Technique for chromosome observation in Coffea embryos

Growing embryos excised from developing fruits are soaked in one of the three solutions: saturated solution of paradichlorobenzene, for 6-8 hours, 0.002 M 8-oxyquinoline, for 6-8 hours, of 0.2 percent colchicine, for 3-4 hours. After this they are fixed in 1:1 ethanolacetic acid overnight and then hydrolyzed in a mixture of 7 parts of 2 percent orcein in 70 percent acetic acid and 3 parts of hidrochloric acid, for 3 hours at 40 percent C. Next they are placed in a drop of 1 percent of acetic orcein on a slide and covered with a cover slip, pressure is then applied to flatten the tissue and separate the cells. Well coloured prophases, prometaphases and polar view metaphases with shortened and scattered chromosomes were observed in embryos of several species of Coffea

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Bibliographic Details
Main Authors: 92745 Medina, D.M., 58522 Conagin, C.H.T.M., 87506 Longo, C.R.L.
Format: biblioteca
Published: 1971
Subjects:COFFEA, CROMOSOMAS, EMBRION,
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Summary:Growing embryos excised from developing fruits are soaked in one of the three solutions: saturated solution of paradichlorobenzene, for 6-8 hours, 0.002 M 8-oxyquinoline, for 6-8 hours, of 0.2 percent colchicine, for 3-4 hours. After this they are fixed in 1:1 ethanolacetic acid overnight and then hydrolyzed in a mixture of 7 parts of 2 percent orcein in 70 percent acetic acid and 3 parts of hidrochloric acid, for 3 hours at 40 percent C. Next they are placed in a drop of 1 percent of acetic orcein on a slide and covered with a cover slip, pressure is then applied to flatten the tissue and separate the cells. Well coloured prophases, prometaphases and polar view metaphases with shortened and scattered chromosomes were observed in embryos of several species of Coffea