An improved technique for obtaining well-spread metaphases from plants with numerous large chromosomes |
| |
Authors: | JM Rodríguez-Domínguez LL Ríos-Lara E Tapia-Campos R Barba-Gonzalez |
| |
Institution: | 1. Centro de Investigación y Asistencia en Tecnología y Dise?o del Estado de Jalisco, A.C. Unidad de Biotecnología Vegetal, Guadalajara, Jalisco, Méxicorbarba@ciatej.mx;3. Centro de Investigación y Asistencia en Tecnología y Dise?o del Estado de Jalisco, A.C. Unidad de Biotecnología Vegetal, Guadalajara, Jalisco, México |
| |
Abstract: | Preparations that contain well-spread metaphase chromosomes are critical for plant cytogenetic analyses including chromosome counts, banding procedures, in situ hybridization, karyotyping and construction of ideograms. Chromosome spreading is difficult for plants with large and numerous chromosomes. We report here a technique for obtaining cytoplasm-free, well-spread metaphases from two Amaryllidaceae species: Sprekelia formosissima (2n = 120) and Hymenocallis howardii (2n = 96). The technique has three main steps: 1) pretreatment to cause chromosome condensation, 2) dripping onto tilted slides coated with a thin layer of pure acetic acid and 3) application of steam and acetic acid to produce cytoplasmic hydrolysis, which spreads the chromosomes. |
| |
Keywords: | acetic acid Amaryllidaceae chromosomes condensation cytogenetics Hymenocallis howardii metaphase spreading Sprekelia formosissima |
|
|