Anatomical and histochemical studies of the somatic embryogenesis of <i>Syagrus oleracea</i> from immature inflorescences

Anatomical and histochemical studies were carried out during the somatic embryogenesis of Syagrus oleracea (Mart.) Becc. from immature inflorescences. Immature rachillas were inoculated on Murashige and Skoog medium with 4.52 μM 4-amino-3,5,6-trichloropicolinic acid (Picloram) and 2,4-diclorophenoxyacetic acid. Auxin concentrations were gradually reduced for the formation of somatic embryos, while for plantlet regeneration, embryos were inoculated on a medium without growth regulator. After 270 days, two types of embryogenic calluses were observed in explants under the effect of Picloram: nodular, with a smooth surface and those without definite shape, with an irregular surface, both presenting a meristematic zone with polysaccharide content. Callus progression to the differentiation stage allowed for the conversion of somatic embryos, some with a well-defined protoderm and procambial bundles. The histochemical analysis of the somatic embryos did not identify the presence of polysaccharide and protein reserves, which emphasizes the necessity for optimization of the maturation conditions.

Keywords:
Arecaceae; palm tree; somatic embryos; morphoanatomy; histochemical characterization

Authorship

Inaê Mariê de Araújo Silva-Cardoso

Universidade de Brasília, Departamento de Engenharia Florestal, Campus Universitário Darcy Ribeiro, 70.910-900, Brasília, DF, Brazil Universidade de BrasíliaBrazilBrasília, DF, BrazilUniversidade de Brasília, Departamento de Engenharia Florestal, Campus Universitário Darcy Ribeiro, 70.910-900, Brasília, DF, Brazil

Filipe Sathler Meira

Universidade de Brasília, Departamento de Biologia Celular, Campus Universitário Darcy Ribeiro, 70.910-900, Brasília, DF, Brazil Universidade de BrasíliaBrazilBrasília, DF, BrazilUniversidade de Brasília, Departamento de Biologia Celular, Campus Universitário Darcy Ribeiro, 70.910-900, Brasília, DF, Brazil

Ana Cristina Meneses Mendes Gomes

Embrapa Recursos Genéticos e Biotecnologia, PqEB, 70.770-917, Brasília, DF, Brazil. Embrapa Recursos Genéticos e BiotecnologiaBrazilBrasília, DF, BrazilEmbrapa Recursos Genéticos e Biotecnologia, PqEB, 70.770-917, Brasília, DF, Brazil.

Jonny Everson Scherwinski-Pereira ^** E-mail: jonny.pereira@embrapa.br

https://orcid.org/0000-0001-6271-332X

* E-mail: jonny.pereira@embrapa.br

SCIMAGO INSTITUTIONS RANKINGS

Figures

Figures (3)

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Figure 1
Morphoanatomical analysis of different responses of immature inflorescences of Syagrus oleracea submitted to somatic embryogenesis. A: Explant before inoculation. B: Swollen inflorescence after 30 days of culture. C: Inflorescence with discrete oxidation and callus initiation after 90 days of culture. D: Nodular and yellowish embryogenic callus after 270 days of culture. E: Anatomical segment of the callus mentioned in D; observe meristem and parenchyma zones. F: Starch grains (arrow) in the layer adjacent to the meristem zone evidenced by Periodic Acid Schiff - PAS and Lugol (side inset in detail). G: Embryogenic callus without definite shape and somatic embryos (arrow) after 270 days of culture. H: Anatomical segment of embryogenic callus; note the peripheral cells in intense cell division (lateral square). I: Transition of embryogenic callus into globular-like somatic embryo; observe protoderm in formation (arrow, side inset in detail) and starch grains (arrow, side inset in detail) in the layer adjacent to the meristem zone. Abbreviations: (ec) embryogenic callus; (inf) inflorescence; (mz) meristem zone; (pz) peripheral zone. Bars = A-D: 1mm; G: 2 mm; E, H, I: 200 µM and F: 50 µM.

