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A simple
metaphase chromosome preparation from meristematic root tip
cells of wheat for karyotyping or in situ
hybridization
Ghader Mirzaghaderi
Agronomy Department, Faculty of Agriculture, University of
Kurdistan, Sanandaj, Iran. E-mail:
mirzaghaderi@gmail.com or
gh.mirzaghaderi@uok.ac.ir.
Tel: +98 871 6627721-3. Fax: +98 871 6620553.
Accepted 26 November, 2009 |
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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, α-monobromonaphthalene,
ice cold water, colchicine-ice cold water and α-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.
Key words:
Fluorescence in situ hybridization (FISH),
karyotyping, squashing method, wheat. |
