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http://ciatej.repositorioinstitucional.mx/jspui/handle/1023/786| Isolation and regeneration of protoplasts from leaf explants of Rhyncholaelia digbyana | |
| LUIS ALBERTO MOTA NARVAEZ GILBERT JOSE HERRERA COOL Teresa del Rosario Ayora Talavera NANCY SANTANA BUZZY Guadalupe Lopez Puc | |
| Acceso Abierto | |
| Atribución-NoComercial-SinDerivadas | |
| DOI: 10.5897/AJB2018.16540 | |
| "A protocol for the isolation and regeneration of protoplasts from leaf explant of Rhyncholaelia digbyana is presented. The protoplasts were isolated using hemicellulase enzymes at 1.5, 2.25, and 3% (w/v), pectinase at 0.5 and 0.75% (w/v) and cellulase at 1 and 2% (w/v). Protoplast counting was carried out with a Neubauer camera and an optical microscope at 40X, and viability was determined with Evans blue dye at 0.025% (w/v). The protoplasts were cultivated following the standard plate method, using the K&M media with 0.06 M of saccharose, 2.3% of Gelrite and plant growth regulators. It produced a yield of 386250±1875 protoplasts/g of tissue using an enzymatic combination of 1.5% (w/v) of hemicellulase, 0.5% (w/v) of pectinase and 1% (w/v) of cellulase with an incubation time of 4 h. The colonies were observed after two months of culture and the highest number of colonies (1.66±0.50) was obtained when the protoplasts were cultured in Kao medium with 4.53 μM of 2,4-Dichlorophenoxyacetic acid (2,4-D), 0.912 μM of Zeatin and 3.10 μM of 6-Benzylaminopurine (BA). The nuclear DNA content estimated by flow cytometry for R. digbyana was 27.39±3.8 pg of DNA equivalent to 26.79×109 pb and the number of mitotic chromosomes counted was 2n=40". | |
| Academic Journals | |
| 2018-08 | |
| Artículo | |
| African Journal of Biotechnology Vol. 17(35), pp. 1082-1089, 29 August, 2018 | |
| Inglés | |
| Bibliotecarios Empresas Estudiantes Investigadores Maestros | |
| CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA | |
| Versión publicada | |
| publishedVersion - Versión publicada | |
| Aparece en las colecciones: | ARTICULOS |
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