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Developmental pathway of somatic embryogenesis from leaf tissues of macaw palm (Acrocomia aculeata) revealed by histological events

quinta-feira, setembro 05, 2019

Fig. 1. Anatomical cross section of leaf segments of macaw palm (Acrocomia aculeata) during the initial stages of somatic embryogenesis induction. A: Leaf explant. B: Explant at 30 days of cultivation with hypertrophied cells (arrows). C: Explant after 60 days with onset of callus formation in the region of the vascular bundle. D: Explant with callus formation at a more advanced stage (side inset with detail of the callus). E: Calli formed from vascular bundles of the peripheral region of the explants. F: Detail of the callus in the vascular bundle region. Abbreviations: (ab) abaxial epidermis (ad) adaxial epidermis, (pc) primary callus, (p) phloem, (vb) vascular bundle, (st) stomata, (hy) hypodermis, (mp) mesophyll, (x) xylem. Scales: A, B, D, E, F: 0.1 mm and C: 0.02 mm.
Authors: Filipe Sathler Meiraa, Zanderluce Gomes Luisb, Inaê Mariê de Araújo Silva-Cardosoa, Jonny Everson Scherwinski-Pereirac

Abstract: This paper, in an unprecedented way, describes the ontogenesis of calli and somatic embryos of macaw palm (Acrocomia aculeata) from young leaf tissues of adult plants, aiming to elucidate the events underlying the process and consequent optimization of the technique. Leaves were inoculated in a callus induction medium, composed of Y3 salts, 2.5 g L−1 activated charcoal and 450 μM Picloram. For multiplication of the calli, they were separated from the explants and inoculated in a medium with the same composition as the previous phase. Histological analyses were performed on samples of immature leaves with and without calli at 0, 3, 15, 30, 60, 90 and 120 days in an induction medium, and on samples of the different types of calli and somatic embryos obtained during multiplication and differentiation. 

For this, the samples were fixed in FAA, dehydrated in an alcohol series and infiltrated in resin. Anatomical sections obtained on a microtome were stained with Toluidine Blue for structural characterization. At 60 days, analyses revealed the formation of callogenic masses in the vicinity of smaller vascular bundles. This multiplication approach allowed for the proliferation of the primary cell mass that evolved into new lineages of calli, especially the embryogenic yellowish nodular lineage, which originated somatic embryos. Additional histochemical analyses showed starch accumulation only in the adjacencies of areas undergoing intense cell division. 

The morpho-anatomic characterization allows the rapid identification of potential calli for the formation of somatic embryos from leaf tissues in macaw palm and promotes future investigations of the participation of vascular tissues in the events that precede somatic embryogenesis.

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