Abstract
In situ hybridization is a fundamental method in modern plant molecular cytogenetics research. It cannot be overstressed that good in situ hybridization results begin with good cytological preparations. Well-spread chromosome preparations with low amounts of cytoplasm give the best hybridization signals. Here, we present different protocols for the preparation of mitotic and meiotic plant chromosomes as well as tissue sections suitable for the detection of low-/single-copy sequences in a range of plant species. In addition a method suitable for the detection of single-copy probes is provided.
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References
Lavania UC (1998) Fluorescence in situ hybridization in genome, chromosome and gene identification in plants. Curr Sci India 74:126–133
Speicher MR, Carter NP (2005) The new cytogenetics: blurring the boundaries with molecular biology. Nat Rev Genet 6:782–792
Brady T, Clutter ME (1972) Cytolocalization of ribosomal cistrons in plant polytene chromosomes. J Cell Biol 53:827–832
Timmis JN, Deumling B, Ingle J (1975) Localisation of satellite DNA sequences in nuclei and chromosomes of two plants. Nature 257:152–155
Wimber D, Duffey P, Steffensen DM et al (1974) Localization of the 5S RNA genes in Zea mays by RNA-DNA hybridization in situ. Chromosoma 47:353–360
Rayburn AL, Gill BS (1985) Use of biotin-labeled probes to map specific DNA-sequences on wheat chromosomes. J Heredity 76:78–81
Schwarzacher T, Leitch AR, Bennett MD et al (1989) In situ localization of parental genomes in a wide hybrid. Ann Bot 64:315–324
Yamamoto M, Mukai Y (1989) Application of fluorescence in situ hybridisation to molecular cytogenentics of wheat. Wheat Inf Serv 69:30
Jiang JM, Gill BS (2006) Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research. Genome 49:1057–1068
Jiang JM, Gill BS (1994) Nonisotopic in situ hybridization and plant genome mapping – the first 10 years. Genome 37:717–725
Silva GG, Souza MM (2013) Genomic in situ hybridization in plants. Genet Mol Res 12:2953–2965
Lysak MA, Fransz PF, Ali HB et al (2001) Chromosome painting in Arabidopsis thaliana. Plant J 28:689–697
Gill B, Friebe B (1998) Plant cytogenetics at the dawn of the 21st century. Curr Opin Plant Biol 1:109–115
Kato A, Albert PS, Vega JM et al (2006) Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation. Biotech Histochem 81:71–78
Danilova TV, Friebe B, Gill BS (2014) Development of a wheat single gene FISH map for analyzing homoeologous relationship and chromosomal rearrangements within the Triticeae. Theor Appl Genet 127:715–730
Aliyeva-Schnorr L, Beier S, Karafiátová M et al (2015) Cytogenetic mapping with centromeric BAC contigs shows that this recombination-poor region comprises more than half of barley chromosome 3H. Plant J 84:385–394
Valárik M, Bartos J, Kovárová P et al (2004) High-resolution FISH on super-stretched flow-sorted plant chromosomes. Plant J 37:940–950
Fransz PF, Alonso-Blanco C, Liharska TB et al (1996) High-resolution physical mapping in Arabidopsis thaliana and tomato by fluorescence in situ hybridization to extended DNA fibres. Plant J 9:421–430
Aliyeva-Schnorr L, Ma L, Houben A (2015) A fast air-dry dropping chromosome preparation method suitable for FISH in plants. J Vis Exp 106:e53470
Sanchez Moran E, Armstrong SJ, Santos JL (2001) Chiasma formation in Arabidopsis thaliana accession Wassileskija and in two meiotic mutants. Chromosome Res 9:121–128
Steedman HF (1957) Polyester wax; a new ribboning embedding medium for histology. Nature 179:1345
Braszewska-Zalewska AJ, Wolny EA, Smialek L et al (2013) Tissue-specific epigenetic modifications in root apical meristem cells of Hordeum vulgare. PLoS One 8:e69204
Muravenko OV, Amosova AV, Samatadze TE et al (2003) 9-Aminoacridine: an efficient reagent to improve human and plant chromosome banding patterns and to standardize chromosome image analysis. Cytometry A 51:52–57
Armstrong SJ, Jones GH (2003) Meiotic cytology and chromosome behaviour in wild-type Arabidopsis thaliana. J Exp Bot 54:1–10
Acknowledgments
The authors thank the members of the laboratories for a helpful discussion. C.M. is the recipient of a Marie Curie Initial Training Network fellowship (FP7-PEOPLE-2013-ITN, CHIP-ET).
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Badaeva, E.D., Ruban, A.S., Aliyeva-Schnorr, L., Municio, C., Hesse, S., Houben, A. (2017). In Situ Hybridization to Plant Chromosomes. In: Liehr, T. (eds) Fluorescence In Situ Hybridization (FISH). Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-52959-1_49
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DOI: https://doi.org/10.1007/978-3-662-52959-1_49
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