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Title of Journal: Plant Mol Biol Rep

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Abbravation: Plant Molecular Biology Reporter

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Springer-Verlag

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DOI

10.1016/0168-9002(94)01321-7

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1572-9818

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Flow Karyotyping of Wheat Addition Line “T240” wit

Authors: Dongwei Guo Donghong Min Zhaoshi Xu Ming Chen Liancheng Li Muhammad Ashraf Abdul Ghafoor Youzhi Ma
Publish Date: 2012/08/04
Volume: 31, Issue: 2, Pages: 289-295
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Abstract

To construct a chromosomespecific DNA library of chromosome 6VS from Haynaldia villosa a wheat alien chromosome addition line T240 with a 6VS chromosome arm and its parental common wheat cv CA921 were used to optimize protocols for preparing chromosome suspension amenable to flow sorting of the 6VS chromosome Our results showed that root tips incubated sequentially in Hogland’s solution containing 125 mM hydroxyurea DNA synthesis inhibitor for 18 h a hydroxyureafree period for 2 h and 1 μM trifluralin for 4 h metaphase blocking reagent increased the metaphase index MI by up to 62  Many metaphase chromosomes were released to form a chromosome suspension when root tips fixed in 2  paraformaldehyde were treated in a homogenizer at 9500 rpm for 10–15 s Most of the released chromosomes had intact morphology The background solution of chromosome suspension was clear and relatively few of cell debris and chromosome clumps Univariate flow karyotypes were established with chromosome suspension flow through FACS Vantage 2000 SE flow cytometer The flow karyotype of CA921 consisted of four chromosome peaks whereas that for T240 had a fifth peak This fifth peak in T240 contained the telosome which was further confirmed to be 6VS through fluorescence in situ hybridization

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