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

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An Erratum to this article was published on 30 January 2017

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.

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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

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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).

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The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. Plant Physiology Laboratory, Department of Botany, Federal University of Santa Catarina, Florianópolis, SC, 88040-900, Brazil

      Neusa Steiner

    2. Graduate Program in Plant Genetic Resources, Department of Plant Science, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, 88040-900, Brazil

      Francine L. Farias-Soares

    3. Plant Cell Biology Laboratory, Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, 88049-900, Brazil

      Éder C. Schmidt, Gladys D. Rogge-Renner & Zenilda L. Bouzon

    4. Laboratory of Plant Developmental Physiology and Genetics, Department of Plant Science, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, 88040-900, Brazil

      Maria L. T. Pereira, Bruna Scheid, Daniela Schmidt & Miguel P. Guerra

    5. Central Laboratory of Electron Microscopy, Federal University of Santa Catarina, Florianópolis, SC, Brazil

      Zenilda L. Bouzon

    6. Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Guiraldes 2160, Pab. 2, Ciudad Universitaria, C1428EGA, Buenos Aires, Argentina

      Sara Maldonado

    7. Department of Biological Sciences, University of Joinville Region, Joinville, SC, Brazil

      Gladys D. Rogge-Renner

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    Correspondence to Neusa Steiner.

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    Handling Editor: Peter Nick

    Neusa Steiner and Francine L. Farias-Soares contributed equally to this work.

    An erratum to this article is available at http://dx.doi.org/10.1007/s00709-017-1080-5.

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    Steiner, N., Farias-Soares, F.L., Schmidt, É.C. et al. Toward establishing a morphological and ultrastructural characterization of proembryogenic masses and early somatic embryos of Araucaria angustifolia (Bert.) O. Kuntze. Protoplasma 253, 487–501 (2016). https://doi.org/10.1007/s00709-015-0827-0

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