Abstract
Chromosome painting (CP) refers to visualization of large chromosome regions, entire chromosome arms, or entire chromosomes via fluorescence in situ hybridization (FISH). For CP in plants, contigs of chromosome-specific bacterial artificial chromosomes (BAC) from the target species or from a closely related species (comparative chromosome painting, CCP) are typically applied as painting probes. Extended pachytene chromosomes provide the highest resolution of CP in plants. CP enables identification and tracing of particular chromosome regions and/or entire chromosomes throughout all meiotic stages as well as corresponding chromosome territories in premeiotic interphase nuclei. Meiotic pairing and structural chromosome rearrangements (typically inversions and translocations) can be identified by CP. Here, we describe step-by-step protocols of CP and CCP in plant species including chromosome preparation, BAC DNA labeling, and multicolor FISH (Fig.1).
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References
Schubert I, Fransz PF, Fuchs J, de Jong JH (2001) Chromosome painting in plants. Methods Cell Sci 23:57–69
Lysak MA, Fransz PF, Ali HBM, Schubert I (2001) Chromosome painting in Arabidopsis thaliana. Plant J 28:689–697
Lysak M, Fransz P, Schubert I (2006) Cytogenetic analyses of Arabidopsis. In: Salinas J, Sanchez-Serrano JJ (eds) Methods in molecular biology, vol 323, Arabidopsis protocols. Humana, Totowa, NJ, pp 173–186
Szinay D, Chang S-B, Khrustaleva L, Peters S, Schijlen E, Bai Y et al (2008) High-resolution chromosome mapping of BACs using multi-colour FISH and pooled-BAC FISH as a backbone for sequencing tomato chromosome 6. Plant J 56:627–637
Febrer M, Goicoechea JL, Wright J, McKenzie N, Song X, Lin J et al (2010) An integrated physical, genetic and cytogenetic map of Brachypodium distachyon, a model system for grass research. PLoS One 5:e13461
Lysak M, Berr A, Pecinka A, Schmidt R, McBreen K, Schubert I (2006) Mechanisms of chromosome number reduction in Arabidopsis thaliana and related Brassicaceae species. Proc Natl Acad Sci USA 103:5224–5229
Ziolkowski PA, Kaczmarek M, Babula D, Sadowski J (2006) Genome evolution in Arabidopsis/Brassica: conservation and divergence of ancient rearranged segments and their breakpoints. Plant J 47:63–74
Mandáková T, Joly S, Krzywinski M, Mummenhoff K, Lysak MA (2010) Fast diploidization in close mesopolyploid relatives of Arabidopsis. Plant Cell 22:2277–2290
Lysak MA, Mandáková T, Lacombe E (2010) Reciprocal and multi-species chromosome BAC painting in crucifers (Brassicaceae). Cytogenet Genome Res 129:184–189
Henegariu O, Bray-Ward P, Ward DC (2000) Custom fluorescent nucleotide synthesis as an alternative method for nucleic acid labeling. Nat Biotechnol 18:345–348
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Kocsis E, Trus BL, Steer CJ, Bisher ME, Steven AC (1991) Image averaging of flexible fibrous macromolecules: the clathrin triskelion has an elastic proximal segment. J Struct Biol 107:6–14
Acknowledgments
We thank Dr A. Pecinka for providing seeds of the Sha × Col hybrid. This work was supported by grants IAA601630902 and P501/10/1014 from the Grant Agency of the Czech Academy of Science and the Czech Science Foundation (GA CR), respectively. German Science Foundation (DFG) and Alexander v. Humboldt Foundation are acknowledged for supporting our research (2003–2009).
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Lysak, M.A., Mandáková, T. (2013). Analysis of Plant Meiotic Chromosomes by Chromosome Painting. In: Pawlowski, W., Grelon, M., Armstrong, S. (eds) Plant Meiosis. Methods in Molecular Biology, vol 990. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-333-6_2
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DOI: https://doi.org/10.1007/978-1-62703-333-6_2
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