Somatic embryogenesis from immature and mature zygotic embryos of the acai palm (Euterpe oleracea): Induction of embryogenic cultures, morphoanatomy and its morphological characteristics

A somatic embryogenesis protocol for the acai palm (Euterpe oleracea Mart) was developed, based on mature and immature zygotic embryos, to define morphoanatomically the process's different stages and to analyze the homogeneity of nuclear DNA content by flow cytometry from calli, somatic embryos and regenerated plants. Auxin picloram (4-amino-3,5,6-trichloropicolinic acid) was tested to induce embryo genic calli at 225 and 450 mu M concentrations, coupled to the physiological maturing stages of zygotic embryos (mature and immature). Murashige and Skoog (MS) medium with 30 gl(-1) sucrose, 2.5 gl(-1) Phytagel, 2.5 gl(-1) activated charcoal and 0.5 gl(-1) L-glutamine was employed for callus induction. Embryo genic calli with somatic embryos in initial differentiation were transferred to a culture medium with 12.3 mu M of 2iP and 0.6 mu M of NAA and 300 mg l(-1) of activated charcoal for differentiating and maturing somatic embryos. Plant regeneration occurred in a medium with 1.0 mu M BAP (N-6-benzylaminopurine) and 0.5 mu M GA(3) (gibberellic acid). The formation of embryogenic calli in all treatments was observed in the induction medium, regardless of the stage of development of the zygotic embryo. Picloram at 450 mu M concentration provided the best results in forming embryogenic calli (84.7%). In the differentiating and maturing stage, 100% of the explants that had an embryogenic callus formation resulted in somatic embryos. The largest rate of plant regeneration (58.7%) was noted in treatment with an induction medium of 450 mu M of picloram and somatic embryos obtained from immature zygotic embryos. Morphoanatomical analyses evidenced that induction of somatic embryogenesis reflected stages characteristic of the indirect kind. Regenerated plants showed normal development, with growth of roots and aerial part. Calli, somatic embryos and plants, analyzed by flow cytometry, revealed no significant differences in the estimated rates of DNA content. (C) 2016 Elsevier B.V. All rights reserved.