An improved technique for obtaining well-spread metaphases from plants with numerous large chromosomes. (3rd April 2017)

• Record Type: Journal Article
• Authors: Rodríguez-Domínguez, JM; Ríos-Lara, LL; Tapia-Campos, E; Barba-Gonzalez, R
• Is Part Of: Biotechnic & histochemistry. Volume 92:Number 3(2017)
• Page Start: 159
• Page End: 166
• Publication Date: 2017-04-03
• Subjects: acetic acid -- Amaryllidaceae -- chromosomes -- condensation -- cytogenetics -- Hymenocallis howardii -- metaphase -- spreading -- Sprekelia formosissima; Histochemistry -- Periodicals; Stains and staining (Microscopy) -- Periodicals; Biotechnology -- Periodicals; 572
• DOI: 10.1080/10520295.2017.1288927
• Languages: English
• ISSNs: 1052-0295

Abstract

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.