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| Da Silva,Rafael Fernandez; Menéndez-Yuffá,Andrea. |
| Different electroporation conditions were evaluated, toward the goal of transformation of Coffea arabica cv. Catimor. The tissues assayed were: embryogenic calli, leaf sections from in vitro plants, and somatic embryos in globular and torpedo stage obtained from cell suspensions. The effect of 1 or 24-hour pretreatment with an enzymatic solution (2% cellulase, 1% macerozyme) and electric field strength (375, 625, 875 V/cm) was evaluated. In all the experiments the tissues were incubated in ASP buffer (potassium aspartate) during three hours, and then one hour with plasmid DNA (pCambia3201, containing gus and bar genes) at room temperature. The electroporation was performed at a capacitance of 900 μF. The effect of the parameters evaluated was... |
| Tipo: Journal article |
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| Ano: 2003 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582003000100006 |
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| Menéndez-Yuffá,Andrea; Fernandez Da Silva,Rafael; Rios,Liliana; Xena de Enrech,Nereida. |
| Dividing cells of leaves used as sources of explants from coffee plants (Coffea arabica cv. 'Catimor') and those of their derived calli were analyzed for mitotic aberrations. The studied tissues were prepared by squashing and stained with carbolfuchsin. A total of 1551 leaf and 4568 callus cells were surveyed. The majority (79 %) of leaf and calli (75 %) cells showed normal mitosis, however, cells with mitotic abnormalities were also found in both tissues. These included: polyploids, aneuploids, sticky chromosomes, double prophases and lagging chromosomes. Additionally, interphase cells with micronuclei or binucleated were also observed. The frequencies of these abnormalities were statistically different in calli and leaves. Calli showed a few other... |
| Tipo: Journal article |
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| Ano: 2000 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582000000200008 |
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| Fernandez-Da Silva,Rafael; Menéndez-Yuffá,Andrea. |
| Two methods were evaluated in order to assess the viability of protoplasts and cell suspensions of Coffea arabica cv. Catimor used in a protocol of transformation by electroporation. One method consisted of staining with 1 % Evans blue and the other staining with 1% 3-[4,5-dimethyltiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). When Evans blue was applied to viable cells and protoplasts they either did not stain or acquired a faint blue colour. However, it was difficult to distinguish the non-viable cells in the wide spectrum of clear blue tonalities. In contrast, with the MTT assay only the viable cells and protoplasts reduced this salt to the red coloured formazan; viable and non-viable cells were distinguished more clearly with MTT than with Evans... |
| Tipo: Journal article |
Palavras-chave: Cell suspensions; Cell viability; Coffee; Evans blue; MTT; Protoplasts. |
| Ano: 2006 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500016 |
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