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An improved protocol for somatic embryogenesis and plant regeneration in macaw palm (Acrocomia aculeata) from mature zygotic embryos

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Abstract

An improved protocol for plant regeneration via somatic embryogenesis was developed using mature macaw palm (Acrocomia aculeata) zygotic embryos as initial explant. For induction of the embryogenic callus (EC), two basic media (BM) were tested consisting of Murashige and Skoog and Eeuwens (Y3) salts with 30 g L−1 sucrose, 0.5 g L−1 glutamine and 2.5 g L−1 Phytagel. The 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6-trichloro-picolinic acid (picloram) and 2,4-dichlorophenoxyacetic acid (2,4-D) auxins were added to the culture media at concentrations of 0, 1.5 or 3.0 mg L−1. After 240 days, the embryogenic calli were transferred to the respective BM media with auxin concentrations reduced to 0.5 or 1.0 mg L−1 in order to differentiate the somatic embryos (SEs). Plant regeneration was performed on the BM media without growth regulators. Embryogenic calli were observed after 180 days of culture and in all treatments with auxin. The Y3 medium showed the best EC formation results (60.8 %). These calli showed yellowish coloration, compact consistency and nodular aspect. After 60 days in differentiation medium, SEs were verified in different stages of development. Histological analysis showed that the SEs were formed from a nodular EC. The SEs generally presented unicellular origin with suspensor formation, and at the end of development, bipolar embryos were observed. The plant regeneration frequency reached levels up to 31.9 % when using induction medium consisting of Y3 associated to 1.5 mg L−1 of 2,4-D and the subsequent auxin reduction to 0.5 mg L−1 in the differentiation stage. Regenerated plants showed normal development, with root and aerial part growth.

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Abbreviations

BM:

Basic media

Dicamba:

3,6-Dichloro-2-methoxybenzoic acid

EC:

Embryogenic callus

MS:

Murashige and Skoog (1962)

Picloram:

4-Amino-3,5,6-trichloro-picolinic acid

SE(s):

Somatic embryo(s)

Y3:

Eeuwens (1976)

ZE:

Zygotic embryos

2,4-D:

2,4-Dichlorophenoxyacetic acid

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Acknowledgments

The authors thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Financiadora de Estudos e Projetos (Finep Grant No. 01.08.0597.01), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes/Embrapa 001-2011/Grant No. 39) for financial support and fellowships.

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Authors and Affiliations

  1. Institute of Biology, PPGBOT, University of Brasília, Brasília, DF, 70910-900, Brazil

    Zanderluce Gomes Luis

  2. Embrapa Genetic Resources and Biotechnology, Av. W5 Norte (final), Brasília, DF, 70770-917, Brazil

    Jonny Everson Scherwinski-Pereira

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  1. Zanderluce Gomes Luis
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Correspondence to Jonny Everson Scherwinski-Pereira.

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Luis, Z.G., Scherwinski-Pereira, J.E. An improved protocol for somatic embryogenesis and plant regeneration in macaw palm (Acrocomia aculeata) from mature zygotic embryos. Plant Cell Tiss Organ Cult 118, 485–496 (2014). https://doi.org/10.1007/s11240-014-0500-x

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