santa CatarinaFlorianópolisBrazil
  5. 5.Central Laboratory of Electron MicroscopyFederal University of Santa CatarinaFlorianópolisBrazil
  6. 6.Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y NaturalesUniversidad de Buenos AiresBuenos AiresArgentina
  7. 7.Department of Biological SciencesUniversity of Joinville RegionJoinvilleBrazil

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