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Morpho-anatomical characterization of mature embryo-derived callus of rice (Oryza sativa L.) suitable for transformation

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Abstract

The objective of this study was to morpho-anatomically characterize embryogenic rice calli during early induction of somatic embryogenesis of three Brazilian rice cultivars. Herein, we explored embryogenic units (EUs) from 2-week-old cut proliferated calli to verify whether they were suitable for Agrobacterium tumefasciens-mediated transformation. Histological analysis and scanning electron microscopy (SEM) were used to analyze these types of calli during early rice callogenesis in the cultivars BRS Primavera, BRS Bonança, and BRS Caiapó. The characteristics of the embryogenic cells were preserved in the EUs, which showed a globular, compact structure that contained tightly packed cells and thus rendered the cells suitable for transformation. The EUs of BRS Caiapó also maintained the characteristics of the non-embryogenic callus, such as an elongated morphology and a lack of cellular organization. In general, the observations of the histological sections corresponded with those of the SEM images. The histological analysis suggested that all cultivars used in these experiments have morphogenic potential. The EUs from proliferated 2-week-old cut calli maintained their embryogenic features. The EUs were subjected to Agrobacterium-mediated transformation, which exhibited a regeneration frequency of 58 % for transformed hygromycin-resistant cell lines. These results show that EUs from proliferated 2-week-old cut calli are suitable for plant transformation.

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Abbreviations

ECM:

Extracellular matrix

ECMSN:

Extracellular matrix surface network

EUs1:

Embryogenic units

EUs2:

Proliferated 2-week-old cut calli

IM:

Induction medium

MS:

Murashige and Skoog

SEM:

Scanning electron microscopy

2,4-D:

2,4-Dichlorophenoxyacetic acid

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Acknowledgments

We thank the Universidade Federal de Goiás for the use of the LabMIC and the Laboratório de Morfologia e Anatomia de Plantas. This work was funded by Embrapa and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Embrapa Arroz e Feijão, Rodovia GO 462, km 12, 75375-000, Santo Antônio de Goiás, Goiás, Brazil

    R. Bevitori

  2. Department of Plant Cytology and Embryology, Jagiellonian University, 9 Gronostajowa St., 30-387, Cracow, Poland

    M. Popielarska-Konieczna

  3. Universidade Federal de Goiás, Goiânia, Goiás, 74001-940, Brazil

    E. M. dos Santos & S. Petrofeza

  4. Embrapa Recursos Genéticos e Biotecnologia, Brasília, DF, 70770-917, Brazil

    M. F. Grossi-de-Sá

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  1. R. Bevitori
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Correspondence to R. Bevitori.

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Bevitori, R., Popielarska-Konieczna, M., dos Santos, E.M. et al. Morpho-anatomical characterization of mature embryo-derived callus of rice (Oryza sativa L.) suitable for transformation. Protoplasma 251, 545–554 (2014). https://doi.org/10.1007/s00709-013-0553-4

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  • DOI: https://doi.org/10.1007/s00709-013-0553-4

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