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Figure 2
Morphological evolution of primary callus into plantlets of Syagrus oleracea from immature inflorescence in a medium with Picloram. A-D: Development of somatic embryos at 30, 60, 120 and 150 days, respectively, in induction medium. E, F: Somatic embryos in regeneration medium; note the presence of trichomes (*), green appearance and fusion. G: Fused plantlets. Bars = A: 1 mm and B-G: 2 mm.

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Figure 3
Morphoanatomical analysis of somatic embryos developed from embryogenic calluses of immature inflorescence, and anatomical and ultrastructural analysis of zygotic embryos of Syagrus oleracea. A: Somatic embryos. B: Surface of somatic embryo by Scanning Electron Microscopy. C: Somatic embryos with protoderm and procambium. D: Detail of the protoderm and ground meristem. E: Detail of the procambium. F: Ground meristem of zygotic embryo. G: Protein bodies stained with Xylidine Ponceau in cells of the ground meristem of zygotic embryo. H: Cell ultrastructure of the ground meristem of zygotic embryo; note conspicuous protein bodies and abundant lipid bodies. I, J: H in higher magnification. Abbreviation: (pb) protein body; (lb) lipid body; (se) somatic embryo; (ve) vessel element; (inf) inflorescence; (gm) ground meristem; (pc) procambium; (pt) protoderm. Bars = A: 2 mm; B, C: 500 µM; D, E: 50 µM; F, G: 20 µM; H: 2 µM and I, J: 0.5 µM.

Figure 1 Morphoanatomical analysis of different responses of immature inflorescences of Syagrus oleracea submitted to somatic embryogenesis. A: Explant before inoculation. B: Swollen inflorescence after 30 days of culture. C: Inflorescence with discrete oxidation and callus initiation after 90 days of culture. D: Nodular and yellowish embryogenic callus after 270 days of culture. E: Anatomical segment of the callus mentioned in D; observe meristem and parenchyma zones. F: Starch grains (arrow) in the layer adjacent to the meristem zone evidenced by Periodic Acid Schiff - PAS and Lugol (side inset in detail). G: Embryogenic callus without definite shape and somatic embryos (arrow) after 270 days of culture. H: Anatomical segment of embryogenic callus; note the peripheral cells in intense cell division (lateral square). I: Transition of embryogenic callus into globular-like somatic embryo; observe protoderm in formation (arrow, side inset in detail) and starch grains (arrow, side inset in detail) in the layer adjacent to the meristem zone. Abbreviations: (ec) embryogenic callus; (inf) inflorescence; (mz) meristem zone; (pz) peripheral zone. Bars = A-D: 1mm; G: 2 mm; E, H, I: 200 µM and F: 50 µM.

Figure 2 Morphological evolution of primary callus into plantlets of Syagrus oleracea from immature inflorescence in a medium with Picloram. A-D: Development of somatic embryos at 30, 60, 120 and 150 days, respectively, in induction medium. E, F: Somatic embryos in regeneration medium; note the presence of trichomes (*), green appearance and fusion. G: Fused plantlets. Bars = A: 1 mm and B-G: 2 mm.

Figure 3 Morphoanatomical analysis of somatic embryos developed from embryogenic calluses of immature inflorescence, and anatomical and ultrastructural analysis of zygotic embryos of Syagrus oleracea. A: Somatic embryos. B: Surface of somatic embryo by Scanning Electron Microscopy. C: Somatic embryos with protoderm and procambium. D: Detail of the protoderm and ground meristem. E: Detail of the procambium. F: Ground meristem of zygotic embryo. G: Protein bodies stained with Xylidine Ponceau in cells of the ground meristem of zygotic embryo. H: Cell ultrastructure of the ground meristem of zygotic embryo; note conspicuous protein bodies and abundant lipid bodies. I, J: H in higher magnification. Abbreviation: (pb) protein body; (lb) lipid body; (se) somatic embryo; (ve) vessel element; (inf) inflorescence; (gm) ground meristem; (pc) procambium; (pt) protoderm. Bars = A: 2 mm; B, C: 500 µM; D, E: 50 µM; F, G: 20 µM; H: 2 µM and I, J: 0.5 µM.

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Anatomical and histochemical studies of the somatic embryogenesis of <i>Syagrus oleracea</i> from immature inflorescences

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[1] * E-mail: jonny.pereira@embrapa.br