Establishment of protocol for decontamination of leaf explants of Platonia insignis Mart.
The objective of this work was the establishment of an efficient protocol for surface sterilization of leaf explants aiming at its introduction in callogenesis and somatic embryogenesis studies. The leaves were washed with distilled water and a detergent agent. In a laminar flow hood they were immersed in alcohol 70% (v/v) for 1 minute and then immersed in calcium hypochlorite solutions 0.25, 0.5, 0.75, 1.0, 1.25, 2.5, 5.0 or 10.0% (w/v) during 30 minutes, rinsed three times in distilled water, and cut in 1cm2 fragments, which were inoculated in MS medium. Half of the explants were inoculated with the abaxial epidermis in contact with the medium and half with the adaxial epidermis facedown. Each experimental unit was represented by 10 explants. The cultures were kept in the dark at 24±2ºC. Seven days after inoculation the percentages of contamination and necrosis of the explants were evaluated. The most efficient were the concentrations of 0.25, 0.5 and 0.75% calcium hypochlorite, which resulted in total sterilization of the explants. Treatments with 1.0, 1.25, 2.5, 5.0 and 10.0% calcium hypochlorite resulted in high level of necrosis (100%), independent of the side in contact with the medium. All the abaxial side facedown explants kept green and the adaxial ones became necrotic. It is recommendable 0.25% calcium hypochlorite in the surface sterilization and the abaxial epidermis in contact with the culture medium.
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Artigo de periódico biblioteca |
| Language: | English eng |
| Published: |
2016-01-14
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| Subjects: | Cultura de tecidos vegetais, Hipoclorito de cálcio, Estabelecimento in vitro, Descontaminação de explantes, Plant tissue culture, In vitro establishment, Explant decontamination., Platonia Insignis., calcium hypochlorite., |
| Online Access: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1033914 |
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| Summary: | The objective of this work was the establishment of an efficient protocol for surface sterilization of leaf explants aiming at its introduction in callogenesis and somatic embryogenesis studies. The leaves were washed with distilled water and a detergent agent. In a laminar flow hood they were immersed in alcohol 70% (v/v) for 1 minute and then immersed in calcium hypochlorite solutions 0.25, 0.5, 0.75, 1.0, 1.25, 2.5, 5.0 or 10.0% (w/v) during 30 minutes, rinsed three times in distilled water, and cut in 1cm2 fragments, which were inoculated in MS medium. Half of the explants were inoculated with the abaxial epidermis in contact with the medium and half with the adaxial epidermis facedown. Each experimental unit was represented by 10 explants. The cultures were kept in the dark at 24±2ºC. Seven days after inoculation the percentages of contamination and necrosis of the explants were evaluated. The most efficient were the concentrations of 0.25, 0.5 and 0.75% calcium hypochlorite, which resulted in total sterilization of the explants. Treatments with 1.0, 1.25, 2.5, 5.0 and 10.0% calcium hypochlorite resulted in high level of necrosis (100%), independent of the side in contact with the medium. All the abaxial side facedown explants kept green and the adaxial ones became necrotic. It is recommendable 0.25% calcium hypochlorite in the surface sterilization and the abaxial epidermis in contact with the culture medium. |
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