simple metaphase chromosome preparation from meristematic root tip cells of wheat for karyotyping or in situ hybridization

Abstract

In this study, a simple and efficient metaphase chromosome preparation method from wheat was described which can also be applied on most other plants. The act of not using hydrolyzing chemicals such as hydrochloric acid in this method made the samples suitable for C-banding and in situ
hybridization as well. Squeezing out the meristematic cells from the root tip region on to the surface of the slide is the main step of the described method, leading to monolayer and clean preparations. In order to arrest cells in metaphase, colchicine, -monobromonaphthalene, ice cold water, colchicine-ice cold water and -monobromonaphthalene-ice cold water were compared to each other. Ice water pretreatment resulted in the highest metaphase index (up to 20% in some cultivars). Some preparations were used for in situ hybridization. The described method can be reliably applied in the laboratory with only basic equipments.