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Large-scale propagation of oil palm (Elaeis guineensis, Jacq.) is difficult due to its single apical meristem. Thus, obtaining plants is mainly through seed germination, and a long growing period is required before oil production is possible. An alternative to large-scale seedling production is indirect somatic embryogenesis. The aim of this study was to analyze the somatic embryogenesis process in oil palm (E. guineensis Jacq.) with amino acids and low concentrations of auxins. The Tenera hybrid was analyzed by cytochemical and ultrastructural methods and was used to regenerate oil palm plants. First, calli were induced in MS culture media supplemented with 2,4-D and picloram. Two types of calli were obtained, characterized by beige or translucent color. Beige calli had embryogenic characteristics, such as large nuclei with prominent nucleoli, and they were multiplied for 8 months in MM culture (half strength MS, 1 mg L 2,4-D, 2 mg L 2iP, 1 mg L IBA, 250 mg L citric acid, 10 mg L cysteine, 100 mg L inositol, 1 mg L thiamine, 1 mg L pyridoxine, 1 mg L nicotinic acid, 1 mg L glycine, 200 mg L malt extract, and 100 mg L casein hydrolysate). After multiplication, the MCB culture medium (half strength MS, supplemented with 0.25 mg L NAA, 2 mg L BAP, MM vitamins and 200 mg L malt extract, and 100 mg L casein hydrolysate) was the most efficient for embryo formation, showing meristematic centers with totipotent cells in histochemical analyses. The somatic embryos were developed and germinated in MG medium (half strength MS, 0.45 mg L IAA, 0.25 mg L BAP, and MM vitamins), transplanted into polyethylene tubes containing pine bark substrates, and acclimatized in a greenhouse, achieving a 97% survival rate. The use of picloram for callus induction and somatic embryogenesis is advantageous and multiplication in MM medium is an important step for increasing cell mass. The calli with light beige color and nodular structures have meristematic cells with dense cytoplasm and totipotential features that later give rise to protoderm, procambium, and ground meristem during the globular, cordiform, and torpedo embryogenesis phases. In MCB medium, the concentration of vitamins and amino acids are crucial for somatic embryogenesis.
Large-scale propagation of oil palm (Elaeis guineensis, Jacq.) is difficult due to its single apical meristem. Thus, obtaining plants is mainly through seed germination, and a long growing period is required before oil production is possible. An alternative to large-scale seedling production is indirect somatic embryogenesis. The aim of this study was to analyze the somatic embryogenesis process in oil palm (E. guineensis Jacq.) with amino acids and low concentrations of auxins. The Tenera hybrid was analyzed by cytochemical and ultrastructural methods and was used to regenerate oil palm plants. First, calli were induced in MS culture media supplemented with 2,4-D and picloram. Two types of calli were obtained, characterized by beige or translucent color. Beige calli had embryogenic characteristics, such as large nuclei with prominent nucleoli, and they were multiplied for 8 months in MM culture (half strength MS, 1 mg L 2,4-D, 2 mg L 2iP, 1 mg L IBA, 250 mg L citric acid, 10 mg L cysteine, 100 mg L inositol, 1 mg L thiamine, 1 mg L pyridoxine, 1 mg L nicotinic acid, 1 mg L glycine, 200 mg L malt extract, and 100 mg L casein hydrolysate). After multiplication, the MCB culture medium (half strength MS, supplemented with 0.25 mg L NAA, 2 mg L BAP, MM vitamins and 200 mg L malt extract, and 100 mg L casein hydrolysate) was the most efficient for embryo formation, showing meristematic centers with totipotent cells in histochemical analyses. The somatic embryos were developed and germinated in MG medium (half strength MS, 0.45 mg L IAA, 0.25 mg L BAP, and MM vitamins), transplanted into polyethylene tubes containing pine bark substrates, and acclimatized in a greenhouse, achieving a 97% survival rate. The use of picloram for callus induction and somatic embryogenesis is advantageous and multiplication in MM medium is an important step for increasing cell mass. The calli with light beige color and nodular structures have meristematic cells with dense cytoplasm and totipotential features that later give rise to protoderm, procambium, and ground meristem during the globular, cordiform, and torpedo embryogenesis phases. In MCB medium, the concentration of vitamins and amino acids are crucial for somatic embryogenesis.
Oil palm is economically important as a crop with high oil production. Indirect somatic embryogenesis in oil palm requires a long time for callus induction and plant formation, and it is important to study the embryogenic potential of calli and the mechanisms of somatic embryogenesis. The aim of this study was to test different growth regulators and spermine in induction of embryogenic calli in root explants of oil palm (Elaeis guineensis Jacq). Apex root explants of approximately 0.5 cm were isolated from plants cultivated in vitro and inoculated in Y3 culture medium in the following treatments: A - without growth regulators; B - 1 mg.L-1 picloram (4-amino-3,5,6-trichloro-2-pyridinecarboxylic acid); C - 1 mg.L-1 picloram and 2 mg.L-1 2ip (2-isopentenyladenine); D - 2 mg.L-1 2ip; E - 1 mg.L-1 picloram and 2 mg.L-1 BAP (6-benzylaminopurine); F - 2 mg.L-1 BAP; and G - 14.5 mg.L-1 spermine. After six months of culturing, the calli induced in the treatments were analyzed by light microscopy. The calli induced in the treatments with 1 mg.L-1picloram (B) and treatment with 14.5 mg.L-1spermine (G) exhibited embryogenic characteristics, small and isodiametric cells, forming agglomerates, besides a large amount of starch. Calli of the best treatment (Y3 com 1 mg.L-1 de picloram) were inoculated in Y3 culture medium without addition of growth regulators. After eight months, calli were once more analyzed under light microscopy. All the treatments showed callus formation, except for treatments D and A. Calli of treatment B exhibited cells with embryogenic characteristics that developed somatic embryos.
The objective of this work was to evaluate the induction of indirect organogenesis by concentrations of dichlorophenoxyacetic acid (2,4-D) and thidiazuron (TDZ) in pitaya (Hylocereus undatus) explants, using scanning electron microscopy and the flow cytometry technique. The treatments consisted of the concentrations of 0, 2.0, and 4.0 mg L-1 2,4-D and TDZ and of the combinations of these regulators. Percentages of callus coverage at 45 and 60 days were evaluated. The explants subjected to the treatments were analized by flow cytometry and scanning electron microscopy. All treatments induced endoreduplication, and there was no somaclonal variation. Under the combination of 2.0 mg L-1 TDZ and 4.0 mg L-1 2,4-D, calluses were formed in 95% of the explants, but were smaller than those produced with 2,4-D separately. The concentration of 2.0 mg L-1 TDZ induces the indirect organogenesis in pitaya explants, confirmed by the presence of conducting vessels through scanning electron microscopy.
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